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Table of Contents - Current issue
Apr-Jun 2022
Volume 18 | Issue 78
Page Nos. 247-517
Online since Thursday, July 7, 2022
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ORIGINAL ARTICLES
Tinospora cordifolia
ameliorates behavioral deficits in conditioned fear and single prolonged stress–induced preclinical PTSD model in mice by modulating translocator protein (18kDa, TSPO)
p. 247
Gaurav Ranjan, Priyashree Sunita, Pritha Bose, Sugato Banerjee, Shakti P Pattanayak
DOI
:10.4103/pm.pm_89_22
Background:
Post-traumatic stress disorder (PTSD) is a severe chronic psychiatric condition for which currently there is no specific therapy. Translocator protein (18 kDa; TSPO), a critical therapeutic target for treating PTSD and other neurological deficits regulates allopregnanolone biogenesis. Allopregnanolone potently mediates allosteric modulation of GABA
A
which subsequently coordinates emotional behavior. The well-known ayurvedic plant
Tinospora cordifolia
is rich in therapeutically active phytoconstituents which contribute toward its diversified efficacy against neurological disorders like anxiety, depression, etc.
Materials and Methods:
In the present study, we explored the potency of
Tinospora cordifolia
in modulating TSPO to improve PTSD symptomatology in preclinical mice model of combined model of conditioned fear (electric foot shock) and single prolonged stress to induce PTSD. A series of behavioral assessments, histopathological investigations, ELISA and Western blot analysis were conducted to decipher a potential molecular anti-PTSD mechanism of ethanolic extract of
Tinospora cordifolia
extract TnCE.
Results:
Following the treatment protocol, TnCE revealed prominent anxiolytic and antidepressant activity in PTSD-afflicted mice with simultaneous improvement of social behavior and attenuated context memory. Interestingly, the positive impact of TnCE was completely abolished by TSPO selective antagonist PK11195. Moreover, consistent upregulation of TSPO expression with a marked escalation in APG and GABA levels indicated a TSPO-dependent mechanism underlying the pharmacotherapeutic efficacy of TnCE in the PTSD model.
Conclusion:
Thus, this multi-faceted beneficial TnCE may offer a novel therapeutic entity for PTSD treatment by modulating TSPO mediated allopregnanolone biogenesis.
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Commigileadin A: A new triterpenoid from
Commiphora gileadensis
aerial parts
p. 256
Hossam M Abdallah, Gamal A Mohamed, Sabrin R M. Ibrahim, Abdulrahman E Koshak, Ibrahim Alnashri, Abdullah Alghamdi, Abdulghani Aljohani, Abdulrahman Khairy
DOI
:10.4103/pm.pm_118_21
Background:
Genus
Commiphora
(Burseraceae) includes about 150 species of aromatic shrubs located in Africa, India, and the Arabian region.
C. gileadensis
L. has been used for treating several ailments such as constipation, urinary retention, headache, jaundice, joint pain, liver and stomach diseases, and inflammatory disorders.
Objectives:
In this work, a phytochemical investigation of
C. gileadensis
aerial parts was carried out. Moreover, the antimicrobial potential of the new metabolite was assessed.
Materials and Methods:
The aerial parts were extracted at room temperature with methanol (MeOH). The MeOH extract was subjected to various chromatographic techniques to separate the bio-constituents. Their structures were exhaustively specified by utilizing diversified spectroscopic data and comparing them with the literature. The antimicrobial potential of the new compound
1
was assessed toward
Bacillus cereus, Escherichia coli, Staphylococcus aureus,
and
Clostridium albicans
.
Results:
A new friedelan triterpenoid, commigileadin A [3,11-dioxo-(D: A)-friedo-olean-27-al] (
1
) and four known metabolites, namely, Stigmasterol (
2
), naringenin (
3
), naringenin-4-methyl ether (
4
), and kaempferol (
5
) were obtained. Compound
1
possessed moderate activity toward
B. cereus
(inhibition zone diameter (IZD): 12.6 mm and minimum inhibitory concentration [MIC]: 8.9 μg/mL) in comparison to ciprofloxacin (IZD: 21.1 mm and MIC: 2.5 μg/mL, respectively).
Conclusion:
This work reported the characterization of a new triterpenoid (
1
) and four known constituents (
2-5
). Compound
1
possessed a moderate activity toward
B. cereus.
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Anti-inflammatory and antiallergic activity of arbutin against ovalbumin-induced allergic rhinitis in mice through the modulation of inflammatory responses
p. 261
Tao Zhang, Xiaopeng Sun, Ruiping Fang
DOI
:10.4103/pm.pm_14_21
Background:
Allergic rhinitis (AR) is the prevalent inflammatory disease in the airway due to allergic reactions in response to numerous allergens. AR is considered as a type-I allergic condition and proceeded as early- and late-phase hypersensitivity.
Objectives:
In the existing work, we scheduled to discover the anti-inflammatory potential of arbutin against the ovalbumin (OVA)-provoked AR in mice through modulation of inflammation.
Materials and Methods:
The AR was triggered to the BALB/c mice by injecting 500 μL of OVA sensitization solution and then treated with the arbutin (25 and 50 mg/kg, respectively). Dexamethasone was used as standard. The occurrences of sneezing and nasal rubbing were detected within 15 min after the last OVA challenge. The status of OVA-specific immunoglobulin E (IgE), histamine, and malondialdehyde (MDA) was scrutinized by using assay kits. The status of NF-κB/IκBα and STAT-3 signaling molecules and its related cytokines was considered by using assay kits. The histological scores were evaluated by the histological alterations.
Results:
The arbutin supplementation lessened the incidence of nasal rubbings and sneezing, OVA-specific IgE and histamine, and MDA status. The arbutin-administered AR mice established the appreciable reduction in the NF-κBp65, phosphorylated IκBα, interleukin (IL)-1β, and tumor necrosis factor-α levels, also improved the IκBα status in the AR mice. Arbutin reduced the STAT-3, phosphorylated STAT-3, RORc, IL-17A, IL-5, and IL-6 levels and elevated the IL-10, IL-12, and IFN-γ status. Arbutin supplementation to the AR mice exhibited the considerable amelioration of the histological scores of OVA-provoked AR mice.
Conclusion:
Our results established that the arbutin treatment effectively ameliorated the OVA-provoked AR in mice through the modulation of inflammation, and it could be auspicious therapeutic agent to treat the RA.
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Goniothalamin-mediated amelioration of doxorubicin-induced myocardial damage and regulation of nuclear factor-κB/HO-1/NQO-1 signaling biomarkers in cardiotoxic rats
p. 267
Chong Liu, Qiuju Wang
DOI
:10.4103/pm.pm_180_21
Background:
Chronic use of doxorubicin (DOX) as an anticancer and antineoplastic agent has a chief jeopardy of cardiotoxicity. About 10% of the treated population has logged to cause cardiac damage.
Objectives:
This study principally engrossed on inspecting the effective cardioprotective activity of goniothalamin (GTN) against DOX-induced cardiotoxic rats.
Materials and Methods:
Group I – control, Group II – inducer (DOX) alone (2.5 mg/kg body weight [BW]) given on alternate days, Group III – DOX + GTN (2.5 mg/kg BW + 200 mg/kg BW), and Group IV – GTN alone (200 mg/kg BW). First, it employed its protective effects over the isolated cardiac tissues in which the status of HO-1 and NQO-1 were upregulated.
Results:
GTN administered with DOX induced rats were showed the increased the status of antioxidant levels, elevation of reactive oxygen species, which is also reduced the inflammatory and stress markers contributing to its cardio protective activity. Furthermore, GTN also downregulated the mRNA expression status of inflammatory markers and HO-1, NAD (P) H, and NQO-1 in DOX-induced rats, thereby weakening the cardiac damage.
Conclusion:
GTN is an effective protective agent against DOX-induced cardiotoxicity in rats.
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Characterization of PLGA-PEG
Catharanthus roseus
nanoparticles and assessing its anticancer effects in her2-overexpressed breast cancer cells
p. 273
ZS Siti, Nor Hazwani Ahmad, Shahrul Hamid
DOI
:10.4103/pm.pm_338_21
Background:
Catharanthus roseus
(
CR
) shows promising anticancer activity. However, there is limited information on its polymeric formulation.
Objectives:
Therefore, current study aimed to characterize poly(lactic-co-glycolic acid (PLGA)
CR
nanoparticles and determine its effects on resistant human epidermal receptor 2 (HER2)-overexpressed breast cancer cells.
Methods:
PLGA-polyethylene glycol (PEG)
CR
nanoparticles were synthesized using the solvent displacement method and characterized using ultraviolet-visible spectroscopy (UV-VIS), Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering, transmission electron microscopy (TEM), zeta potential, encapsulation efficiency, and drug release experiments. Cytotoxicity was done using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Protein expression was done with a gel image analyzer. Cell morphological changes were viewed under phase-contrast microscope.
