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Jan-Mar 2021
Volume 17 | Issue 73
Page Nos. 1-208

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ORIGINAL ARTICLES  

Physalis peruviana juice and seeds methanolic extracts; gas chromatography mass spectrometry; antioxidant and anticancer against human A549, HepG2 Highly accessed article p. 1

DOI:10.4103/pm.pm_262_20  
Objectives: Physalis peruviana L. is a medicinal herb and its consumption increases annually in The Middle East, also the scientific research on it increases due to its valuable nutrient. Materials and Methods: Methanolic extracts of P. peruviana L. seeds and juice were screened for their anticancer and antioxidant. Gas chromatography-mass spectroscopy profiling was performed for all extracts. . Results: The identification of seeds and juice methanolic extract showed the main sex compounds; ethanol, 2,2'-oxybis-, caffeic acid in both of the extracts. Octadecadienoate ethyl and octadecenoic acid have been found in seed extract, and octadecadienoic acid and ferulic acid were in juice extract. Seeds extract has phenolic and flavonoid content as 53.58 and 45.56, respectively, comparing to juice extract (26.58 and 7.30, respectively). The antioxidant activities of seeds extract using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant potential (FRAP) (28.73 at 50 μg/ml and 1164.10, respectively) comparing to juice extract values (4.06 at μg/ml and 848.43, respectively). Conclusion: The conspicuous optimistic result is that seeds extract showed cancer inhibition against human HepG2 and A549 (81.45 and 85.34, respectively) comparing to juice extract (44.06 and 32.06, respectively). Therefore, the demand to increase the usage of Physalis or golden berry in people's diet is a demand to face the environmental oxidative stress.
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Biological and chemical evaluation of some African plants belonging to Kalanchoe species: Antitrypanosomal, cytotoxic, antitopoisomerase I activities and chemical profiling using ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometer p. 6

DOI:10.4103/pm.pm_232_20  
Background: Human African trypanosomiasis is one of the most serious neglected tropical diseases causing fatal symptoms and death. Natural products are a main source for anti-infective metabolites. Objectives: The objective of the study is to evaluate eight different plants belonging to the Kalanchoe species growing in Egypt for antitrypanosomal, antimalarial, antileishmanial, cytotoxic, and antimicrobial activities. Materials and Methods: The antitrypanosomal activity against Trypanosoma brucei; cytotoxic activities against human colon carcinoma, human hepatocyte carcinoma, and human breast adenocarcinoma cell lines; antileishmanial activity against Leishmania donovani; antimalarial activity against Plasmodium falciparum; and antimicrobial activities of all plant extracts have been examined. As well as the identification of the secondary metabolites for the most active extract was performed via ultra performance liquid chromatography coupled to high resolution quadrupole time of flight mass spectrometer operated in negative and positive ionization modes. Results: Among the examined plant extracts, Kalanchoe longiflora leaves extract showed promising activity against T. brucei with an inhibition concentration of sample at 50% fall in absorbance (IC50) value of 17.6 μg/mL. K. longiflora with other extracts exhibited promising cytotoxic activities. Profiling of the polar secondary metabolites of K. longiflora revealed the presence of 47 metabolites including 31 flavonoids, 9 phenolic acids, 4 anthocyanidins, 2 chalcone glucoside, and 1 coumarin. To determine the mechanism of action of K. longiflora extract as a potent antitrypanosomal and cytotoxic agent, we investigate its ability to inhibit topoisomerase I enzyme. K. longiflora extract showed an excellent activity with an IC50 value of 0.148 μg/mL. Conclusion: These interesting results open the door for further research aiming at the development of a successful treatment for Trypanosoma from K. longiflora.
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Spectrum–effect relationships between fingerprints of radix Polygoni multiflori-Achyranthes bidentate and antiosteoporosis effect based on different extraction solvents p. 16

DOI:10.4103/pm.pm_258_20  
Background: Polygonum multiflorum and Achyranthes bidentate are traditional Chinese herbs, which show significant antiosteoporosis (OP) effect. These two herbs are commonly used as a combination in traditional Chinese medicine. Materials and Methods: In this study, the left humerus section of the mice were stained with hematoxylin and eosin staining to establish the spectrum-effect relationships between the high-performance liquid chromatography fingerprint of different solvent extracts and the anti-OP effect. Finally, we analyzed the correlation between the content of main compounds of different solvent extracts and the anti-OP effect. Results: The compatibility had obvious therapeutic effect on OP induced by retinoic acid. The results of pharmacodynamic analysis showed that the extract had a certain effect on the bone-specific alkaline phosphatase, the level of tartrate-resistant acid phosphatase-5b (TRACP-5b), and estrogen 2 and had a great influence on TRACP-5b especially. The results of spectrum-effect relationships and correlation analysis showed that the level of β-ecdysterone and tetrahydroxystilbene is negatively correlated with bone resorption index and TRACP-5b. The higher the content of the two compounds in the combination, the better the effect of treating mouse OP induced by retinoic acid. Conclusion: It is suggested that the compatibility of P. multiflorum and A. bidentata can be used to treat OP, and their mechanism of action is through reducing osteoclastic effect. β-ecdysterone and tetrahydroxystilbene may be the active components of combination of Polygoni multiflori-A. bidentate.
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Vasorelaxant and antioxidant activity of some medicinal plants from Campeche, Mexico p. 23

