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July-September 2020
Volume 12 | Issue 3
Page Nos. 207-335

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ORIGINAL ARTICLES  

Toxicological study of the effect in vivo and in vitro of Artemisia herba-alba aqueous extract in rats Highly accessed article p. 207
Ali Lahna, Naima Benjelloun, Nadia Seddik, Mernissi Farida, Abdallah Naya, Mounia Oudghiri
DOI:10.4103/pr.pr_4_20  
Background: Artemisia herba-alba (AHA) is largely used in folk medicine in different countries. However, rare studies provided toxicological evaluation regarding their safety on human health. Objective: This study investigated the safety of the standardized aqueous extract of AHA, like used by patients, to evaluate their toxicity in vivo and in vitro. Materials and Methods: For toxicological evaluation in vivo we used acute (during 14 days) and sub-acute oral gavages in Wistar rats (rats treated daily for 42 days at 1–5 g/kg bw) and the 3-[4, 5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay was performed to determine the level of cell viability and the degree of cytotoxicity in vitro (0–30 μg/ml) on cultured spleen cells. Results: The LD50was up to 2 g/kg. Signs of mortality and toxicity were observed after single doses and no-observed-adverse-effect levels in the sub acute toxicity was up to 2 g/kg bw. Compared to the control, the treatment did not produce any statistically significant changes on alanine aminotransferase and aspartate aminotransferase serum titer. However, for creatinine and urea serum value, a significant increase (P < 0.05) was observed. The histological observations of liver and spleen tissues have shown well-preserved normal cells. Indeed for kidney tissues some artifacts of retraction and vascular congestion were noted for 3–5 g/kg doses after sub-chronic treatment. The addition of plant extracts to the spleen cells did not show any sign of toxicity for all doses tested. Conclusion: We conclude that AHA aqueous extract at the dosage up to 2g/kg bw will be toxic and can affect mainly the kidney tissues.
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Liquid chromatography high-resolution mass spectrometry analysis, phytochemical and biological study of two aizoaceae plants: A new kaempferol derivative from Trianthema portulacastrum L. Highly accessed article p. 212
Hala Abuzaid, Elham Amin, Abeer Moawad, Usama Ramadan Abdelmohsen, Mona Hetta, Rabab Mohammed
DOI:10.4103/pr.pr_119_19  
Background: Natural remedies used for the treatment of liver diseases have a major concernworldwide. Objectives: Evaluation of the cytotoxic potential of successive fractions of two Aizoaceae plants; Trianthema portulacastrum L. and Aizoon canariense L. against human hepatocellular carcinoma (HepG2) cell lines. Moreover, metabolomic profiling of the successive fractions of both plants was carried out. Materials and Methods: Cytotoxic activity of successive fractions of the two plants against hepatocellular carcinoma (HCC) HepG2 cell lines were evaluated using viability assay, whereas metabolomic profiling was carried out using Liquid Chromatography High-Resolution ElectroSpray Ionization Mass Spectrometry (LC-HR-ESI-MS). Results: Significant cytotoxic activity of the n-hexane and n-butanol extracts of A. canariense (24.7 ± 3.5 and 55.3 ± 4.9 μg/mL, respectively) is recorded. On the other hand, metabolomic profiling of both plants resulted in dereplication of 27 metabolites belonging to different chemical classes; for example, sterols, flavonoids, diterpenes, triterpenes, tetraterpenes, alkaloids, lignans, hydrocarbons, and nucleosides. Phytochemical study of the biologically active fractions resulted in the isolation of one new compound; kaempferol-3-O-(2''-O-β-D-glucopyranosyl)-6''-O-E-feruloyl-β-D-glucopyranosid (T1). Biological testing of the isolated compounds indicated significant activity of T1 against HCC (IC50 =7.19 ± 0.27 μg/mL). Conclusion: Phytochemicals isolated from T. portulacastrum L and A. canariense L. may be responsible for their cytotoxic activity against HCC HepG2 cell lines.