Results:
TEM images showed the nanoparticles were spherical and size less than 100 nm. FTIR results indicated encapsulation of
CR
based on the presence of 3327 cm
–1
, 1637 cm
–1
, and 1066 cm
–1
peaks. Encapsulation efficiency was >60% in both formulations. However, pluronic F68 PLGA-PEG CR nanoparticles showed a gradual release of
CR
compared with polyvinyl acetate (PVAc). The cytotoxicity assay showed that the half-maximal inhibitory concentration of the
CR
nanoparticles generated with F68 and PVAc was lower (42–58 μg/mL) on tamoxifen-resistant cells compared with parent cells (99–147 μg/mL). Further analysis using CR nanoparticles with F68 exhibited downregulation of HER2 expression and induced apoptotic features based on morphological changes.
Conclusion:
These findings suggest that PLGA-PEG nanoparticles could retain the cytotoxic effects of
CR
.
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Rehmanniae Radix
leaves stimulates ROS-induced apoptosis on human mammary cancer cells through suppressing PI3K/AKT/mTOR and GSK3β signaling pathway
p. 281
Chao Yuan, Peihua Sun, Xin Ge, Minling Xu
DOI
:10.4103/pm.pm_144_21
Background:
Mammary carcinoma is the most communal carcinoma of higher women mortality.
Rehmanniae Radix
(RR) a traditional Chinese medicine.
Objectives:
To examine the anti-proliferative and anti-apoptotic effect of RR plant extract on mammary cancer cells MCF-7 and its action on PI3K/AKT/mTOR and GSK3 β mechanisms.
Materials and Methods:
The anti-proliferative of plant extract of RR was calculated, and morphological changes were measured by 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide assay. ROS (dichloro-dihydro fluorescein diacetate), MMP (Rh-123), apoptosis by Propidium Iodide, 4' 6-diamidino-2-phenylindole, and Acridine Orange/Ethidium Bromide staining assays assessed cellular cell death in MCF-7 cells. Western blotting analyses exhibited both phosphorylated forms compared with total PI3K/AKT/mTOR and GSK3 β were recognized.
Results:
The inhibitory action of breast cancer cells that RR extracts designated in a concentration manner. The IC
50
value of RR extract on MCF-7 cells at 150 μg/mL concentration. Furthermore, staining assays seemed that RR extract-treated at 150 and 200 μg/mL, ROS production and apoptosis were augmented, and MMP and cell development were meaningfully reduced compared to control cells. Western blot analysis severely reduced the expression of p-PI3K, p-AKT, p-mTOR, and p-GSK3 β, but all total was extremely expressed than control cells. Altogether, the initiation of programmed cell death and suppression of cell propagation by the action of RR extract through the PI3K/AKT/mTOR mechanism recommended the anti-proliferative action of RR.
Conclusion:
Our results gave an innovative place of anti-tumor effect of RR extract is a gifted therapeutic and preventive agent in mammary cancer cell treatment.
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Xylocarpus moluccensis
fruit fraction rescues cardiac hypertrophy by improving angiogenesis and regulating NF-κB-mediated inflammation
p. 286
Amit Manhas, Dipika Goyal, Bharti Biswas, Dipti Tripathi, Pragya Yadav, Abhinav Singh, Shri Krishna, Narender Tadigoppula, Madhu Dikshit, Kumaravelu Jagavelu
DOI
:10.4103/pm.pm_79_21
Background and Aim:
The present study was undertaken to investigate the potential of ethyl acetate fraction obtained from the fruits of
Xylocarpus moluccensis
alcoholic extract (CDR-267-F018) against cardiac hypertrophy in rats.
Experimental Procedure
:
Cardiac hypertrophy was achieved in Wistar rats through isoproterenol and treated either with propranolol or CDR-267-F018 for 14 days.
Results and Conclusion:
CDR-267-F018 treatment reduced isoproterenol induced cardiac hypertrophy as assessed by 2D-echocardiography and supported by reduction in
ANP, BNP, β-MHC, NPP-A
and increased expression of vascular endothelial growth factor receptor 1. CDR-267-F018 treatment tightly regulated inflammation by controlling the plasma proinflammatory cytokines tumor necrosis factor-α and IFN-γ, plasma EMP level and NF-κB, Akt and ERK. Further, CDR-267-F018 treatment reduced fibrosis by regulating
Col18a1, FGF-21
and
MMP2
. In total, CDR-267-F018 protected rat heart against isoproterenol induced cardiac hypertrophy by acting on inflammation, fibrosis and by improving angiogenesis.
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Cratoxylum formosum
leaf extracts inhibit growth, induce apoptosis, and decrease metastasis of hela human cervical cancer cells
p. 296
Benjaporn Buranrat
DOI
:10.4103/pm.pm_442_21
Background:
Cratoxylum formosum
(CF) is being used in Asian countries to treat disease and for eating purposes.
Objectives:
In the present study, the young leaf extract of CF was extracted by distilled water (DW) and 95% ethanol and examined on anti-cancer actions in cervical cancer cells.
Materials and Methods:
CF was determined the cytotoxic effects by sulforhodamine B, colony formation, cell cycle arrest, apoptosis, ROS formation, and mitochondrial function assay.
Results:
The data revealed that the Distilled water (DW) and ethanol (EtOH) extract had potent cytotoxicity against HeLa cells, and EtOH extract had more potency than DW extract. Inhibition of colony-forming ability was related with cell growth and at 250 mg/mL of CF was showed the highest activity. In addition, cancer cell distribution was stopped at the G0/G1 phase after incubating with 250 μg/mL of both extracts. The extracts potently induced cancer cell apoptosis properties in a dose-dependent manner. DW extract induced late apoptosis from 4.7%, 6.2%, 7.8%, and 8.0% (0, 50, 100, 250 μg/mL); however, EtOH extract induced early apoptosis from 3.4%, 5.7%, 6.1% and 16.0%, respectively. Consistent with apoptotic effects, EtOH extract reduced mitochondrial function and the higher effects were found at 250 μg/mL concentrations and related to the ROS production. The two CF extracts powerfully inhibited the migratory abilities of HeLa cells. The data indicated that EtOH of CF extract had more potency against HeLa cells than DW extract.
Conclusion:
CF extracts are potent against HeLa cells proliferation, induces apoptosis, and suppresses migration; use of this plant for cervical cancer treatment should be encouraged.
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Efficacy of bisacurone in bone fracture healing process: An experimental study on osteoporotic rats
p. 304
Minjie Li, Weiwei Feng, Wenping Zhang, Zhonghui Pang, Shaokun Pei, Lifeng Xia, Qiang Li, Huixiao Li
DOI
:10.4103/pm.pm_450_21
Background:
Osteoporosis (OP) is a common disorder resulting in bone fragility and fracture. Due to OP, bone mass and microarchitectural bone tissues are damaged, increasing the risk of fracture.
Objectives:
To determine the efficacy of bisacurone in healing fractured bones and its inflammatory response in ovariectomy (OVX)-induced OP in female rats.
Materials and Methods:
After inducing fracture in femur bone, bisacurone (25, 50, and 100 μg/kg) and alendronate (20 mg/kg) were orally administered to rats for 8 weeks. Subsequently, its fracture healing and anti-inflammatory potential were evaluated via assessment of various biochemical and molecular parameters.
Results:
Bisacurone therapy significantly (
P
< 0.05) increased the calcium content, serum calcium, and phosphorus in OVX-induced OP rats. It significantly (
P
< 0.05) reduced the serum alkaline phosphate (ALP); urine biochemical parameters such as urinary calcium, phosphorus, and creatinine; and inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Moreover, bisacurone was found to be significantly (
P
< 0.05) effective in downregulating the bone turnover markers such as osteocalcin, receptor activator of nuclear factor-kappa-γ ligand (RANKL), peroxisome proliferator-activated receptor gamma (PPAR-γ), and upregulating osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2), and AMP-activated protein kinase (AMPK) in OVX-induced OP rats.
Conclusion:
Bisacurone effectively healed the fracture and improved the bone quality in arthritic conditions by reducing inflammatory cytokines and altering the level of bone turnover markers. Therefore, bisacurone therapy should be considered for the management of fracture healing process in the arthritic conditions.
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Exploring cross-linked tragacanth as novel excipient-proof-of-concept
p. 314
Anroop B Nair, Santosh Fattepur, N Raghavendra Naveen, Prakash Goudanavar, Naga Prashant Koppuravuri, Buduru Gowthami, Mallikarjun Telsang, Riyaz Ali Osmani, Nagaraja Sreeharsha, Mohammed Habeebuddin
DOI
:10.4103/pm.pm_601_21
Background:
Tragacanth, a natural gum, is frequently used as stabilizer for colloidal systems and as a binder in tablets. Materials from natural sources are in increasing demand to solve the current global environmental problems arising from synthesis involving petroleum-based substances.