DOI:10.4103/pm.pm_291_20  
Context: Brosimum alicastrum, Cnidoscolus chayamansa, Tradescantia spathacea, Turnera diffusa, Manilkara zapota, and Jatropha gaumeri are medicinal plants recognized in Mexican Mayan Culture. Aim: Methanol leaves extracts of these plants were use as raw material to develop a phytochemical, spectroscopy, and pharmacological analysis. Subjects and Methods: Methanol maceration was carried out and were compared in terms of yield extraction, chlorophyll, simple phenolic and flavonoids content, antioxidant activity (DPPH and β-Carotene bleaching models), as well as isolated aorta rings (E+), precontracted with noradrenaline. Results: Best content of simple phenolic and flavonoids compounds was recorder in B. alicastrum, J. gaumeri and T. diffusa. J. gaumeri extract exert an antioxidant (β-carotene bleaching: EC50: 0.8 ± 0.1 μg/mL, Emax: 85.7% ± 0.4%; DPPH: EC50: 60.3 ± 1.8 μg/mL, Emax: 60.4% ± 1.8%; P < 0.05) and vasorelaxant (EC50: 161.61 ± 7.45 μg/mL; Emax: 79.71% ± 3.88%; P < 0.05) activity in a concentration dependent-manner. Fourier transform infrared spectroscopy analysis allowed estimating a 1.26 and 2.28% of quercetin (Q) and gallic acid (GA) in J. gaumeri. GA exerts antioxidant activity in DPPH model (EC50: 1.6 ± 0.2 μg/mL; Emax: 92.9% ± 3.3%) and Q/GA (1:2) mixture improves inhibition of β-carotene bleaching (EC50: 0.005 ± 0.005 μg/mL; Emax: 69.2% ± 0.7%; P < 0.05). Conclusion: J. gaumeri is a medicinal plant employed in Mayan traditional medicine and GA and Q could be related to traditional uses, as well as responsible for the pharmacological effects. GA and Q interactions improve inhibition β-Carotene bleaching activity, which suggests greater solubility in lipophilic systems and potential interactions at the plasma membrane level.
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The inhibitory effects of different kinds of ginsenosides on skin pigmentation in melasma mice model p. 31

DOI:10.4103/pm.pm_9_20  
Objectives: The study was aimed to compare the effects of seven different kinds of ginsenosides and total saponins on melasma and to explore their related mechanisms. Materials and Methods: The inhibitory rate of ginsenosides on tyrosinase (TYR) activity was measured in vitro. The skin cream comprising different kinds of ginsenosides was prepared and the melasma mice model was established. After successful modeling, different skin creams were applied to the backside of mice for 30 days and these mice were separated into 11 groups. Afterward, the melanocytes and melanin in the epidermal cells were analyzed under the microscope. Finally, the activity of superoxide dismutase (SOD), TYR and the content of malondialdehyde (MDA) in melasma mice were determined by different assay kits. Results: The outcomes of in vitro experiment showed that ginsenosides could affect TYR activity with different concentrations. In the melasma mice model, ginsenosides and total saponins inhibited the growth of melanocytes and the synthesis of melanin. Furthermore, with the increasing use of skin cream comprising ginsenosides and total saponins, the activity of SOD showed an increase trend, while MDA content and TYR activity exhibited a decrease trend. Thirty (30) days of treatment with ginsenosides (except for Rg1 and Rg2) and total saponins significantly increased the SOD activity and decreased the MDA and TYR levels compared with those in melasma mice without treatment (P < 0.05). Conclusion: Skin cream comprising ginsenosides Rb1, Rc, Rd, and Re could employ better protective roles in melasma by suppressing oxidative stress and inhibiting the synthesis of melanin.
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Caffeic acid treatment augments the cell proliferation, differentiation, and calcium mineralization in the human osteoblast-like MG-63 cells p. 38

DOI:10.4103/pm.pm_186_20  
Background: Osteoporosis is an imperative health problem that extremely distresses the public that leads to a higher risk to the bones from both spontaneous and accidental bone fractures. Caffeic acid is a polyphenol compound that happens naturally in numerous vegetables such as coffee beans, potatoes, propolis, olives, and carrots with many pharmacological aids. Objectives: The current study was planned to examine the potential of Caffeic acid in proliferation, differentiation, and calcium mineralization of osteoblast-like MG-63 cells. Materials and Methods: The cell viability of Caffeic acid-supplemented MG-63 cells was examined through the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity test. The mRNA expression of alkaline phosphatase (ALP) and osteocalcin was reviewed through reverse transcription polymerase chain reaction study. The calcium deposition rate in the Caffeic acid-treated MG-63 cells was studied through Alizarin Red S (ARS) staining. Results: The result of the MTT test exposed that the 10 and 20 mg/kg of Caffeic acid supplementation did not show any cytotoxicity to the osteoblast-like MG-63 cells; instead, it helped the viability of MG-63 cells. The expression of ALP is particularly increased in the Caffeic acid-supplemented MG-63 cells, whereas the osteocalcin expression was noticeably diminished. The ARS staining was exhibited that the Caffeic acid treatment was noticeably enhanced the calcium mineralization rate in the osteoblast-like MG-63 cells. Conclusion: Based on the findings of investigation, it was proved that the Caffeic acid treatment was significantly enhanced the cell proliferation, differentiation, and calcium mineralization in the osteoblast-like MG-63 cells. Hence, it was clear that Caffeic acid can be engaged as the potential agent for the purpose of bone regeneration.
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Dieckol attenuates cell proliferation in Molt-4 leukemia cells via modulation of JAK/STAT3 signaling pathway p. 45

DOI:10.4103/pm.pm_2_20  
Background: Leukemia is a cancer of the hematopoietic stem cells, which leads to an uncontrolled proliferation of leukocytes in blood. It is responsible for one of the most important cancer-associated deaths across the globe. Materials and Methods: In this study, we analyzed whether dieckol (DEK), a polyphenolic compound obtained from brown algae, can suppress cell proliferation via regulation of JAK/STAT3 signaling pathway in leukemia cell lines (Molt-4). Results: According to our results, DEK induced cytotoxicity, altered the cell morphology, caused nuclear damage, enhanced the formation of reactive oxygen species, decreased the production of mitochondrial membrane potential, reduced the levels of antioxidants (reduced glutathione, catalase, and superoxide dismutase), and augmented the level of thiobarbituric acid reactive substances in Molt-4 cell lines. Furthermore, STAT3 has been recognized as an important transcriptional mediator that controls cell proliferation. Thus, suppression of STAT3 transcription is a novel approach for the suppression of Molt-4 cell proliferation. In this study, DEK inhibited STAT3 translocation, thereby suppressing the increased expression of cyclin E1, PCNA, cyclin D1, and JAK1 in Molt-4 cell lines. Conclusion: In summary, DEK suppressed the cell proliferation of Molt-4 cells via inhibition of JAK/STAT3 signaling pathway.
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Immunomodulatory effect of d-carvone in swiss albino mice with benzo(a)pyrene-induced lung cancer p. 51