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Terminalia chebula retz: A prospective agent in reducing the doxorubicin-mediated cardiotoxicity Highly accessed article p. 219
M Aamina, Ahmad Alhowail, Maha Aldubayan, Syed Imam Rabbani
DOI:10.4103/pr.pr_109_19  
Objectives: The present study evaluated the role of Terminalia chebula Retz. extract against doxorubicin (DXR)-mediated cardiac damages in male and female rats. Materials and Methods: The ethanolic extract of T. chebula (0.25, 0.5, and 1 g/kg) was administered simultaneously with DXR (2.5 mg/kg, three-doses on alternate days by intraperitoneal route). Two additional groups were evaluated by administering 0.5 g/kg of the extract to animals before or after DXR treatment. The study was done separately in male and female adult Wistar rats of bodyweight 180–200 mg. The cardiac biomarker levels such as creatine kinase-monoenzyme B, lactate dehydrogenase, alanine aminotransferase, and aspartate aminotransferase were estimated after each treatment. Histopathology of cardiac tissue was studies analyzed, and the level of superoxide dismutase was determined in serum. The results were statistically analyzed using one-way ANOVA and Bonferroni tests and P < 0.05 was considered to indicate the statistical significance. Results: The observations indicated that DXR significantly (P < 0.001) elevated the biomarker levels of cardiac damage in both male and female rats, besides inducing the structural changes in the myocardium tissues and antioxidant status. The co-administration of higher doses of T. chebula extract (0.5 and 1 g/kg) with DXR significantly (P < 0.01) reduced the cardiac enzyme levels and histopathological changes and improve the antioxidant status compared to DXR group. Post-treatment with T. chebula (0.5 mg/kg) showed mild inhibitory action on the DXR-induced cardiac changes without significantly affecting the antioxidant level. Conclusion: The results suggest that co-administration of T. chebula reduced the DXR-mediated cardiac damages and the action could be related to the enhancement of the antioxidant property in rats.
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Effect of a thai folk recipe on phytochemical screening, antioxidant activities, and α-Glucosidase inhibition by different solvent extracts p. 225
Anirut Namwong, Pramote Thongkrajai, Ampa Konsue
DOI:10.4103/pr.pr_100_19  
Context: Thai traditional medicine (TTM) has been widely use to treatment of various diseases. Aims: The aims of this study were determined on phytochemical screening, antioxidations, and α-glucosidase inhibition by different solvent extractions. Materials and Methods: The five medicinal plants from a TTM recipe were extracted using aqueous, 50% ethanol and 95% ethanol. The phytochemical screening were determined on total phenolic (TPC) and flavonoid (TFC) contents. Their antioxidant activities were tested using by 2,2-diphenyl-1-picrylhydrazy (DPPH) radical scavenging assay, 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS+) radical scavenging assay and ferric reducing antioxidant power (FRAP) assay. Glucose transferase mechanism was evaluated using by α-glucosidase inhibitory assay. Results: The results found that the recipe was ingredient with TPC and TFC. The aqueous extract (AE) (47.955 ± 0.256 mgGE/gExt) was significantly highest amount on TPC while, the ethanolic extract (EE) (1.988 ± 0.038 mgQE/gExt) was showed significantly highest amount on TFC. The antioxidations, DPPH, and the AE (IC50 =0.227 ± 0.005) were significantly more potent on free radical scavenging. ABTS+, AE (IC50 =0.148 ± 0.007), Hydro-ethanolic extract (HEE) (IC50 =0.161 ± 0.008), and EE (IC50 =0.151 ± 0.007) were not different on this method. FRAP, EE (12.681 ± 0.620 mgTE/gEt) were significantly more potent on cation radical reducing. The α-Glucosidase inhibitory activity, AE (IC50 =0.021 ± 0.002 mg/mL) and HEE (IC50 =0.076 ± 0.003 mg/mL) were significantly more potent on EE (IC50 =0.292 ± 0.010 mg/mL) and Acarbose® (IC50 =1.05 ± 0.110 mg/mL). Conclusion: The study confirms traditional use of a Thai folk herbal plants on antioxidation and α-glucosidase inhibition. The recipe was contained with also TPC and TFC might be more potential to antioxidant activities and anti-α-glucoxidase enzyme. Future study, we should be performed to clarify the mechanisms, major active compounds and in vivo.