Objectives:
In this context, we improved functionality of tragacanth through crosslinking and extended its application for directly compressed fast dissolving systems. Fast dissolving formulations upon settling on the tongue disintegrate promptly and release the medicament, thus making it especially suitable for paediatrics, geriatrics, bedbound, or incapacitated patients.
Materials and Methods:
Cross-linked tragacanth (CLT) was explored as a potent disintegrant and compared with sodium starch glycolate and Crospovidone for its effect on compressibility and release of metoclopramide hydrochloride from tablets made by direct compression and sublimation method. Formulations made using CLT were optimized for swelling capacity, absorption efficiency, and moisture sorption capacity.
Results:
The most appropriate controls for linkage of tragacanth were 1:0.4 proportion of tragacanth: Epichlorohydrin, at 105°C temperature for 45 min of reaction. Prepared formulations showed desired disintegration and wetting time. Formulations made using camphor showed porosity because of sublimation and favored rapid disintegration. Based on the drug release study, it is confirmed that formulation with 4% CLT and 20% camphor prepared by sublimation process exhibited highest drug release, i. e. 99.23% within 15 min.
Conclusion:
This study demonstrates the novel applicability of tragacanth as an effective natural superdisintegrant after cross-linking and provides a sustainable alternative to synthetic superdisintegrants while formulating the fast-disintegrating tablets.
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Preparation of a nanoemulsified drug delivery system of astilbin using edible formulation components:
In vivo
evaluation of oxaliplatin-induced cirrhotic liver injury
p. 321
Yixing Wang, Xuping Zhang, Jin Zhou, Shilei Xu, Xiaodan Kou, Yuan Cheng
DOI
:10.4103/pm.pm_345_21
Background:
Astilbin is a flavonoid phytoconstituent extracted from Chinese herb Rhizoma Smilacis Glabrae, a common Chinese food ingredient.
Objectives:
The present study used astilbin as a model drug to develop food-based nanoemulsified formulations. These formulations were then tested for their possible role in countering Oxaliplatin (OXP) associated oxidative stress.
Materials and Methods:
Thirty six healthy male BALB/c mice of age 10–12 weeks and weight 25–30 g were included in the present study. The mice were administered OXP through intraperitoneal route in a dose of 8 mg/kg for a period of 4 days. The animals in the control group were administered a 10 ml/kg dose of 5% (w/v) glucose solution. For evaluating the hepatoprotective effect of developed formulations on OXP-induced liver injury, the animals from treatment groups were administered once daily dose of drug loaded formulations 60 min before OXP treatments (200 mg/kg equivalent of formulation, intraperitoneally) for 4 days. Finally, the blood and hepatic tissue samples were obtained for future analysis.
Results:
We successfully developed non-toxic nanoemulsion systems for astilbin, which comprised of different non-toxic food ingredients of astilbin and evaluated the potential reversing the OXP induced liver injury. The food-based nanoemulsions possessed attractive physical properties and exhibited sufficient stability profile when subjected to freeze-thaw cycles as well as 6-month storage at ambient and cold temperatures. Furthermore, excellent cumulative
in vitro
drug permeation was reported for the prepared formulations through dialysis membrane bags. We also observed the significant reduction in biomarkers of hepatic oxidative stress, caused by OXP chemotherapy in experimental mice, thereby indicating a marked reversal of liver injury.
Conclusion:
We concluded that nanoemulsified astilbin can be used as therapeutic agent to counter OXP-associated oxidative stress.
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Antitumor and immunomodulation of total flavonoids from the green peep of
Juglans regia
L. in mouse forestomach carcinoma gastric cancer-bearing mice
p. 328
Baozhong Wei, Hongxia Zhu, Kai Wang, Shuangming Liu, Yihui Dai, Danhong Liu, Fengguo Xia, Hua Sun
DOI
:10.4103/pm.pm_54_21
Background:
Gastric cancer is a malignant tumor of the digestive tract. It is the fifth most common cancer and the third leading cause of cancer death across the world. In this study, we investigated the antitumor and immunomodulatory effects of total flavonoids from the green peel of
Juglans regia
L.(JRFs) in Mouse Forestomach Carcinoma (MFC) gastric cancer-bearing mice.
Methods:
In this study, an MFC gastric cancer-bearing mouse model was established. The killing activity of natural killer (NK) cells, the proliferation of lymphocytes, and the phagocytosis of macrophages of mice were observed by lactate dehydrogenase release assay and MTT assay. The levels of tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), and IL-6 in the serum of mice were detected by enzyme-linked immunosorbent assay, and the mRNA and protein expressions of Toll-Like Receptor 4 (TLR4), MyD88, and nuclear factor kappaB (NF-κB) in the spleen tissue were detected by real-time polymerase chain reaction and Western blot analysis, respectively.
Results:
Compared with the model group, the tumor weight of mice in the high-, medium-, and low-dose JRF groups was significantly decreased, whereas the spleen index, the activity of NK cells, the proliferation of lymphocytes, and the phagocytosis of macrophages were significantly increased. There was no significant difference in the level of TNF-α, IL-2, and IL-6 in the serum of mice between the model group and the low-dose JRFs group, whereas the levels of TNF-α, IL-2, and IL-6 in the serum of mice in the high-and medium-dose JRF groups were significantly higher than those in the model group. The mRNA and protein expressions of TLR4, MyD88, and NF-κB in the spleen tissue of mice in the high-, medium-, and low-dose JRF groups were significantly lower than those in the model group.
Conclusion:
JRFs can inhibit the growth of MFC gastric cancer in mice and the blocked activation of TLR4/NF-κB signaling pathway may be the potential mechanism of the antitumor and immunomodulatory effects of JRFs in MFC gastric cancer-bearing mice.
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Oridonin Regulates NF-κB Signaling Pathway in Gestational Diabetes Suppression of SPARC and G6pase in C57BL/6J Mice
p. 334
Jinjin Qin, Bei Wang, Rui Li, Fang Wang, Yan An, Chenyuan Cao
DOI
:10.4103/pm.pm_146_21
Background:
In recent years, the morbidity of gestational diabetes (GD) has increased by around 5%–10%, and it now affects around 8%–21% of all births, with around 3%–5% of these women suffering from long-term diabetes after birth. Premature maturation, birth trauma, macrosomia, and respiratory problems are the common complications seen in pregnant women with GD. Lack of glucose tolerance is normally found in pregnant women with GD. Furthermore, GD during pregnancy can result in more complications in the long run, with a high risk of type-2 diabetes developing in subsequent generations.
Materials and Methods:
C57BL/KsJ mice were used to evaluate the efficacy of oridonin against GD. The animals were divided into four groups: normal mice with pregnancy, GD alone; GD + oridonin (25 mg/kg bw), and GD + oridonin (50 mg/kg bw). After 10 days of gestational period, the following parameters were evaluated: glucose and insulin tolerance in blood, body weight, lipid peroxidation products (thiobarbituric acid reactive substances (TBARS)), and antioxidant markers (superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) in liver tissues were analyzed. After 20 days of gestational period, the following parameters were measured in the liver tissues by using ELISA kits: elevation of secreted protein acidic and rich in cysteine (SPARC) and pro-inflammatory cytokines.
Results:
In GD-induced mice, oridonin partially corrected glucose and insulin tolerance and maintained ideal body weight. Moreover, oridonin elevated the levels of SOD, CAT, and GSH and reduced the levels of TBARS in GD mice. Finally, oridonin downregulated the expression of SPARC and nuclear factor kappa B (NF-κB).
Conclusion:
In summary, oridonin showed an anti-inflammatory antioxidant effect and prevented GD in pregnant mice.
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Potential role of
Tribulus terrestris
fruit extract in inhibition of advanced glycation end products
p. 341
Dinesh Kumar, Nidhi Shah, Khalid M Al-Anazi, Mohammad Abul Farah, M Ajmal Ali, Joongku Lee, Ahmad Ali
DOI
:10.4103/pm.pm_589_21
Background:
Glycation refers to the interaction between sugars and proteins leading to the production of harmful advanced glycation end-products (AGEs). Many different diseases such as neurodegenerative, cardiovascular, and secondary complications in diabetic patients have reported the involvement of these products. Diabetes is one such disease, wherein the accumulation of AGEs causes secondary complications.
Objectives:
This study inclines to investigate the antiglycation and antioxidant potential of aqueous and methanolic extracts of
Tribulus terrestris
(TT).