DOI:10.4103/pm.pm_508_19  
Background: Globally, lung cancer is the second most cause of deaths, which accounts for approximately 29% of the cancer cases worldwide. d-Carvone is considered a vital constituent of many essential oils with immense pharmacological benefits. Objective: In this study, we examined the immunomodulatory effect of d-carvone in the Swiss albino mice model against benzo(a)pyrene (BaP)-induced lung carcinoma. Materials and Methods: BaP was orally administered to the mice (50 mg/kg body weight [bw] for 4 weeks [twice a week]). Post-BaP administration, d-carvone (20 mg/kg bw) was orally administered to the mice (20 mg/kg bw). We calculated the tumor incidence and performed the following measurements: lung and bw, hematological parameters, immune complexes (phagocytic and avidity indexes, nitroblue tetrazolium reduction, soluble immune complex levels, immunocompetent cells (leukocytes, lymphocytes, and neutrophils), immunoglobin (Ig) levels (IgG, IgA, and IgM), level of xenobiotics and enzymes that point toward hepatic dysfunction, carcinoembryonic antigen (CEA), and proinflammatory cytokines (PICs) in experimental and normal mice. The level of oxidative stress in the experimental animals was investigated. The lung tissues of investigational animals were examined for the histopathological changes. Results: According to the results, there was an increased level of lipid peroxidation and decreased level of antioxidant activity in the lung tumor samples. The counts of lymphocytes, polymorphonuclear cells, and macrophages were notably decreased and increased, respectively, by the d-carvone post-treatment. Moreover, BaP-induced inflammation that is indicated by the increased in the level of PICs and CEA proteins in lung samples. d-Carvone attenuated the levels of PICs and CEA. Conclusion: The results of this study reveal that d-carvone prevented the cells against BaP-induced inflammation in lung cancer without causing adverse effects.
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β-caryophyllene, a natural bicyclic sesquiterpene, induces apoptosis by inhibiting inflammation-associated proliferation in MOLT-4 leukemia cells p. 58

DOI:10.4103/pm.pm_550_19  
Background: Leukemia is known to be a common type of cancer mostly affecting children. The standard therapeutic treatment available for leukemia has many drawbacks with serious side effects. Therefore, plant-based chemotherapeutic agents that show less/no toxic side effects might be an efficient way to treat leukemia. Therefore, in this study, we aimed to explore the potential of β-caryophyllene, obtained from various plants sources, and found that it persuades oxidative stress-associated apoptosis during the repression of inflammation and proliferation in MOLT-4 leukemia cancer cells. Materials and Methods: In this study, MOLT-4 cells were incubated with β-caryophyllene (15 and 20 μM) for 24 h and found that β-caryophyllene increased the level of cytotoxicity and reactive oxygen species (ROS) and decreased the level of antioxidants, mitochondrial membrane potential, and apoptotic reaction in MOLT-4 cells. Cell proliferation and apoptosis are important cellular events, and inhibition of cell proliferation along with the generation of proapoptotic marker has been considered as a novel task for treatment of cancer. Results: According to our results, β-caryophyllene induced apoptosis by downregulating the expression of Bcl-2 family of proteins and upregulating the expression of caspases involved in BAX-associated apoptosis in MOLT-4 cells. It also downregulated the expression of biomarkers involved in proliferation (proliferating cell nuclear antigen and cyclin-D1) and inflammation (tumor necrosis factor-α, interleukin-6, nuclear factor-kappa B, and cyclooxygenase-2). Conclusion: In summary, β-caryophyllene potentially induced apoptosis by generating ROS and by inhibiting inflammation and proliferative genes in MOLT-4 leukemia cells.
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Chemical composition and neurotherapeutic potential of Glaucium corniculatum extracts p. 67

DOI:10.4103/pm.pm_348_20  
Background: The discovery of natural agents that suppress neural apoptosis is of strategic importance for the treatment of neurodegenerative diseases. Glaucium corniculatum has been used as a traditional medicine because of its memory-enhancing and neuroprotective properties. However, there has not been a study about this plant's apoptosis suppressive properties until now. Objectives: The present study examines the possible suppressive effects on apoptosis of three different G. corniculatum extracts (chloroform, methanol, and water) in neuronal-differentiated PC12 (dPC12) cells induced by hydrogen peroxide (H2O2) and presents the first association of compound content differences in extracts with the suppressive property. Materials and Methods: The chemical composition of these extracts was analyzed by Fourier transform infrared spectroscopy (FTIR) and gas chromatography-mass spectroscopy. The changes caused by the extracts in apoptotic markers were examined by real-time quantitative polymerase chain reaction and Western blot. Results: Extracts contained alkaloid, fatty acid, alcohol, alkane hydrocarbon, ester, ether, alkane, alkene, aliphatic, aromatic, and phenolic compounds with different relative proportions. In all the three extracts, the major compound was allocriptopine. Oleamide and 2-monopalmitin were the major fatty acids, respectively, identified in water and methanol extracts. Extracts were suppressed H2O2-induced apoptosis via decreasing the expression of Bax, Caspase-3 and -9, while increasing Bcl-2, in a concentration-dependent manner. The water extract showed the highest apoptosis suppression while methanol was the second. Conclusion: G. corniculatum extracts suppress apoptosis in dPC12 cells and allocriptopine, oleamide, and 2-monopalmitin may be responsible of apoptosis suppressive effect.
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Rehmannia glutinosa polysaccharide increases the expression of erythropoietin and vascular endothelial growth factor in rats with chronic renal failure by activating hypoxia-inducible factor-2α p. 76