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Antidiabetic and antidyslipidemic activities of hexane/ethyl acetate/methanol fractions of Paspalum scrobiculatum linn. grains in high-fat diet and streptozotocin-induced diabetic rats p. 230
G Jyothi Reddy, K Bhaskar Reddy, G V Subba Reddy
DOI:10.4103/pr.pr_6_20  
Background: Dietary habits and sedentary lifestyle are the major causes of increasing obesity, which in turn causes diabetes and cardiovascular problems. With this alarming increase, there is a need for alternative therapies to treat diabetes and hyperlipidemia. Objectives: To evaluate antidiabetic and antidyslipidemic effects of fractions of Paspalum scrobiculatum grains in rats. Materials and Methods: Different doses of Hexane: Ethyl acetate fraction of P. scrobiculatum (HEPS) and Ethyl acetate: Methanol fraction of P. scrobiculatum (EMPS) fractions of P. scrobiculatum were administered to the high-fat diet (HFD) and streptozotocin (STZ) (35 mg/kg)-induced diabetic rats with glibenclamide at 20 mg/kg body weight (b.w) as a standard reference. Biochemical and histopathological changes were assessed after 5 weeks of treatment. Results: Daily oral treatment with HEPS and EMPS each at 100 and 200 mg/kg b.w for 5 weeks was found to be significant in the reduction of plasma glucose levels, HbA1c, increased plasma insulin levels, and normalized lipid profile and liver function parameters in HFD and STZ-induced diabetic rats. In addition, degenerative histopathological changes seen in liver, kidney, and pancreas of diabetic rats were found to be normalized in HEPS and EMPS as comparable to that of glibenclamide-treated rats. Conclusion: The obtained results suggest that HEPS and EMPS fractions of P. scrobiculatum grains possess promising antihyperglycemic and antidyslipidemic potential without apparent toxic effects, which may prove the claimed use of the plant in treatment of diabetes and dyslipidemia.
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Effect of seasonal variation on euphadienol content in Euphorbia antiquorum L. and Euphorbia tirucalli L. using high-performance thin-layer chromatography method p. 236
Sudipta Roy, Rabinarayan Acharya, VJ Shukla, Anagha Ranade
DOI:10.4103/pr.pr_1_20  
Background: Ayurveda emphasizes that different parts of plants (both organized and unorganized) should be collected on a certain season to get their better yield and therapeutic value. In general, Kshira (latex) should be collected in Sharada rutu (autumn season). However, the latex of Snuhi has been stated to be collected at Shishiranta (end of winter). The latex of Snuhi has an important place in different therapeutic formulations. One of the important mentions is for the preparation of Ksharasutra, one of the widely used paramedical procedures in Ayurveda. Objectives: The objective of this study was to evaluate the impact of seasonal variation on the quantity of euphadienol content (one of the major active constituents) in latex sample of Euphorbia antiquorum L. (EA) and Euphorbia tirucalli L. (ET). Materials and Methods: The latex was collected individually during six different seasons in a calendar year from two species i.e. EA and ET. Then, after suitable sample preparation, it was subjected to high-performance thin-layer chromatography study using euphadienol as a standard biomarker and scanned at 210 nm. Results: The euphadienol content was found to be present in all the six seasons across the calendar year. The content was found maximum in Hemanta rutu and Sharada rutu samples in case of EA and ET, respectively. Conclusion: The study proves that changes of seasons do have a certain effect on the phytoconstituents in the plants.