Materials and Methods:
The
in-vitro
glycation system (bovine serum albumin [BSA] and glucose) was incubated with aqueous and methanolic extracts of TT for 28 days at 37°C. Standard methods such as browning, nitroblue tetrazolium (NBT) assay, 2,4-dinitrophenylhydrazine (DNPH) assay, and assessment of fluorescent AGEs were carried out spectroscopically to check the number of glycation products. The antioxidant activity was also assessed for both extracts. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and agarose gel electrophoresis were also performed.
Results:
The presence of aqueous extracts (AE) of TT showed the inhibition of carbonyl content (41.74%) and total AGE generation (40.14%). Reduction in β-amyloid aggregation was observed in SDS-PAGE analysis and thioflavin-T assay. The conformation of DNA was maintained in the presence of aqueous extract in glycated DNA in the presence of Fe
+
.
Conclusion:
The extracts of fruits of TT exhibit potential antiglycation and antioxidant activity
in-vitro
in protein and DNA. It may be used as a therapeutic agent for the management of diabetes and its complications.
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Goniothalamin alleviates inflammation and arthritic markers in adjuvant-induced arthritis in rats
p. 348
Dong Yang, Guangfeng Niu, Jie Guo
DOI
:10.4103/pm.pm_257_21
Background:
Goniothalamin (GTN) is a well-known styryl lactone that possesses anti-inflammatory and anticancer properties. In this study, we evaluated the potential of GTN on experimental arthritis based on the complete Freund's adjuvant (CFA)-induced arthritis model in rats and the effect was compared with standard drug.
Materials and Methods:
The animals were divided into six groups: normal, CFA, CFA + indomethacin (10 mg/kg bw), CFA + GTN (50 mg/kg bw), CFA + GTN (100 mg/kg bw), and GTN (100 mg/kg bw) alone. The antiarthritic activity of GTN was measured by paw thickness, arthritic score, body weight, organ weight, biochemical markers, inflammatory mediators, and histopathologic analysis of ankle joints.
Results:
GTN significantly reduced the paw thickness, arthritic score, body weight changes, and organ weight. It reduced the levels of C-reactive protein, alkaline phosphatase, rheumatoid factor, and proinflammatory cytokines in a dose-dependent manner. It also minimized the histopathologic changes in ankle joints induced by CFA in a dose-dependent manner.
Conclusion:
These results demonstrate that GTN is effective against inflammation and is an alternative agent for the management of arthritis.
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Growth stage-specific Accumulation of Cardiac Glycosides in Two Variants of
Calotropis gigantea
(L.) W.T. Aiton
p. 353
Pankaj Singh, Yatendra Singh, Sanjeev Kanojiya, Vineeta Tripathi, Dipak Kumar Mishra
DOI
:10.4103/pm.pm_525_21
Background:
Calotropis gigantea
(L.) W.T. Aiton, belonging to the family Apocynaceae, is a source of many bioactive cardiac glycosides. Accumulation of secondary metabolites in plants depends upon various external and internal factors, including the plant age or growth stage.
Objectives:
The main aim of this study was to identify the correct variant and growth stage of
C. gigantea
for the maximum yield of targeted metabolites.
Materials and Methods:
In the present experiment, three different tissues, that is, leaf, stem, and root, of 3–12-month-old white flower variant (WFV) and purple flower variant (PFV) of
C. gigantea
were investigated to find out the growth stage-specific accumulation pattern of five cardiac glycosides (CGs) and their three genins.
Results:
Highest concentrations of calotropin, uscharin, and uscharidin were found in the stem of five-month-old WFV. Frugoside and uzarigenin were at peak levels in the stem of a nine-month-old member of the same variant. Calotropagenin and coroglaucigenin were at maximum levels in the root and leaf of 8- and 10-month-old members, respectively, of this variant too. The only CG accumulated at maximum level in the root of 11-month-old PFV was asclepin and its quantity was comparable to 8-month-old WFV root.
Conclusion:
It was observed that the WFV was superior to PFV, considering the accumulation of most of the CGs. Therefore, it can be concluded that the specific tissues of 5–10-month-old members of WFV are optimum for an economic yield of respective CGs.
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Evaluation of apoptotic and cytotoxic effect of robinin in TPC-1 and SW1736 human thyroid cancer cells
p. 360
Yawei Shen, Periyannan Velu, Xin Huang, Taoping Dang
DOI
:10.4103/pm.pm_270_21
Background:
During the past few years, thyroid cancer (TC) has increased in terms of rate of morbidity and mortality. Plant flavonoids have shown positive effect in regulating thyroid tumorigenesis via inhibition of apoptosis. Robinin is a natural flavone glycoside isolated from
Vinca erecta
with potent pharmacological activities.
Materials and Methods:
In this study, we aimed to explore the apoptotic and cytotoxic activity of robinin against TPC-1 and SW1736 cells.
Results:
Robinin (20 μM/mL) significantly suppressed the growth and cell proliferation and induced apoptotic activity in TC cells. According to the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, TPC-1 and SW1736 cancer cells revealed 100% cell viability. Robinin inhibited growth of TC cells in a dose-dependent manner. It was found that reactive oxygen species (ROS) generation in TPC-1 and SW1736 cancer cells was decreased; however, robinin (20 μM/mL) enhanced ROS formation. The study of apoptosis in TPC-1 and SW1736 cells revealed morphological changes and damaged nuclei. Robinin (20 μM/mL) triggered a powerful apoptosis signal and caused loss of membrane integrity in TC cells. It also increased the activity of caspases 8 and 9. Robinin (20 μM/mL) decreased the levels of Bcl-2, c-Myc, and cyclin-D1 and increased the levels of Bax and caspase-3 when compared to control and robinin-treated cells. It exhibited potent antiproliferative and apoptotic activity in TC cells.
Conclusion:
Robinin can be useful in the treatment of thyroid cancer.
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Molecular effect of betanin on the molecular expression pattern of cell proliferative and inflammatory signalling pathways in DMBA-induced oral carcinogenesis in hamsters
p. 366
Yingying Fan, Ge Ma, Qingyan Guo, Jian Wang, Periyannan Velu
DOI
:10.4103/pm.pm_281_21
Background:
Cancer development is a sequential process as a result of various cellular adaptation events. Oral cancer (squamous cell carcinoma) is the most predominant variety of head and neck cancer. Betanin (BTN) is isolated from beetroot extracts and is a highly bioavailable antioxidant. BTN exerts a chemopreventive and cytotoxic activity on numerous cancer cells. However, precise identification of the molecules responsible for this tumor-inhibitory effect is pending. This study aimed to understand the molecular mechanisms underlying the chemopreventive effects of BTN in 7,12-dimethylbenz (a) anthracene (DMBA)-induced oral cancer in experimental hamsters.
Materials and Methods:
The interactions of BTN with antioxidant enzymes, lipid peroxidation, apoptosis, and inflammatory markers in the presence of DMBA were investigated in male golden Syrian hamsters.
Results:
Oral supplementation of BTN treatment (50 mg/kg BW) daily to oral tumor-bearing rats successfully prevented DMBA-induced oral carcinogenesis. Furthermore, BTN administration significantly prevented weight loss and reduced the tumor occurrence, burden, volume, and biochemical parameters such as TBARS, LOOH, SOD, CAT, GPx, GSH, and vitamins E and C. The histological analysis and expression pattern of molecular markers (increased apoptosis (caspase-3 and 9), proliferative markers (PCNA and Cyclin-D1), and inflammatory markers (TNF-α and COX-2) investigated in hamsters' buccal mucosa tissues revealed a significant anti-tumorigenic nature of BTN.
Conclusion:
The findings of this study show that BTN markedly reduces DMBA-induced oral cancer in hamsters.
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β-Caryophyllene reduced oxidative stress and expression of apoptotic markers in streptozotocin-induced diabetic rats
p. 373
Binghong Hua, Yin Xiao, Fuling Li
DOI
:10.4103/pm.pm_331_21
Purpose:
In this study, we aimed to investigate the effects of β-caryophyllene against oxidative stress-induced apoptosis in the animal model of diabetes.
Materials and Methods:
Experimental diabetes was induced in the rat model through the administration of streptozotocin. Plasma samples were tested for the lipid profile. Plasma insulin levels were measured by performing electrochemical immunoassay. Fasting blood glucose was measured using the glucometer. Intracellular levels of reactive oxygen species (ROS) were measured through dichlorodihydrofluorescein diacetate method. Markers of oxidative stress such as catalase (CAT), reduced glutathione (GSX), malondialdehyde (MDA), and superoxide dismutase (SOD) levels were determined by using the colorimetric kits. Expression of apoptotic proteins such as Bax, Bcl-2, Pro-caspase 9, caspase 9, and β-actin was analyzed using the Western blot technique.