DOI:10.4103/pm.pm_13_20  
Background: To investigate the effect of Rehmannia glutinosa polysaccharide (RGP) in rats with chronic renal failure. Materials and Methods: In this study, we established a rat model of adenine-induced chronic renal failure. Hemoglobin (Hb), red blood cells (RBCs), total protein (TP), serum albumin (ALB), blood urea nitrogen (BUN), cystatin C (Cys C), and serum creatinine anhydride (SCA) were detected in the blood samples of rats. Hematoxylin and eosin staining was used to observe the pathological changes. Immunohistochemical was used to detect the protein expression of hypoxia-inducible factor (HIF)-2α, erythropoietin (EPO), and vascular endothelial growth factor (VEGF) and calculate the value of microvessel density (MVD) in renal tissues. M1001 and GPRRP + PT2385 groups were added to further validate the mechanism of RGP. Results: In the case of rats with chronic renal failure, GPRRP and EPO contributed to the repairment of renal tissue. The level of Hb, TP, and ALB, as well as RBC counts was significantly increased, and the renal function indexes, including BUN, SCA, and Cys C, were significantly decreased, and HIF-2α, EPO, VEGF, and MVD were significantly increased. The effects observed in M1001 group was similar to those in GPRRP group, but compared with GPRRP + PT2385 group, the biochemical indicators and renal function indexes were significantly decreased, which means HIF-2α inhibitor antagonized the protective effect of RGP. Conclusion: RGP played an essential role in repairing the renal injury in rats with chronic renal failure. The therapeutic mechanism of action may be related to the activation of HIF-2α, thereby increasing the expression of EPO and VEGF and then reducing renal anemia and renal tissue damage.
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An edible bioactive fraction from Rosa multiflora regulates adipogenesis in 3T3-L1 adipocytes and high-fat diet-induced C57Bl/6 mice models of obesity p. 84

DOI:10.4103/pm.pm_175_20  
Objectives: Rose varieties are cultivated worldwide for ornamental flowers and have been greatly valued in the cosmetics and medicinal applications. Medicinal preparations from the different parts of the rose, including leaves, petals, and fruits, have been studied for various health benefits. Here, we have studied the efficacy of a standardized rose petal extract using the experimental models. Materials and Methods: The anti-obesity effect of a polyphenol-rich extract from the petals of Rosa multiflora var. platyphylla (RoseFit, 2%–3% isoquercetin) was studied using in vitro adipocyte differentiation model and in vivo diet-induced obesity model. Results: Treatment with 250 and 500 μg of rose fit dose-dependently reduced the differentiation of 3T3-L1 adipocytes and lipid accumulation thereof. The expression of adipogenic markers such as CCAAT/enhancer-binding protein-alpha and peroxisome proliferator-activated receptor gamma was markedly down regulated in the adipocytes treated with RoseFit. A 6-week treatment of high-fat diet (HFD) fed mice with RoseFit (100 mg and 200 mg/kg) significantly reduced the body weight, liver and fat pad weights, and adipocyte cell size of epididymal fat. Rose fit treatment also mitigated the hepatic inflammation significantly as compared to untreated HFD mice. Conclusion: Collectively, our findings suggest that the standardized rose petal extract may be used as a functional ingredient in food to increase the nutritional value and health-promoting effects.
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Targeted metabolomic analysis of seven short-chain fatty acids in feces of rats with spleen-deficiency syndrome after administering raw and bran-fried atractylodis rhizoma by gas chromatography-mass spectrometer p. 93

DOI:10.4103/pm.pm_185_20  
Background: For treating spleen-deficiency syndrome (SDS), atractylodis rhizoma (AR) is usually treated with bran, which is measured to enhance efficacy. Newly, we found that bran-fried AR (BFA) had benefits over raw AR in refining gut flora. However, the change about metabolism of gut flora caused by raw and BFA remains unclear. Aim: The aim of this study was to compare the difference between raw and BFA acting on short-chain fatty acid (SCFA) metabolism in SDS rats, a metabolic pathway comprising gut flora. Materials and Methods: The model of SDS rats was recognized and then given raw and BFA by gavage. The serum was collected for the determination of motilin (MTL), gastrin (GAS), trypsin (TRY), and amylase (AMS) by enzyme-linked immunosorbent assay. The feces were collected for the determination of 7 SCFAs under selective ion monitoring mode by gas chromatography-mass spectrometer. Results: Raw and BFA can upsurge the levels of MTL, GAS, TRY, and AMS in the serum of SDS rats, diminution the levels of acetic acid and propionic acid, and rise the level of hexanoic acid in the feces of SDS rats, and BFA showed noteworthy activity than raw AR. Conclusion: Raw and BFA can improve the acetic acid, propionic acid, and hexanoic acid metabolism in SDS rats, and BFA was more potent than raw AR.
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Antiproliferative and apoptosis inducing effect of delphinidin against human bladder cancer cell line p. 101

DOI:10.4103/pm.pm_548_19  
Background: Urinary bladder cancer is one of the leading causes of death worldwide. This study was conducted to evaluate the effect of delphinidin on proliferation, apoptosis, and cell cycle arrest in the urinary bladder cancer cell line (T24). Materials and Methods: We performed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay to analyze the cytotoxic effect of delphinidin on T24 cell proliferation. We tested different concentrations of delphinidin. Furthermore, the effect of delphinidin on cell cycle arrest was studied using flow cytometric assays and the percentage of apoptotic cell death was estimated via annexin V/propidium iodide. Results: According to our results, delphinidin (10–60 μg/mL) demonstrated a significant cytotoxic effect against T24 cells. Morphological analysis and analysis of the oxidative stress revealed a significant number of dead cells in comparison to the control group. The results of flow cytometry revealed that delphinidin caused dose-dependent apoptosis of T24 cells. Conclusion: In summary, delphinidin can be a promising option for the treatment of urinary bladder cancer in the future.
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Identification of differentially expressed genes and biological pathways in sanguinarine-treated ovarian cancer by integrated bioinformatics analysis p. 106