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A phytochemical analysis and in vivo effects of an herbal aphrodisiac Newtonia hildebrandtii on male wistar rat reproductive system p. 243
Thomson Msiska, Anthony Mwakikunga, David Tembo, Fanuel Lampiao
DOI:10.4103/pr.pr_112_19  
Background: The study was aimed at screening phytochemicals and evaluating the effects of aqueous and 50% (v/v) methanol stem bark extracts of Newtonia hildebrandtii on male rat reproductive system. Materials and Methods: Thirty male Wistar rats were divided into five groups and were given different treatments of N. hildebrandtii extracts at 200 and 400 mg/kg body weight. After 8 weeks, body and right testis weight, sperm concentration, hormonal levels, and histological structure of the testis were evaluated. Calorimetric method was used for quantitative analysis of phenolic compounds, whereas gas chromatography–mass spectrometry (GC-MS) was used for fingerprinting of chemical compounds. Results: Testosterone, luteinizing hormone, estradiol, and follicle-stimulating hormone levels were significantly increased, whereas prolactin levels were significantly decreased in the treated groups compared to the control. Sperm parameters were significantly increased compared to the control. Treated animals showed significant increases in body weight, seminiferous tubule diameter, and germinal epithelial height with the testes showing all the stages of spermatogenesis. Phytochemical screening of the methanolic extracts revealed the presence of alkaloids, carbohydrates, reducing sugars, steroids, anthraquinones, flavonoids, phenols, and saponins. GC-MS analysis revealed the presence of dimethylsilanediol; (R, R)-2,3-butanediol; hexamethylcyclotrisiloxane; succinimide; 1H-1,2,3-triazole-4-carboxylic acid, 1-(4-amino-1,2,5-oxadiazol-3-yl)-5-ethyl-, ethyl ester; l-(+)-ascorbic acid 2,6-dihexadecanoate; cis-13,16 docasadienoic acid, methyl ester; 9-octadecenoic acid (Z)-, 2-hydroxy-1-(hydroxymethyl) ethyl ester; and 2-chloro-benzoic acid N'-[2-(3-oxo-3,4-dihydro-2H-benzo[1,4]thiazin-2-yl)-acetyl]-hydrazide. Conclusion: N. hildebrandtii appears to possess androgenic and anabolic effects. The aforementioned phytochemicals seem to be responsible for its aphrodisiac effects. This study supports the ethno-aphrodisiac claims that have been made against it.
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Kaempferol – A dietary flavonoid isolated from blepharis integrifolia p. 250
KM Anil Kumar, Ramith Ramu, S Chandan, Shivaraju Harikaranahalli Puttaiah
DOI:10.4103/pr.pr_114_19  
Background: Blepharis integrifolia is in the preparation of traditional formulation for the treatment of various diseases, including digestive disorders such as flatulence and dysentery. Although a very few studies have assessed the pharmacological properties, nothing is known about the phytochemicals present and the effect of extracts on its pharmacological activities. Objectives: Therefore, the objectives of the present study were to assess the role of methanol extract of B. integrifolia (MBI) for its anti-inflammatory, antioxidant, and antibacterial activities. Materials and Methods: The phytochemicals present in the extract were evaluated using high-resolution liquid chromatography and mass spectrometry (HR-LCMS). The extract and the isolated compound were subjected to antioxidant, antibacterial, and anti-inflammatory studies. Results: MBI exhibited an effective antibacterial, antioxidant, and anti-inflammatory activities at a more significant level than the standards. HR-LCMS analysis of MBI revealed the presence of kaempferol, rutin, and several phytochemicals. The activity-guided repeated fractionation of the methanol extract by silica gel column chromatography yielded a compound that exhibited strong antioxidant activity. Based on various physicochemical and spectroscopic analyses (ultraviolet, infrared, proton nuclear magnetic resonance [NMR], carbon-13 NMR, and MS), the bioactive compound isolated was elucidated as 3,5,7-trihydroxy-2-(4-hydroxyphenyl)-4H-chromen-4-one (kaempferol). Furthermore, the molecular docking studies for kaempferol revealed that it binds β-lactamase at MEG binding site and was found to interact with binding site residues PHE91, TRP94, ASN109, MET148, GLU147, ASN36, and ILE201. Conclusion: The results from the present study suggest that the potent antioxidant, antibacterial, and anti-inflammatory activities observed are a result of the presence of these bioactive compounds within the extract. In addition, these results also demonstrate the antioxidant potency of kaempferol which could be the basis for its alleged health-promoting potential.