Results:
According to our results, β-caryophyllene normalized the body weight of diabetic rats and improved the lipid profile of the experimental animals. It also normalized the levels of fasting blood glucose. Moreover, the plasma insulin levels significantly increased after the administration of β-caryophyllene. The β-caryophyllene treatment caused a significant decline in the ROS and MDA levels together with an increase in the levels of CAT, SOD, and GSX levels in diabetic rats. Interestingly, the markers of apoptosis such as Bax and caspase 9 decreased after the administration of β-caryophyllene in the neural tissue of diabetic rats. However, the levels of Bcl-2 protein were increased in the β-caryophyllene-treated rats.
Conclusion:
β-Caryophyllene exhibits significant antidiabetic effect, which might be due to its capacity to reduce oxidative stress and inhibit apoptotic markers in the peripheral neural tissue of diabetic rat. These results point toward the potential of β-caryophyllene in the treatment of diabetes.
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Effect of
Lycium Barbarum
polysaccharide combined with cisplatin on the proliferation of human lung cancer cells
p. 378
Yunfan Liu, Yuemei Du, Liping Gao
DOI
:10.4103/pm.pm_354_21
Background:
Lycium barbarum
polysaccharide (LBP) is a water-soluble polysaccharide extracted from
Lycium barbarum
. LBP exhibits potential pharmacological activity, including anti-cancer activities. However, so far, the effect of LBP in combination with cisplatin (DDP) on the proliferation of human alveolar adenocarcinoma cell line (A549) of non-small cell lung cancer (NSCLC) has not been reported.
Objectives:
In this study, we aimed to investigate the effect of LBP combined with DDP on the proliferation of A549 cells.
Materials and Methods
: The cells were divided into four groups: control group, DDP group, LBP group, and LBP and DDP combined group and Three parallel experiments were set up in each group. The survival rate of A549 cells and the effects of DDP and LBP alone or in combination was detected using the cell counting kit-8 (CCK-8) method. The activity of superoxide dismutase (SOD) was determined by xanthine oxidase method. The content of glutathione (GSH) was determined by colorimetry. The content of malondialdehyde (MDA) was determined by thiobarbituric acid method. The apoptosis, cell cycle phase, and the level of reaction oxygen species (ROS) formed were detected by flow cytometry. The expression of apoptosis-related proteins such as Bcl-2, Bax, and caspase-3 and cell cycle–related proteins namely, CDK4, cyclin D1, and p-Rb were detected via Western blot analysis.
Results:
DDP (≥ 6 mg/L) and LBP (≥ 8 mg/L) alone significantly inhibited the proliferation of A549 cells (
P
< 0.01), and LBP combined with DDP significantly enhanced the inhibitory effect on the proliferation of A549 cells (
P
< 0.01). DDP significantly reduced the activity of SOD and the level of GSH (
P
< 0.01), and significantly increased the level of MDA and ROS (
P
< 0.01). Compared with DDP group, the activity of SOD and the content of GSH and MDA in LBP and DDP combined group did not change significantly (
P
> 0.05), but the content of ROS decreased significantly (
P
< 0.01). DDP and LBP, alone and in combination, significantly promoted the cellular apoptosis (
P
< 0.01). They also significantly downregulated the expression of Bcl-2 and upregulated the expression of Bax and caspase-3, and the level of Bax/Bcl-2 was significantly increased (
P
< 0.01). DDP blocked the A549 cells in S phase, whereas LBP blocked the cells in G2/M phase. DDP and LBP combination significantly downregulated the expression of cell cycle regulatory proteins namely, CDK4, cyclin D1, and p-Rb (
P
< 0.05).
Conclusion:
LBP in combination with DDP inhibited the proliferation of A549 cells, and this combined effect is related to the expression of apoptosis-related proteins and the regulation of cell cycle mediated by cyclin D1-CDK4-Rb pathway.
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Natural anti-hepatitis B virus flavones isolated from schimperi vatke growing in Saudi Arabia: Cell culture and molecular docking study
p. 386
Sarfaraz Ahmed, Mohammad K Parvez, Komal Zia, Mohammad Nur-e-Alam, Zaheer Ul-Haq, Mohammed S Al-Dosari, Adnan J Al-Rehaily
DOI
:10.4103/pm.pm_600_21
Background:
Stachys schimperi
Vatke has been previously reported for its analgesic, antipyretic, antioxidant, antimicrobial and cardioprotective properties.
Objectives:
Phytochemical analysis and assessment of anti-hepatitis B virus (anti-HBV) activity of
S. schimperi
.
Materials and Methods:
Surface extraction was performed to isolate the phytoconstituents using chromatographic techniques, including HPLC. The isolates were identified by a 1D and 2D NMR spectroscopic data. Further, the isolates were tested for cytotoxicity using an MTT assay. Non-cytotoxic doses of the isolates were assessed for their antiviral potential on cultured HepG2.2.15 cells. To rationalize the plausible mechanisms of the tested anti-HBV active compounds, molecular docking studies were carried out using HBV polymerase (Pol) enzyme.
Results:
The NMR data proved the structure of isolates as artemetin [5-hydroxy, 3' 3′,4′'6,7-penta methoxy flavone] (1), chrysosplenetin [5,4′-dihydroxy, 3'3′' 6,7-tetra methoxy flavone] (2) and calycopterin [5,4′-dihydroxy, 3' 6,7'8-tetra methoxy flavone] (3). Notably, this is the first report on the isolation of these three compounds from
S. schimperi
as well as artemetin and calycopterin from the genus
Stachys.
Further antiviral assessment of the non-cytotoxic dose showed marked inhibitions of HBV antigens (HBsAg/HBeAg) by artemetin (52.28%/46.52%) and calycopterin (61.24%/57.26%) in HepG2.2.15 cells. Chrysosplenetin, however, did not show any anti-HBV activity. Artemetin and calycopterin exhibited anti-HBV activity, possibly through inhibition of HBV-Pol as revealed by molecular docking.
Conclusion:
We report the identification of
anti-HBV active
flavones artemetin and Calycopterin from
S. shimperi
. Our data strongly warrant further molecular and pharmacological studies on artemetin and calycopterin toward developing potential anti-HBV therapeutics.
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Ruanmailing oral liquid inhibits atherosclerosis in ApoE
−/−
mice via regulation of TGF-β1/SMAD4 signaling pathway
p. 393
Tianmin Wu, Xuanbin Huang, Wenjuan Xue, Liufang Fan, Wenyan Xie, Youzhen Huang, Jinshui Chen
DOI
:10.4103/pm.pm_410_21
Background:
In this study, we aimed to investigate the effect of Ruanmailing oral liquid on atherosclerosis and transforming growth factor (TGF)-β1/SMAD4 signaling pathway in apolipoprotein E-knockout (ApoE
−/−
) mice induced by a high-fat diet.
Materials and Methods:
A total of 40 ApoE
−/−
mice were randomly divided into five groups: control group, model group, low-dose group, high-dose group, and Lipitor group. Mice fed with standard diet formed the control group. ApoE
−/−
mice exhibited high-fat diet-induced atherosclerotic phenotype. The other four groups were high-fat diet model groups, low- and high-dose Ruanmailing groups (1.75 and 4.55 mL/kg/day, respectively), and Lipitor group (3.0 mg/kg/day). After 12 weeks of administration, the levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein-cholesterol (LDL-C), and high-density lipoprotein-cholesterol (HDL-C) were measured by blood sampling from the orbital vein of the mice, and the pathological changes in thoracic aorta due to atherosclerosis were observed by hematoxylin and eosin (H and E) staining. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the concentration of serum TGF-β1, and reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analysis were performed to detect the expression of
SMAD4
and
GATA2
in the thoracic aorta of mice in each group.
Results:
Compared with the high-fat model group, the level of serum lipids in the test group were reduced (
P
< 0.01 or
P
< 0.05) and the ratio of plaque area to luminal area (W/L) was significantly reduced (
P
< 0.05). The pathological examination indicated that the atherosclerotic lesions in the thoracic aorta of ApoE
−/−
mice were alleviated, and the high-dose Ruanmailing group had the most significant anti-atherosclerotic effect.
Conclusion:
Ruanmailing oral liquid exhibited an anti-atherosclerotic effect, and its mechanism may be related to the intervention of GATA2 in the TGF-β1/SMAD4 signaling pathway to reduce the differentiation and proliferation of arterial smooth muscle cells.
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Celastrus paniculatus
seed oil ameliorates oxidative stress in lipopolysaccharide-induced respiratory inflammation in mice
p. 400
William R Surin, Eveline J Sharon, Archana Kullu, Santhosh Kacham, S Sandya
DOI
:10.4103/pm.pm_575_21
Background:
Celastrus paniculatus
have been reported to possess various medicinal properties. However, very little is known about its effect on respiratory inflammation.