DOI:10.4103/pm.pm_111_20  
Aim: This study was intended to identify potentially target genes and underlying biological pathway of sanguinarine in ovarian cancer. Methods: We obtained the expression changes of downstream target genes and underlying biological pathways regulated by control and sanguinarine groups via Affymetrix gene expression profile chip in ovarian cancer cells. An Affymetrix Genechip Agilent mRNA Array was used to recognize differentially expressed genes (DEGs). Afterward, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed for the DEGs using the DAVID database. Results: A total of 1185 DEGs between sanguinarine and control groups were identified, including 835 upregulated and 350 downregulated DEGs. The result of GO analysis recommended that the DEGs were mostly enriched in biological processes, including negative regulation of gene expression, nitrogen compound metabolic process, and transcription from RNA polymerase II promoter. Alterations in cellular components (CC) were suggestively enriched in the cytoskeleton and endoplasmic reticulum. The changes in molecular function were suggestively enriched in nucleic acid-binding transcription factor activity, protein dimerization activity, and enzyme binding. The results of the KEGG pathway analysis indicated that the DEGs were mostly concentrated in “Systemic lupus erythematosus,” “MAPK signalling pathway,” “Pathways in cancer,” and “Metabolic pathways.” Conclusion: The present study provided insights into the mechanism underlying sanguinarine target genes in ovarian cancer cells, which might be used as effective targets for OC diagnosis and treatment.
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Therapeutic effect of Huzhangoside D in rats with knee osteoarthritis induced by anterior cruciate ligament transection p. 112

DOI:10.4103/pm.pm_298_20  
Background: Knee osteoarthritis (KOA) is an age-related disease. Huzhangoside D is a saponin isolated from genus Clematis L. (Ranunculaceae). The aim of this study is to explore the anti-inflammatory, apoptotic, and autophagy regulation effects of huzhangoside D on KOA in a rat model. Materials and Methods: The KOA model was established by an anterior cruciate ligament transection surgery. Huzhangoside D was administered for 4 weeks. The weight-bearing assay, morphology observation, and intrinsic mechanism exploration were performed. Results: After administration, the weight-bearing assay showed that huzhangoside D promoted joint function recovery. Hematoxylin-eosin and safranin O-Fast green staining indicated that huzhangoside D ameliorated the structural damage. The Mankin scores were decreased in the huzhangoside D groups. Huzhangoside D enhanced cartilage thickness. Enzyme-linked immunosorbent assay study revealed that huzhangoside D downregulated the proinflammatory cytokine (tumor necrosis factor alpha, interleukin-6, and interleukin-1β) levels, while it upregulated the anti-inflammatory cytokine (interleukin-10) level in rat serum. Terminal deoxynucleotidyl transferase dUTP nick end labeling assay showed that huzhangoside D downregulated the apoptosis ratio of cartilage cells. Immunohistochemical staining showed that huzhangoside D upregulated the autophagy-related protein beclin-1, ATG5, ATG7, and light chain 3 levels and downregulated the p62 level. Moreover, the AKT and mTOR signaling pathway activities were downregulated. The 3-MA combination with huzhangoside D downregulated the weight-bearing function and morphology of the knee and upregulated the proinflammatory cytokines, which showed the role of autophagy as a protective mechanism in the effect of huzhangoside D. Conclusion: This study revealed that huzhangoside D is a promising agent in KOA treatment.
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Identification and characterization of compounds from methanolic extracts of Launaea procumbens by gas chromatography-MS, liquid chromatography-electrospray ionization-MS/MS, and ultra-performance liquid chromatography-electrospray ionization-quad time of flight/MS p. 120

DOI:10.4103/pm.pm_333_20  
Background: Launaea procumbens is an important medicinal plant took its place also in the ingredient of food as preparation of sherbet, leaves in making curries, and as a goat fodder in most of the part of India. Objectives: The present work was aimed at investigating the phytochemicals of methanolic extract of L. procumbens leaves and its isolated mixtures from column fractions. Materials and Methods: The method was successfully developed and two new compounds (chlorogenic acid and 7-hydroxyflavanone) by liquid chromatography (LC)-electrospray ionization (ESI)-MS/MS and four new compounds (lactucin, isorhametin, 1-monopalmitin, and 1-hexacosanol) by ultra-performance liquid chromatography (UPLC)-ESI-quad time of flight (QTOF)/MS in L. procumbens were identified. Identification of compounds by LC-ESI-MS/MS and UPLC-ESI-QTOF/MS was identified based on the accurate mass of pseudomolecular [M+H]+ ion tandem mass spectrometry (MS/MS) data and by comparing retention times, mass spectra, and molecular weights with those published in the literature. Results: The six new compounds, namely chlorogenic acid, 7-hydroxyflavanone, lactucin, isorhametin, 1-monopalmitin, and 1-hexacosanol were identified by in L. procumbens. Conclusion: A total of 26 compounds by gas chromatography-MS and five compounds by LC mass-ESI tandem mass spectrometry (LC-ESI-MS/MS) and seven compounds by UPLC-ESI tandem mass spectrometry/quadrupole-time-of-flight-mass spectrometry (UPLC-ESI-QTOF/MS) technique were identified.
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Anticancer activity of abietane diterpenoids from Salvia libanoticum grown in Lebanon p. 127

DOI:10.4103/pm.pm_265_20  
Background: The Salvia plant and their metabolites are well reported for their valuable therapeutic effects and as potential remedies for treatment of many diseases. Salvia libanoticum is an endemic species to Lebanon where its metabolites have never been investigated. Objectives: The objectives were to evaluate the potential of abietane diterpenes from the roots of Salvia libanoticum as anticancer agents and explore some essential chemical features. Materials and Methods: Crude extract from the roots of Salvia libanoticum was separated using chromatographic techniques and spectroscopic analysis. The anticancer activities of the isolated compounds along with the crude extract were evaluated against MDA-MB-231 breast cancer cells and HCT116 human colon cancer cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: Eight abietane diterpenes were isolated to be royleanone (1), 6,7-dehydroroyleanone (2) orthosiphonol (3), horminone (4), 7α-acetylhorminone (5), taxoquinone (6), 8,9-epoxy-7-oxoroyleanone (7), and 7-oxoroyleanone (8). All compounds including the extract revealed dose-dependent inhibitory effects that varied between the two cell lines, indicating cell-type specificity and suggesting different cell-compound interactions. Discussion: The most effective compound was found to be 7α-acetylhorminone, with IC50 of 18 and 44 μM on HCT116 and MDA-MB-132 cells, respectively. The results suggested that oxygenated C7 is essential for the cytotoxic activity. Moreover, the carbonyl group at position C7 leads to higher activity than the hydroxyl group. Conclusion: This study reported the potential of abietane diterpinoids as anticancer agents. It also suggested Salvia libanoticum and its diterpinoids as promising remedies in colon and/or breast cancer therapy. Further studies are needed to explore the exact interaction of these compounds with cancer cells at molecular level.
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Skin-healing properties of ginsenoside Rd against Ultraviolet-B-induced photooxidative stress through up-regulation of antioxidant components in HaCaT keratinocytes p. 134