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Screening of cytotoxic activity of hematite (α-Fe2O3) nanoparticles from Butea monosperma on MCF-7 cells p. 260
Debasish Pradhan, Shaktiprasad Pradhan, Bandana Behera, Adyasa Samantaray
DOI:10.4103/pr.pr_115_19  
Background: Hematite (α-Fe2O3) has been used as an antimicrobial and disinfectant agent. Nevertheless, there are limited data about antitumor potential. This study was focused on investigating cytotoxic effects of Hematite (α-Fe2O3) from Butea monosperma flower extract on MCF -7 breast cancer cells and its mechanism of action. Materials and Methods: Thus, a green method was created for the synthesis of Hematite (α-Fe2O3) using an aqueous extract of B. monosperma flower. Synthesis of Hematite (α-Fe2O3) was described by different analytical techniques including ultraviolet-visible spectrophotometer, field-emission scanning electron microscopy, X-ray diffraction, and Fourier transforms infrared spectroscopy. Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Reactive oxygen species (ROS) formation was measured using probe 2',7'-dichlorofluorescein diacetate and intracellular calcium (Cai2+) was evaluated with probe flu3-AM. Cells were treated with different concentrations of Hematite (α-Fe2O3) (1, 3, 6, 10, 15, 25, 50, and 100 μg/mL). Results: The results showed that Hematite (α-Fe2O3) hindered cell growth in a dose-dependent manner. Hematite (α-Fe2O3) appeared to have dose-dependent cytotoxicity against MCF-7 cells through activation of the ROS generation and an increase in the intracellular Cai2+ (half-maximal inhibitory concentration 52 ± 3.14). Conclusion: In conclusion, the results of this preliminary study demonstrated that Hematite (α-Fe2O3) from B. monosperma flower extract may be a potential therapeutic potential medicament for human breast cancer treatment.
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Evaluation of cytotoxicity and antioxidant potential of bael leaf (Aegle marmelos) on human hepatocellular carcinoma cell line p. 267
Kanokwan Kulprachakarn, Sakaewan Ounjaijean, Somdet Srichairatanakool, Duangta Kanjanapothi
DOI:10.4103/pr.pr_15_20  
Background: Aegle marmelos (Bael) is one of the important medicinal plants for curing human diseases. Nevertheless, the information about the cytotoxic and antioxidant effects of Bael leaf is not much. Objectives: The objective of this study is to evaluate the cytotoxicity and antioxidant activity of Bael leaf in human hepatocellular carcinoma (HepG2) cell lines. Materials and Methods: Bael leaves extract was prepared by 80% (v/v) aqueous ethanol and measured for total phenolic compound and antioxidant capacity. The ability of ethanolic Bael leaf extract to inhibit hepatic cancer cells (HepG2 cell) growth was evaluated by determining the percentages of cell viability. The levels of intracellular reactive-oxygen species (ROS) and lipid peroxidation were determined. Results: Bael leaf extract demonstrated the potentiality to inhibit the cancer cell growth with a 50% inhibitory concentration (IC50) at 50 μg/ml and 72 μg/ml after 24 h and 48 h incubated times, respectively. The total phenolic content and antioxidant capacity of ethanolic extract were 218.33 ± 43.81 mg gallic acid equivalent/g of dry extract and 33.62 ± 3.07 mg Trolox equivalent antioxidant capacity/g of dry extract, respectively. At the lowest concentration of Bael leaf extract, it was found that Bael leaf extract shows the antioxidant activity by the ability to lowering the level of ROS and malondialdehyde in HepG2 cell. Conclusion: Bael leaf has a high antioxidant component, which is beneficial and can be developed as new therapeutic uses. However, further studies on the benefits of Bael leaf should be performed for better realizing and effective use soon.