Objectives:
We evaluated
Celastrus paniculatus
seed oil (CP oil) on lipopolysaccharide (LPS)-induced mice model of respiratory inflammation.
Materials and Methods:
Both short-term and long-term studies were carried out. In short-term study, C57BL/6 mice were exposed to LPS (10 mg/kg) by intranasal route for one week. CP oil (1 g/kg) was given orally on the 1
st
, 3
rd
and 5
th
day. All the mice were sacrificed on the 7
th
day. In long-term study, C57BL/6 mice were exposed to LPS (10 mg/kg) by the intranasal route for one month. CP oil (1 g/kg) or theophylline (1 mg/kg) was administered twice a week for one month. At the end of the treatment, Bronchoalveolar lavage (BAL) fluid was collected and labelled with CD3e FITC, Ly-6G (Gr-1) PE-Cy-7, Allergin-1 eFlour® 660, CD14 APC and 2',7'-dichlorofluorescein diacetate (20 μM) and analyzed by flow cytometer (BD FACSCanto™ II) for cellular infiltration and reactive oxygen species (ROS) generation. There was a significant reduction in neutrophil infiltration following CP oil treatment for one month ( 68±6% vs 40±7%,
P
< 0.05). Furthermore, there was a significant decrease in ROS generation following CP oil treatment for one week (5873 ± 1133 vs 2581 ± 1359;
P
< 0.05) and one month (20618 ± 1854 vs 5850 ± 1006;
P
< 0.05). Further, theophylline treatment for one month reduced the ROS generation significantly (20618 ± 1854 vs 5286 ± 2413;
P
< 0.05).
Conclusion:
Celastrus paniculatus
seed oil seems to reduce oxidative stress by inhibiting the generation of reactive oxygen species in LPS-induced lung inflammation.
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Exploring the potential quality marker of traditional tibetan medicine
Erigeron multiradiatus
based on ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry analysis, network pharmacology, and experimental verification
p. 406
Amu Hamamozhi, Jing Li, Xuyan Su, Wenfu Cai, Riza Zhao, Zhifeng Zhang
DOI
:10.4103/pm.pm_344_21
Background:
Erigeron multiradiatus
(EM), a well-known traditional Tibetan medicine, has been used for hundreds of years to treat various chronic metabolic diseases. However, previous studies on EM have primarily focused on its pharmacognosy, identification, and clinical efficacy, which lacks a systematic material basis research to identify therapeutic targets.
Objectives:
In this study, we investigated the potential quality marker (Q-marker) and the pharmacological mechanisms of EM during treatment of diabetic microangiopathy (DM) using both ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS/MS) and network pharmacology.
Materials and Methods:
The chemical profile of EM was qualitatively identified for the first time by using UHPLC-QTOF-MS/MS. A network pharmacology methodology was then used to predict the major active components and key pharmacological pathways of EM in the treatment of DM. The compound-target network was constructed using Cytoscape software. Finally, virtual screening was conducted using molecular docking, and a cytopathological model was established by culturing vascular endothelial cells under
in vitro
conditions to verify the predicted bioactive markers.
Results:
A total of 26 compounds were identified and tentatively characterized in the chemical profile of EM based on the reference standards and mass spectral data. Network analysis was performed on 107 overlapping gene symbols, and seven bioactive constituents (e.g., quercetin, apigenin, luteolin, scutellarin, rutin, chlorogenic acid, and chrysoeriol) were identified as potentially bioactive markers. The results of molecular docking showed that quercetin, scutellarin, and rutin exhibited higher binding affinity to core targets than that of the remaining four compounds. The effects of quercetin, scutellarin, and rutin on vascular endothelial cells were investigated under
in vitro
conditions. According to the results, scutellarin and quercetin significantly inhibited the proliferation of vascular endothelial cells and decreased the levels of nitrous oxide, and reduced glutathione.
Conclusion:
This study showed that scutellarin and quercetin can be the Q-markers of EM due to their possible therapeutic efficacy in treating DM. It provided fundamental insight into chemical profiling, pharmacological mechanisms, and quality control of EM.
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Structural characterization and emulsifying properties of a water-soluble
Buchanania lanzan
gum polysaccharide
p. 418
Saptarshi Samajdar, K Jayaram Kumar
DOI
:10.4103/pm.pm_96_22
Background:
Buchanania lanzan
gum, better known as chironji gum, has been traditionally used as a binder in food preparations. Water-soluble polysaccharides have good emulsifying properties, but the polysaccharide structure elucidation of water soluble chironji gum has not been carried out, and also, its use in emulsion has not been explored.
Objectives:
The aim of this study is to perform structural characterization of water-soluble polysaccharide isolated from chironji gum (CGPS) and explore its nanoemulsifying properties.
Materials and Methods:
The water-soluble polysaccharide CGPS was isolated and fractionated by Sepharose 6B column. Structural characterization was investigated by high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and gas chromatography–mass spectrometry (GC–MS). CGPS-stabilized nanoemulsion was formulated using Miglyol 812, Cremophor RH40, and diclofenac sodium and was evaluated.
Results:
The polysaccharide fraction had a yield of 33.85% ± 0.88% w/w. HPLC analysis revealed that CGPS consisted of galactose, rhamnose, arabinose, and glucose. CGPS had an average molecular weight of 201 kDa. Analytical studies, including NMR and GC–MS, showed that CGPS had repeating units of → 4)-α-d-Gal
p
(1 → 2)-α-l-Rha
p
-(1 → with terminal β-d-Glc
p
(1 → and α-l-Ara
f
(1 → residues. The nanoemulsions prepared with CGPS showed a droplet size ranging from 31.41 ± 0.26 to − 65.30 ± 0.21 nm, and negative zeta potential values were obtained for all the nanoemulsions.
In vitro
drug release studies of the CGPS nanoemulsion formulation revealed that diclofenac sodium release was delayed in simulated colonic fluid.
Conclusion:
This study indicates immense potential of CGPS for colon-targeted drug delivery system.
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Combination of platelet bio-specific extraction and high-performance liquid chromatography with diode-array detection/liquid chromatography-mass spectrometry method for analyzing platelet-targeted compounds in rhizoma chuanxiong
p. 427
Qian Zhang, Yuxiu Yang, Yao Xiao, Fenqing Wang, Yuanjia Hu, Zhining Xia, Fengqing Yang
DOI
:10.4103/pm.pm_390_16
Context:
Platelet is an important pharmacological target as it participates in complex processes of coagulation and hemostasis. Excessive platelet aggregation is responsible for the formation of pathogenic thrombi in patients with atherothrombotic disease.
Objective:
To expedite the search for platelet-targeted active candidates in Rhizoma Chuanxiong (root of
Ligusticum chuanxiong
Hort [
Umbelliferae
]).
Materials and Methods:
Rhizoma Chuanxiong ethyl acetate extract (EAE) was obtained by refluxing extraction method. A platelet bio-specific extraction combined with high-performance liquid chromatography with diode-array detection/liquid chromatography-mass spectrometry (HPLC-DAD/LC-MS) approach was employed to screen and identify the platelet-targeted compounds in Chuanxiong EAE, and the results were confirmed by
in vitro
antiplatelet aggregation test using turbidimetry method. Finally, HPLC-DAD analysis was employed for the quantitative determination of these hit compounds in Chuanxiong EAE.
Results:
A total of five hit components were tentatively detected and identified by HPLC-DAD/LC-MS, two of which, senkyunolide A and (Z)-ligustilide, were confirmed their antiplatelet activity by
in vitro
platelet aggregation experiment (both with IC50 <3 μg/mL, 2.42 μg/mL [12.60 μM] and 2.97 μg/mL [15.63 μM], respectively). Meanwhile, the quantitative results indicated that those two strongest inhibitors are the main components in Chuanxiong EAE (87.891 ± 1.668 mg/g and 109.058 ± 1.383 mg/g, respectively).
Conclusion:
Senkyunolide A and (Z)-ligustilide are the main active antiplatelet compounds in Rhizoma Chuanxiong EAE. The proposed platelet bio-specific extraction combined with HPLC-DAD/LC-MS method is of value for the discovering potential bioactive components in natural products.
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Protective effect of fermented
Cudrania tricuspidata
fruit extracts on acute rat reflux esophagitis
p. 435
Li Nan, Hyeon-Hwa Nam, Byung-Kil Choo
DOI
:10.4103/pm.pm_481_21
Background:
Reflux esophagitis (RE) is a type of gastroesophageal reflux disease (GERD) that severely reduces the quality of life. The
Cudrania tricuspidata (C. tricuspidata
) fruit is commonly used in food and has medicinal values such as clearing heat, relaxing muscles, and collaterals.