DOI:10.4103/pm.pm_319_20  
Background: Korean ginseng (Panax ginseng Meyer) is a traditional herbal medicine used worldwide today. Ginsenoside Rd (Rd), one of its main ginsenosides, has been ascertained to have various pharmacological efficacies including neuroprotective and anti-inflammatory activities. Objectives: This work assesses the antioxidant and protective potentials of Rd against ultraviolet (UV)-B-induced skin photooxidative stress in HaCaT keratinocytes. Materials and Methods: Cell viability was detected using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Intracellular reactive oxygen species (ROS) were measured using dichlorodihydrofluorescein diacetate. Promatrix metalloproteinase-2 (proMMP-2) activity and protein were detected using gelatin zymography and western analysis. Total glutathione (GSH) content and total superoxide dismutase (SOD) activity were spectrophotometrically determined. Results: Rd, at varying concentrations non-toxic to HaCaT keratinocytes, attenuated the UV-B-induced ROS generation. Rd at 5, 12, and 30 μM attenuated the UV-B-induced proMMP-2 gelatinolytic activities to 59.3% ± 9.5%, 41.3% ± 9.5%, and 13.1% ± 8.9%, respectively, of those of the non-treated control cells. It could also diminish the UV-B-induced proMMP-2 protein levels. Rd at 5, 12, and 30 μM augmented the UV-B-reduced total SOD activities to 1.6 ± 0.2-, 2.4 ± 0.3-, and 3.2 ± 0.2-fold of those of the non-treated control cells, respectively. Rd could up-regulate the UV-B-reduced total GSH. Conclusion: Rd has counteracting properties against elevated ROS and proMMP-2 and attenuated GSH and SOD under UV-B irradiation, implying that it possesses a protective activity against photoaging possibly through up-regulating antioxidant components. These findings suggest that Rd can be considered as a novel natural resource for anti-photoaging functional cosmetics.
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Protective effects of ginsenoside Rh1 on intervertebral disc degeneration through inhibition of nuclear factor kappa-B signaling pathway p. 140

DOI:10.4103/pm.pm_579_19  
Background: In this study, we aim to explore the protective effect of ginsenoside Rh1 against intervertebral disc degeneration (IDD) and the related mechanism. Materials and Methods: IDD model in Sprague-Dawley rats was established and the animals were treated with different concentrations of ginsenoside Rh1 for 4 weeks, after this, the animals were sacrificed and the intervertebral disc of was collected for analysis using quantitative polymerase chain reaction. Western blot analysis was performed for quantifying the expression levels of glycosaminoglycans (GAGs) and Types I and Type II collagen. Moreover, serum samples were collected and the expression levels of some of the inflammatory cytokines such as interleukin (IL)-1 β and IL-6 were evaluated. Next, we collected the nucleus pulposus (NP) cells from the animals and were divided into five groups: control, IDD, treatment groups with different concentrations of ginsenoside Rh1 (10, 20, and 50 μg/mL). After treatment, the levels of IL-1 β and IL-6 in the cell culture supernatant were examined. Then, we performed western blot analysis to quantify the levels of B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-extra-large (BCL-xL), and nuclear factor kappa-B (NF-κB) in different groups. Results: We observed that ginsenoside Rh1 significantly downregulated the expression of type I collagen and upregulated the expression of type II collagen and GAG under in vivo conditions. Moreover, the expression levels of IL-1 β and IL-6 in the serum samples of IDD rats and cell culture supernatant of NP cells isolated from the IDD rats were significantly increased. However, ginsenoside Rh1 significantly increased the levels of Bcl-2 and Bcl-xL and decreased the levels NF-κB both under in vitro and in vivo conditions. Conclusion: Ginsenoside Rh1 demonstrated protective effect against the IDD via regulation of IL-1 β/NF-κB signaling pathway.
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Chemical profile and anxiolytic- and anticonvulsant-like effects of Miconia albicans (Sw.) Triana (Melastomataceae) leaves in adult zebrafish Highly accessed article p. 146

DOI:10.4103/pm.pm_176_20  
Background: Miconia albicans is a vegetable widely used in folk medicine as an alternative for the treatment of pain and inflammation. This study aimed to evaluate the chemical profile and anxiolytic- and anticonvulsant-like effects of the aqueous extract M. albicans leaves (CAVEL) in adult zebrafish. Materials and Methods: The antioxidant activity, chemical prospecting, and toxicity tests were performed. Then, the anxiolytic- and anticonvulsant-like effects were investigated in adult zebrafish. Results: It was detected the presence of flavonoids, xanthones, besides alkaloids, flavonoids, and polyphenols in CAVEL. Eighteen polyphenols and nine flavonoids were identified in CAVEL. CAVEL showed antioxidant activity and no cytotoxic potential. The effects of CAVEL in adult zebrafish were dependent on the route of administration: CAVEL (Per os [p.o.] and intraperitoneal [i.p.]) did not alter the locomotion of the animals, presented anxiolytic-like effect (p.o. and i.p.) and delayed anticonvulsant-like effect (p.o.). The anxiolytic-like effect was prevented by GABAergic receptor antagonist. M. albicans has pharmacological potential for the treatment of anxiety and anticonvulsant and these results support studies of isolation and characterization of bioactive principles. Conclusion: M. albicans has pharmacological potential for the treatment of anxiety and anticonvulsant and these results support studies of isolation and characterization of bioactive principles.
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Novel mathematical model for the assessment of similarity of chromatographic fingerprints of volatile oil from Houttuynia cordata p. 154