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Chemical analysis and biological potentials of extracts from Colpomenia sinuosa p. 272
Reem Monal Al Monla, Zeina T Dassouki, Hala Gali-Muhtasib, Hiba R Mawlawi
DOI:10.4103/pr.pr_91_19  
Background: Colpomenia sinuosa is a brown alga rich in molecules of pharmacological, nutraceutical, and functional properties. Despite its therapeutic potentials, this alga is poorly studied in this area of the Mediterranean. Objectives: The study objective was to investigate the physicochemical properties and biological potentials of different extracts of the brown seaweed C. sinuosa. Materials and Methods: Crude extracts and fractions were obtained by different types of solvent extractions. Proximal analysis was adopted to reveal the ash content. Protein and carbohydrate contents were determined by phenol-sulfuric acid method and Lowry and Bradford methods, respectively. The mineral content was assessed by atomic absorption spectroscopy, where the total phenolic content (TPC) was determined by Folin–Ciocalteu reagent, antioxidant properties by radical scavenging assay, and antimicrobial activity by disc diffusion method. Finally, the anti-inflammatory activity was determined by albumin denaturation test. Results: Proximal analysis revealed high levels of total ash content (20.03 ± 0.79%), protein content (10.77 ± 0.18%), total lipids (4.35 ± 0.212%), and total carbohydrates (40.367 ± 0.421%). Magnesium concentration (16.79 ± 0.721 mg/g) was highest among macro minerals. The TPC of fractions and extracts ranged from 17.78 ± 0.092 to 39.02 ± 0.199 mg gallic acid equivalents/g dry weight. The highest scavenging activity reached 84.1% ± 0.83 at 750 μg/mL. Maximal anti-inflammatory activity was noted in aqueous extract (59.38 ± 4.35 μg/mL). Conclusion: This significant scavenging activity increased in polar extracts obtained by Soxhlet extraction. The aqueous extracts showed the highest 2,2-diphenyl-1-picrylhydrazyl scavenging activity and highest albumin denaturation inhibition, whereas the highest bactericidal activity was found in the methanolic extracts. Our results represent the first report on Lebanese C. sinuosa extract as a promising source of bioactive compounds with high antioxidant potential.
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Effect of carbon tetrachloride-induced hepatic injury on Luffa acutangula (var.) amara p. 278
E Prabhu Daniel, AR Vijayakumar, W Clement Atlee, R Ilavarasan, M Sivakumar
DOI:10.4103/pr.pr_113_19  
Objectives: To study the reversal effect of carbon tetrachloride (CCl4)-induced hepatic injury on the ethyl acetate and ethanol extracts of Luffa acutangula var. amara in Wistar rats. Materials and Methods: The study included experimental mice groups in which pentobarbital-induced sleeping time and its prolongation by CCl4is assessed by righting reflex and recovery time. Chronic liver injury was induced by oral administration of CCl4and acute injury by a single intraperitoneal injection in experimental Wistar rats. Blood and liver specimens were used for analysis of enzymatic reactions. Results: It was noted that the mice groups treated with extracts decrease the sleeping time in and standard silymarin than the control group. Acute and chronic liver injury models showed increases in total protein and glutathione peroxidase, whereas there was a reduction in total bilirubin and serum alkaline phosphate level in extracts and standard silymarin-treated rats than control. Histopathological studies showed a significant reduction in inflammation, fatty change and gross necrosis in extracts and standard silymarin-treated animals than control. Conclusion: The study suggests that extracts of L. acutangula var. amara can be useful in treating acute and chronic hepatic injury.
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Comparative botanical and phytochemical studies of ambiguous medicinal plant species of Wedelia and Eclipta (Fam. Asteraceae) used in ASU systems of medicine with special reference to in-silico screening of hepatoprotective potential of marker wedelolactone with acetaminophen targets p. 285
C Arunachalam, R Arunadevi, S Murugammal, N Monika, R Susila, S N Sunil Kumar
DOI:10.4103/pr.pr_11_20  
Context: In traditional medicine, Kesaraja (Ayurveda) or Manjal karisali (Siddha) is effective for jaundice. Aim: Three species of Asteraceae need to be studied for their therapeutic superiority of their intended claim. They are Wedelia chinensis (Osbeck) Merr. Philipp J., Wedelia trilobata (L.) Hitchc. and Eclipta prostrata (L.) L. (Asteraceae). The present study aimed to screen and characterize the potential species for therapeutic purpose. Materials and Methods: The whole plants, W. chinensis (Osbeck) Merr. Philipp J., W. trilobata (L.) Hitchc. and Eclipta prostrata (L.) (Asteraceae) were collected and botanically identified. Preliminary phytochemical analysis and high-performance thin-layer chromatography finger printing with marker wedelolactone were done for the ethanolic extracts of these plants. Botanical and pharmacognostical diagnostic characters of the plants based on macro-morphological, micro-morphological and powder microscopical characterization were worked out. Comparative in-vitro antioxidant potential of ethanolic extracts of these plant species was carried out. Using ADMET SAR software, the pharmacokinetics of wedelolactone were predicted. Using Autodock 4.2 software, the binding energy of wedelolactone on targets of acetaminophen-induced hepatotoxicity namely PPAR-α, AMPK, JNK-1, EGFR, Nrf2, ALT, ALP, GGT, CAR, Frizzled receptor, FXR, ERK1, LXR, mitochondrial glutamate dehydrogenase, p53, mTOR C1, CYP1A2, CYP2E1, 5-lipoxygenase, thrombin, UCP1, GSK1, RXR and PXR was predicted. Results: All the three plant species were pharmacognostically and chemically different. W. chinensis was found to possess more antioxidant potential than the other two plants. The marker compound wedelolactone was not detected in W. trilobata. Wedelolactone passed the Lipinski's rule of five, and the docking analysis of wedelolactone confirmed high binding affinity toward PPAR-α, AMPK, Nrf2, CYP2E1, EGFR, JNK1, UCP-2, thrombin, 5-lipoxygenase, mTORC1, RXR, FXR, LXR, Frizzled receptor, GDH and Erk-1. Conclusion: Based on the above observations, we conclude that the presence of marker compound wedelolactone might have attributed the potency of W. chinensis and E. prostrata in counteracting acetaminophen toxicity when compared with W. trilobata.