Objectives:
We evaluated fermented
C. tricuspidata
fruit extract (FCt) in this study. We measured its anti-inflammatory activity by using LPS-induced Raw 264.7 cell model and evaluated its role in improving acute reflux esophagitis in rats.
Materials and Methods:
The gastrodin content before and after fermentation was determined. NO production in cell supernatant was measured using the Griess reagent method, and TNF-α and IL-1β secretion was detected using the Quantikine ELISA kit. Expression level of proteins was measured by western blotting. Immunofluorescence assay was used for confirming the activation of NF-κB. The protective effects of FCt on RE were conducted in the rat RE model by observing esophageal tissue damage and inflammatory proteins expression levels in the esophagus.
Results:
The results showed that the content of gastrodin in
C. tricuspidata
fruit increased upon fermentation. FCt inhibited the production of NO, TNF-α, and IL-1β and the activation of NF-κB. In addition, FCt significantly improved esophageal tissue damage caused by acid reflux and inhibited the expression of inflammatory proteins iNOS and COX-2 in esophageal tissue. The term of tight junction (claudin-4) is increased to maintain the barrier function of cell tissues.
Conclusion:
We suggested that FCt could consider being a candidate medicine material for treating RE.
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Combined network pharmacology and cell experiments to explore the anti-inflammatory mechanisms of matrine
p. 443
Lichao Wu, Fangfang Tao, Tengfei Sun, Junhui Zhao, Junfeng Li, Wenhong Liu
DOI
:10.4103/pm.pm_431_21
Background:
Matrine is an alkaloid compound isolated from the root of
Sophorae Flavescentis Radix
. Despite being a potent anti-inflammatory agent, the underlying mechanisms of these anti-inflammatory properties have not been scrutinized.
Objectives:
To briefly elucidate the anti-inflammatory mechanism of matrine through network pharmacology and
in vitro
experiments.
Materials and Methods:
Multiple public databases analyzing targets associated with matrine and inflammation were surveyed. Thirty common targets acquired from the Venn diagram were used to compose the protein–protein interaction (PPI) network visualized by Cytoscape software for the subsequent analysis.
Results:
Network pharmacology revealed that 30 genes in matrine could interrupt the inflammatory metabolism to alleviate its development and progression. The cellular experiments revealed that matrine reduced the expression of interleukin-6 and tumornecrosis factor-α.
Conclusion:
Our results provide preliminary evidence and theoretical background regarding the role of matrine to assuage the inflammatory reaction, and support its future clinical application.
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Improvement effect of evodiae fructus water extract against DSS-induced ulcerative colitis
p. 450
Min Ju Kim, Jin A Lee, Mi-Rae Shin, Ji Hye Lee, Hae-Jin Park, Sanghyun Lee, Seong-Soo Roh
DOI
:10.4103/pm.pm_465_21
Background:
Evodiae fructus is a natural plant that is used as a medicinal material in many Asian countries such as Korea and China. However, little is known about its effect on ulcerative colitis.
Objectives:
Thus, the purpose of this study was to find the improvement effect of Evodiae fructus in mice model with dextran sulfate sodium (DSS)-induced colitis.
Materials and Methods:
The animal experiments were conducted using ulcerative colitis animals induced by DSS. To induce ulcerative colitis in BALB/c mice used in the experiment, 5% DSS was supplied as drinking water for 7 days. BALB/c mice were divided into five groups (
n
= 8): Group 1 (normal group), Group 2 (5% DSS control group), Group 3 (5% DSS + sulfasalazine 60 mg/kg treated), Group 4 (5% DSS + Evodiae fructus water extract 50 mg/kg treated), Group 5 (5% DSS + Evodiae fructus water extract 100 mg/kg treated). After the animal experiment, the expression of antioxidant, oxidative stress, inflammatory, and necrosis-related factors in colonic tissues was analyzed by western blotting.
Results:
As a result, the Evodiae fructus treatment group reduced oxidative stress by increasing the expression of Nrf2, an antioxidant-related transcription factor. Also, the expression of NF-κB, an inflammatory transcription factor, was reduced, thereby reducing the expression of pro-inflammatory proteins and inflammatory cytokines.
Conclusion:
These results suggest that Evodiae fructus treatment has an inhibitory effect on inflammation against DSS-induced colitis. Therefore, Evodiae fructus has the potential to be used as a natural medicine for ulcerative colitis.
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Quercetin of
Potentilla bifurca
3-Glycosylation Substitution Impact the Inhibitory Activity on α-glucosidase
p. 458
Xuanzhao Qu, Jinping Li, Peiying Yan, Guiyao Wang, Wei Liu, Yang Zeng, Likuan Liu
DOI
:10.4103/pm.pm_522_21
Background:
Inhibition of glycosidase activity is an effective diabetes therapy and prevention strategy. In this study, we aimed to explore the effects of quercetin and its derivatives extracted from
Potentilla bifurca
on α-glycosidaseinhibited vitality.
Materials and Methods:
Various chromatographic methods and separation techniques were used to separate quercetin and quercetin derivatives. An enzyme reaction system was established to determine the inhibitory activity of
P. bifurca
compounds against α-glucosidase and the type of enzyme inhibitor reaction.
Results:
The changes in activity between quercetin and its derivatives is observed in the following areas: (1) The action of flavonoids is expected to be influenced by the substituents of different molecules. Glycosylation of quercetin reduces its hypoglycemic effect, and the molecule created after adding penta-glycoside is larger than that formed after adding hexa-glycoside. (2) The acylated drugs can possibly reduce their activity.
Conclusion:
P. bifurca
has been shown to inhibit glycosidase activity, making it effective in diabetes treatment.
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Hecogenin attenuates isoproterenol-induced myocardial infarction through nuclear factor-kappa B-mediated signaling pathway in rats
p. 463
Ruihui Pei, Chaolong Duan, Annamalai Vijayalakshmi
DOI
:10.4103/pm.pm_264_21
Background:
Cardiac-related problems accounts for about 10%–35% of all deaths. Medicinal plants and their derivatives are a rich source of effective cardioprotective agents. Hecogenin (HCG) is a sapogenin with a wide spectrum pharmacological property. In this study, we aimed to evaluate the cardioprotective effect of HCG against isoproterenol (ISO)-induced myocardial infarction (MI) in rats.
Materials and Methods:
The animals were divided into four groups. Group I: Control group consisted ofrats fed onstandard pellet (23 days); Group II: HCG group, HCG 50 μg/kg body weight (BW), oral administration, (P. O) on 21 days; Group III: ISO group, 60 mg ISO/kg BW; subcutaneous (sc) administered on days 22 and 23; Group IV: HCG (50 μg/kg BW. P. O) (21 days) + ISO (days 22 and 23). After sacrificing the animals, we analyzed thebiochemical and molecular markers.
Results:
Our results showed that there were no significant differences in the BW of rats; however, the HCG and ISO groups showed a significant reduction in heart weight. When compared with ISO group, hepatic and cardiac biomarkers were low in HCG + ISO group. Furthermore, compared to the ISO group, the level of lipid peroxidation products was restored to their optimal level in the HCG + ISO group. According to the histopathological findings, animals from HCG group demonstrated restoration of their tissue architecture. Immunohistochemistry demonstrated that animals in the HCG group showed reduced expression levels of nuclear factor kappa B (NF-κB) and p53, which was similar to the control and HCG group. This demonstrates cardioprotective effects of HCG.
Conclusion:
HCG attenuated ISO-induced MI via inhibition of activation of NF-κB and p53 signaling pathway. Overall, the findings recommended that HCG is a promising therapeutic agent for the treatment of MI.
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Neuroprotective activity of
Datura inoxia
and
Turnera diffusa
extracts in an
in vitro
model of neurotoxicity
p. 470
Jesús Vélez-Huerta, Mónica A Ramírez-Cabrera, Omar González-Santiago, Juan M de J. Favela-Hernández, Eder U Arredondo-Espinoza, Isaías Balderas-Rentería
DOI
:10.4103/pm.pm_543_21
Background:
Due to their therapeutic properties and easy access, in Mexican culture, plants are used to treat several diseases. This is the case of
Datura inoxia
and
Turnera diffusa
, which affect their central nervous system with psychotropic, anticholinergic, analgesic, and stimulating properties.
Objectives:
In this research, extracts D1 (methanolic), D2 (chloroformic), and D3 (hexanoic) from
D. inoxia
and T1 (ethanolic), T2 (chloroformic), and T3 (hexanoic) from
T. diffusa
were evaluated to determine their neurotoxic and neuroprotective activity
in vitro
.