DOI:10.4103/pm.pm_192_20  
Background: The analysis of similarities among fingerprints of Chinese herbal medicines is an important quality control tool to determine the authenticity of the herbal medicines. Objectives: In this study, we aimed to develop a novel mathematical model to analyze the similarity of the chromatographic fingerprints of Houttuynia cordata (HC). Materials and Methods: Total quantum statistical moment similarity (TQSMS) expressions were deduced to evaluate the similarities between two chromatographic fingerprints. The volatile oil samples of HC were analyzed with gas chromatography-mass spectrometry, and the fingerprints were constructed by the area under the peak of the chromatograms. Results: There were nine peaks in common, and a total of 733 chemical constituents observed among 15 batches of samples. The number of peaks in the chromatographic fingerprints of the 15 batches of HC was 49–137, with a relative standard deviation (RSD) of 30.13%. The sum of area under the peak was 1.159 × 107–3.437 × 108 μv × s, with an RSD 174.56%; MCRTT was 9.410–18.602 min, with an RSD of 20.79%; and VCRTT was 37.549–81.504, with an RSD of 23.27%. The volatile oil composition and content of HC showed strong fluctuation. Therefore, its quality control from the variety and content of the components is impractical. Since TQSMS method can characterize the sample similarity, we can quantitate the correct probability of positive and negative conclusions regardless of the population origin of the samples. Conclusion: Our results show that TQSMS can be an additional method that can be used to assess the similarity of two chromatographic fingerprints.
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Bilobalide, a bioactive compound on sepsis-induced acute lung injury through its anti-inflammatory and antioxidative activity p. 163

DOI:10.4103/pm.pm_448_20  
Background: Sepsis, one of the major life-threatening conditions and responsible for approximately 40% of clinical Acute Lung Injury (ALI) patients. Nevertheless, there are no specific medications available to reduce mortality. Bilobalide, a natural bioactive component present in Ginkgo biloba, has several medicinal properties. However, the effect of Bilobalide against ALI residues unknown. And so, the therapeutic property and underlying molecular mechanism of Bilobalide was investigated against sepsis-prompted ALI in the murine model. Materials and Methods: In C57BL/6J mice model, sepsis introduced by cecal ligation and puncture (CLP) to induce lung injury. Results: Our results showed that Bilobalide treatment increased the survival rate of CLP-induced sepsis mice. Pretreatment with Bilobalide substantially inhibits the sepsis-induced lung exudation, which is exposed by measuring the wet/dry weight of lung and lung permeability of the mice. Also, Bilobalide attenuated the histopathological alterations alike alveolar hemorrhage and infiltration of inflammatory cells are assessed by hematoxylin and eosin staining, tumor necrosis factor-α, interleukin (IL)-1 β, IL-6 and macrophage inflammatory protein-2 pro-inflammatory mediators and myeloperoxidase are measured by enzyme-linked immunosorbent assay. Molecular mechanism of Bilobalide in lung inflammation whereby cyclooxygenase-2 (COX-2), induced nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), and the activation of the nuclear factor-kappa B (kB) (p65)/IkB are determined by immunoblotting technique. Moreover, pretreatment of Bilobalide significantly downregulated the expression of COX-2, iNOS, and phosphorylation of p65 and induced the IkB activation in the lung. Further, Bilobalide prevented the oxidative stress by upregulating the expression of HO-1 in lung tissues, and gene expression of sepsis-induced anti-oxidative enzymes (catalase, MnSOD, CuZnSOD, and GPx-1) in the Bilobalide-treated mice were induced dose-dependently and it determined by a quantitative-real time-polymerase chain reaction. Conclusion: Hence, we suggested Bilobalide has the ability to act as a possible therapeutic candidate against sepsis caused ALI.
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Garden cress (Lepidium sativum L.) seeds enhancing osteogenesis postinduced-bone fracture p. 170

DOI:10.4103/pm.pm_75_20  
Objectives: This study is aimed to evaluate the role of garden cress seeds (GCS) in osteogenic enhancement in bone fractures induced in rabbits. Materials and Methods: Thirty New Zealand White rabbits (Oryctolagus cuniculus) (n = 30) of 6 months of age and weighing 3–4 kg were included in this study. Rabbits were distributed into two main groups, One served as control and the other were subjected to induced transverse diaphyseal fractures of the left femurs. All rabbits were accommodated in cages and permitted to move freely without external support. Wound care, hygienic conditions, diet and behavior were observed and followed up on daily basis. At the end of the study, five rabbits of each subgroup were sacrificed, followed by dissection of the left femurs. Histomorphometric measurements were performed in all microscopic fields at a ×100 using by Leica microscope DM 2500 connected to a camera (Leica DFC 295) and Leica Q win V3 image analysis software. Results: Bone markers analysis revealed that the serum levels of osteopontin and Vitamin D in fractured femur rabbits fed on 6 g GCS showed a significant increase compared to those of untreated fractured femur by the end of the 2nd phase of the study. The serum levels of Osteocalcin in fractured femur rabbits fed on 12 g GCS showed a significant decrease compared to those of untreated fractured femur at the end of the study. The serum levels of Parathormone and Lactoferrin in fractured femur rabbits fed on 12 g GCS showed a significant increase compared to those of untreated fractured femur at the end of the 2nd and 3rd phases of the study accompanied by a significant elevation in liver function test serum levels of fractured femur rabbits fed on 6 g GCS at the end of the 2nd and 3rd phases of the study. The histomorphometric evaluation showed marked improvement of fractured femur rabbits fed on 6 and 12 g of GCS as compared to those of untreated fractured femurs. Conclusion: Garden cress seeds could be a promising alternative treatment in bone fracture.
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Salidroside induces apoptosis and autophagy in gastric cancer cells via regulation of mitogen-activated protein kinases signaling pathway p. 179