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Citrus limon L. gel stimulate lymphocytes and interleukin-10 expression in traumatic diabetic oral ulcers p. 299
Fatma Yasmin Mahdani, Diah Savitri Ernawati, Priyo Hadi, Bagus Soebadi, Shiffin Devi Mardiyana, Dwi Susanti, Meircurius Dwi Condro Surboyo
DOI:10.4103/pr.pr_12_20  
Background: Citrus limon L. and its peel report anti-inflammatory, antioxidant, anti-fungal, and anti-bacterial properties. However, the effects of C. limon L. gel to treat traumatic oral ulcers on diabetic conditions have yet to be investigated. Purpose: This study aims to discover the potent effect of C. limon L. on traumatic diabetic oral ulcers, in terms of both lymphocytes and interleukin (IL)-10 expression. Materials and Methods: C. limon L. gel was made from C. limon L. peel and was extracted by means of steam distillation. Twenty male, diabetic Wistar rats with traumatic ulcers on the inferior labial mucosa were divided into four groups (n = 5). The control groups were treated with 3% carboxylmethyl celullosa natrium and the treatment groups were treated with C. limon L. gel. Each treatment was administered once a day for either 5 or 7 days. Lymphocytes were analyzed by means of hematoxylin and eosin staining and IL-10 expression by immunohistochemistry staining. An independent t-test (P < 0.05) was used to analyze and determine the number of lymphocytes and IL-10 expression. Results: The number of lymphocytes and IL-10 expression in the treatment groups on the fifth and 7th days was higher than those in the control groups (P < 0.05). Conclusion: The essential oil in C. limon L. peel can be used to treat traumatic diabetic oral ulcers, as it is able to stimulate the number of lymphocytes and IL-10 expression.
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Cholinergic-enhancing and antioxidant effect of Vigna subterranea (L.) Verdc. (Fabaceae) landrace aqueous extract on scopolamine-induced amnesia in male swiss mice p. 303
Hervé Hervé Ngatanko Abaissou, Harquin Simplice Foyet, Eglantine Keugong Wado, Balbine Kamleu Nkwingwa, Jorelle Linda Damo Kamda, Lea Blondelle Kenko Djomessie, Alin Ciobica, Elisabeth Ngo Bum
DOI:10.4103/pr.pr_62_19  
Background: Currently, no drug effectively curbs down the progression of Alzheimer's disease (AD), and thus early prevention is important. Objectives: The present study was conducted to investigate the protective effects of the aqueous extract of Vigna subterranea seed landrace on amnesia induced by scopolamine in mice. Materials and Methods: V. subterranea aqueous extract (100, 200, and 400 mg/kg BW) was administered by gavage for 9 consecutive days and memory impairment was induced by repeated intraperitoneal injection of scopolamine (1.5 mg/kg). The Y-maze (YM), Morris water maze (MWM), novel object recognition paradigm (NOR), and the T maze™ were used to assess learning, memory, and retention. Superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) levels, and acetyclcholine esterase activity were also evaluated in the mouse hippocampi homogenates. Results: V. subterranea aqueous extract (400 mg/kg) significantly increased the percentage of spontaneous alternation in the YM task and decreased escape latency in the MWM. Moreover, this dose brought about a significant improvement in the time spent in the preferred TM arm and discrimination index in the NOR tasks despite repeated scopolamine injection. Additionally, low acetylcholine esterase levels, reduced lipid peroxidation (MDA), but increased antioxidant enzyme (CAT and SOD) activity were observed in hippocampi homogenate of mice pretreated with the extract. A protective action against hippocampal cell damage was also evident. Conclusion: This finding suggests that the aqueous extract of V. subterranea seed landrace may improve learning and memory.