Materials and Methods:
Neurotoxic activity was evaluated in PC-12 cells exposed to the extracts at concentrations ranging from 3.12 to 200 μg/mL. The neuroprotective activity was evaluated by pretreating PC-12 cells with the extracts and subsequently exposing them to neurotoxic compound glutamate. In both assays, cell viability was evaluated with 4-(3-[4-iodophenyl]-2-[4-nitrophenyl]-2H-5-tetrazolio)-1,3-benzene disulfonate (WST-1). Antioxidant activity was studied with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, and antiapoptotic activity was evaluated with the caspase-3 fluorescent assay.
Results:
T. diffusa
was more neurotoxic than
D. inoxia
, and both plants showed neuroprotective activity at low concentrations.
T. diffusa
had more antioxidant activity than
D. inoxia
. Neither plant had antiapoptotic activity.
Conclusion:
Extract D1 of
D. inoxia
and T1 of
T. diffusa
showed neuroprotective activity, and both extracts had the lowest neurotoxic effect
in vitro
.
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Molecular cloning and quantitative real-time PCR analysis to study the expression of tryptophan decarboxylase gene from chillies (
Capsicum annuum
L.) against whitefly
p. 476
Niranjanadevi Jeevanandham, Murugan Marimuthu, Senthil Natesan, Karthikeyan Gandhi, Nalini Prabakar, Pasupathi Elango, Mohan Chinnarasu, Visvanathan Maarthandan
DOI
:10.4103/pm.pm_362_21
Background:
One of the amazing and economical spices is Chilli (
Capsicum annum
L.) and also called “wonder spice,” and it gives great significance to traditional household use and export as well as such a remarkable cash crop in India. However, the important constraints in chilli production are chilli leaf curl disease which has been caused by whitefly (
Bemisia tabaci
) transmitted by begomoviruses and leads to a major decline in yield.
Aim:
To explore the stability of the insect resistance gene (tryptophan decarboxylase [TDC]) in resistant genotypes after being challenged with a known number of whiteflies. Since, biotic stress defense allied mechanism regulated during tryptophan biosynthesis.
Materials and Methods:
The presence of endogenous genes was verified using the polymerase chain reaction (PCR), cloning (T/A vector), and gene expression level (quantitative real-time PCR).
Results:
Amplification of mitochondrial
coxI
gene fragment using the primer (C1-J-2195 and L2-N-3014) produced
B. tabaci
specific ~ 800 bp band. Besides, the whitefly sequence was aligned with the NCBI blast where 92% of identity was observed. TDC gene expression was greater in P2 accession leaves at 48 h post-infestation with 20 number of
B. tabaci
after feeding and downregulation in less TDC gene expression level at moderately resistant accession ACC 20. Further, all the amplified gene sequences were aligned using NCBI BLAST. The expression of TDC genes in chillies could demoralize the
B. tabaci
fitness causes mortality.
Conclusion:
The satisfactory control of viral disease may be achieved with the application of molecular biology, DNA, and RNA-based technologies include the cloning of insect resistance genes.
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Luteolin suppressed growth of colon tumor via inflammation, oxidative stress, and NLRP3/IL-1β signal axis
p. 494
Qi Yao, Yuan Luo, Lingjia Sun, Hongxia Wang, Wenjie Li
DOI
:10.4103/pm.pm_284_21
Background:
Luteolin, a natural flavonoid, exerts anticancer activities. In this study, we evaluated the role of luteolin in colon cancer, and its underlying mechanisms.
Materials and Methods:
The effect of luteolin on the proliferation of human colon tumor (HCT)-116, SW480, and Caco-2 cells was evaluated. Subsequently, the effect of luteolin on the proliferation, migration, and invasion of HCT-116 cells was monitored. Then, we analyzed the level of inflammation and oxidative stress in the mouse model transplanted with HCT-116 under
in situ
conditions. Furthermore, we investigated whether luteolin attenuates growth of colon tumor under
in vivo
conditions.
Results:
Our results demonstrate that luteolin reduced the proliferation of HCT-116, SW480, and Caco-2 cells. In addition, it blocked the migration and invasion of HCT-116 cells. Moreover, luteolin increased the production of interleukin (IL)-2/10 and decreased the production of IL-6, interferon-β, tumor necrosis factor-α, and IL-1 β. Furthermore, it increased the level of superoxidase dismutase and glutathione peroxidase and decreased the levels of malondialdehyde significantly. Luteolin significantly lowered the sensitization of NOD-leucine-rich repeat and pyrin-containing protein 3 (NLRP3)/caspase-1/IL-1 β signal axis. Luteolin exerted the above actions in a dose-dependent manner.
Conclusion:
The results of this study indicate that luteolin mitigates the growth of colon cancer via suppressing the inflammatory processes, oxidative stress, and NLRP3/IL-1 β signal axis. It might serve as a promising candidate for colon cancer.
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Excision and incision wound healing activity of apigenin (4',5,7-trihydroxyflavone) containing extracts of
Carissa carandas
Linn. fruits
p. 502
Chanchal Kumar Mishra, Ashok Kumar Pattnaik, Pankaj Dagur, Dhiraj Kumar, Manik Ghosh
DOI
:10.4103/pm.pm_100_21
Background:
Carissa carandas
Linn. (family Apocynaceae) is a climbing shrub, with a height of 10 or 15 ft (3–5 m). It is found in Jharkhand and other states of India. It is commonly known as “Karunda” or “Jasmin flower Carissa.” The major requirement for its growth is full exposure to the sun.
C. carandas
blooms and fruits throughout the year.
Objectives:
Gel of fruit extracts was formulated to determine its wound healing ability by applying it topically on wounds in Wistar rats.
Materials and Methods:
To study the wound healing activities, excision and incision studies, histopathology, and phytochemical studies were done. Ethanolic fruit extract of
C. carandas
was applied topically as 20% w/w gel. Excision and incision wound healing models were employed for determining wound contraction and tensile strength percentage.
Results:
The ethanolic fruit extracts of
C. carandas
(EFCC) 10% and 20% w/w showed effective activity compared to the simple ointment treated and untreated groups. EFCC 20% w/w had a better effect compared to EFCC 10% w/w. There was an increase in the rate of contraction of wounds on 18
th
post-wounding day (3.04 ± 0.55***) and increase in tensile strength (388.31 ± 0.59***). Tissue pathology of revitalized skin of EFCC revealed increased production of collagen, mononuclear cells, and fibroblasts. Apigenin (4',5,7-trihydroxyflavone) was also isolated from the extract.
Conclusion:
The EFCC ointment (20% w/w) showed a significant increase in wound healing activity, validating its use by the tribal people.
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Effects of total flavonoids extracted from
Chromolaena odorata
Linn. on immunosuppression: A network pharmacology-based and experimental study
p. 510
Yao Hu, Fan Yang, Zhen Lin, Aftab Qamar, Di-Hua Zhu, Meng-Meng Chen, Min Zhang, Qing-Lang Song, Dan-Ju Kang, Hong-Ying Lin, Zhi-Bao Chen, Jin-Jun Chen
DOI
:10.4103/pm.pm_545_21
Background:
Chromolaena odorata
Linn. (CO) is a perennial herb that is enriched with flavonoids and exhibits immune regulatory activities. For these characteristics, CO can be used as a potential immunoregulator based on the immunosuppression state.
Objectives:
The aim of this study was to assess the effects of flavonoids extracted from CO (FCO) on immune regulation and evaluate their mechanism of action by network pharmacology (NP), followed by
in vivo
confirmation.
Materials and Methods:
FCO were ultrasonically extracted through immersion in alcohol. The potential targets were predicted using a “FCO–immunosuppression–target” network. When the functional enrichment analyses were conducted, a mice model was employed to demonstrate the effects. The hematological indexes and serum levels of immunoglobulin G (IgG), immunoglobulin M (IgM), interferon-γ (INF-γ), interleukin 2 (IL-2), and tumor necrosis factor-α (TNF-α) were measured. The variations observed in immune organs and the changes in expressions of INF-γ, T-box transcription factor 21 (Tbx-21), and GATA Binding Protein 3 (GATA3) were reported.
Results:
NP results showed that a total of 198 targets of FCO were involved in immunosuppression. As indicated by the functional analysis results, FCO impacted T helper (Th) cell differentiation, which might be a vital functional pathway underlying its immune regulatory effects. During animal experiments, the values of hematological indexes, serum levels of IgG, IgM, and TNF-α, and the immune organ indexes increased in FCO groups, and the relative mRNA expressions of INF-γ and Tbx-21 and less damage of the spleen and thymus were reported.
Conclusion:
FCO impacts Th1 and Th2 differentiation pathways and assists in immunosuppression by regulating the secretion of various cytokines and the expression of associated genes, which demonstrates FCO as a promising natural immunoregulator.
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