DOI:10.4103/pm.pm_119_20  
Background: Salidroside, an active ingredient of Rhodiola rosea, exhibits antiproliferative effect in gastric cancer (GC) cells. However, the involvement of salidroside in apoptosis and autophagy of GC cells has not been elucidated. Materials and Methods: Cell viability of BGC-823 cells was assessed by CCK-8 assay, and apoptosis was analyzed with terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling assay and flow cytometry. The apoptosis- and autophagy-associated proteins and mitogen-activated protein kinases (MAPKs) were determined by Western blot analysis. Autophagy was evaluated by green fluorescent protein-fused LC3 punctate formation. Results: Salidroside inhibited proliferation and promoted apoptosis and autophagy of BGC-823 cells in a dose-dependent manner. In addition, salidroside enhanced the phosphorylation of p38 MAPK, ERK1/2, and JNK. Furthermore, inhibition of MAPKs significantly abolished the effects of salidroside in BGC-823 cells. Conclusion: Salidroside induced apoptosis and autophagy of BGC-823 cells via activation of MAPK signaling pathway.
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Neuroprotective effect of neferine, an alkaloid against the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced Parkinson's disease mouse model p. 186

DOI:10.4103/pm.pm_291_19  
Background: Parkinson's disease (PD) is the second most chronic neurodegenerative disorders affecting the age-old peoples worldwide. Therefore, in the present study, we assessed the neuroprotective effect of neferine against the progressive Parkinson's induced mouse model. Mice were induced PD by treating them with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a neurotoxin selectively disrupts the dopaminergic neurons and then treated with neferine, a plant alkaloid. Materials and Methods: The neuroprotectant effect of neferine against MPTP-induced Parkinson's was assessed with behavioral analysis such as grid test and stride length measurement test, and the motor coordination was confirmed with rota rod test. Further, to assess the anti-inflammatory property of neferine, the levels of pro-inflammatory cytokines tumor necrosis factor-α, interleukin (IL)-1β and IL-6 were estimated. Results: To confirm the neuroprotectant effect of neferine, the levels of dopamine and the protein expression of inducible nitric oxide synthase, cylcooxygenase-2, rate-limiting enzyme of dopamine synthesis tyrosine hydroxylase in the substantia nigra tissue of control and experimental mice were evaluated. Our overall results authentically prove that neferine acts an anti-inflammatory agent decreased the levels of pro-inflammatory cytokines and increased the dopamine levels in substantia nigra, thereby protecting the mice from MPTP-induced Parkinsonism. Conclusion: The behavioral analysis also confirmed neferine as a persuasive neuroprotectant with nil side effects and can be prescribed as a drug to treat Parkinson's with subject to further trials.
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Eriocitrin alleviates the arterial occlusion-mediated cerebral ischemic-reperfusion injury through the modulation of apoptotic proteins and immune markers in mice p. 193

DOI:10.4103/pm.pm_577_19  
Background: Stroke is ranked on top of the persistent ailments associated with the brain. The incidence of ischemic stroke is rapidly rising along with intensified mortality as well as permanent disability conditions. However, tissue plasminogen activator (tissue-type plasminogen activator) is the only FDA-approved treatment for the treatment of stroke with decreased success rate. Eriocitrin, a flavonoid compound of Citrus limon with incredible therapeutic values, was active in the current investigation to explore the neuroprotective capability in arterial occlusion-provoked cerebral ischemic reperfusion injury (CIRI) in mouse replica. Materials and Methods: The neurological defects were considered through the standard scoring method and the expression status of apoptotic proteins, i.e., nuclear factor kappa B (NF-ƙB), caspase-3, and caspase-9, was examined via western blotting test. The expression of neuronal nitric oxide synthase (nNOS) was scrutinized through real-time polymerase chain reaction and standard ELISA kits were employed to examine the status of the proinflammatory markers. Results: The arterial occlusion provoked cerebral ischemic animals displayed the severely elevated status of inflammatory markers namely interleukin-6 (IL-6), IL-1β and tumor necrosis factor α, and the expression statuses of apoptotic proteins, as well as nNOS, were also uplifted while comparing to the sham subjected animals. Amazingly, eriocitrin (100 mg/kg) treatment revealed the declined inflammatory markers level, besides, the expression patterns of apoptotic proteins and nNOS were also reduced noticeably in the cerebral ischemic-reperfusion-operated animals. Conclusion: The novel findings of this investigation were demonstrated that the Eriocitrin delivered an astounding neuroprotection in the arterial occlusion-mediated CIRI in mouse replica, which possibly a promising neuroprotectant in cerebral ischemia.
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Protective effect of Viburnum grandiflorum against ultraviolet-B radiation-induced cellular and molecular changes in human epidermal keratinocytes p. 200
Hanjun Liu, Haixiu Zhang, Minyan Dang, Yukiat Lin, Hui Yan
DOI:10.4103/0973-1296.313853  
The aim of the present study was to evaluate the photoprotective potential of Viburnum grandiflorum (VG) against ultraviolet-B radiation-induced responses in HaCaT cells. The HaCaT cells were pretreated with VG prior ultraviolet-B (UVB)-radiation exposure and were further examined for lipid peroxidation, enzymatic antioxidant activity, % reactive oxygen species, DNA damage, mitochondrial membrane potential and for inflammatory, and apoptotic signaling markers such as tumor necrosis factor-alpha, nuclear factor kappa B, interleukin-1 (IL-1), IL-6, cyclooxygenase-2, p53, caspase-3/9, cytochrome-c, Bax, and Bcl-2. The VG pretreatment in UVB exposed cells shows significantly regulated both inflammatory as well as apoptotic signaling cascades. Our findings suggest that VG may be functional against UVB-induced photo-damages.
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RETRACTION Top

Retraction: Protective effect of viburnum grandiflorum against ultraviolet B radiation-induced cellular and molecular changes in human epidermal keratinocytes p. 207

DOI:10.4103/0973-1296.312818  
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ERRATUM Top

Erratum: Chemical composition and biological potential of a chloroform fraction from the leaves of marine plant syringodium filiforme kutzing p. 208

DOI:10.4103/0973-1296.313548  
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