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Determination of flavan-3-ols by high-performance liquid chromatography-photodiode array, antioxidant potential, antimicrobial activity, and pharmacognostic evaluation of Maytenus obtusifolia (Mart.) leaves p. 313
Diego Igor Alves Fernandes de Araújo, Anny Palloma de Lima Arruda Fernandes, Anderson Angel Vieira Pinheiro, Augusto Lopes Souto, Kaio Aragão Sales, Priscylla Rayama Alves Fernandes de Araújo, Rodrigo Silva de Andrade, Marcelo Sobral da Silva
DOI:10.4103/pr.pr_9_20  
Background: Maytenus obtusifolia Mart. (Syn. Monteverdia obtusifolia Mart. Biral), Celastraceae, is a species located on the coast of Brazil, which is commonly used as anti-inflammatory and on the treatment of gastric ulcers. Objectives: The aim of this study consists of the determination of quality control parameters of the plant material, as well as to develop, validate high-performance liquid chromatography-photodiode array methodology for the quantification of its analytical markers and evaluate the antioxidant potential and antimicrobial activity of the extract. Materials and Methods: The plant material and ethanolic crude extract (ECE) were evaluated through pharmacopoeial and non-pharmacopoeial methods. Anti-oxidant assay was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Minimum inhibitory concentration was used to evaluate antimicrobial activity assay. Results: The assays have provided the following results: granulometry (d50 0.82 mm), moisture content (6.41%–9.03%), pH of aqueous solution (5.41), apparent bulk density (0.41 mg/mL), heavy metals (<10 ppm), total ash (5.20%), and sulfated ash content (6.36%). Total flavonoid (2.14%; 6.24%), tannin (6.51%; 17.85%), and phenolic content (41.95; 134.76 mg AG/g) regarding leaves powder and its ECE, respectively. The validated method was specific, linear, precise, accurate, and robust. The content of (-)-Epigallocatechin, (+)-Catechin and (-)-Epicatechin on the ECE was 1.00%, 0.40%, and 0.33%, respectively. The anti-oxidant activity was evaluated by DPPH assay and have presented an IC50at 18.01 μg/mL. In addition, the most important antimicrobial activity of the ECE was against Aspergillus flavus (MIC at 128 μg/mL). Conclusion: These data have demonstrated to be very useful toward the analysis and pharmacognostic quality control of M. obtusifolia Mart.
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Trypanocidal potentialities of skimmianine an alkaloid isolated from Zanthoxylum pistaciifolium griseb leaves p. 322
Yamilé Heredia Diaz, Julio César Escalona Arranz, Rosalia González Fernández, Ania Ochoa Pacheco, Jesus García Díaz, Lianet Monzote Fidalgo, Denise da Gama Jaén Batista, Cristiane Franca da Silva, Paul Cos
DOI:10.4103/pr.pr_44_19  
Background: Zanthoxylum pistaciifolium Griseb (Rutaceae) is an endemic Cuban species traditionally used for respiratory ailment treatments and religious practices. Objective: The aim of this study was to evaluate the in vitro antimicrobial activity from Z. pistaciifolium leaves. Materials and Methods: Four phases obtained from the ethanolic extract were tested against bacteria (Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa), fungi (six Candida spp.) and parasites (Leishmania amazonensis and Trypanosoma cruzi) by microdilution methods. In addition, the cytotoxicity was determined against peritoneal macrophages from BALB/c mice and fibroblast cells L929 using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and resazurin assay. Results: The hexane-butanone phase was the most active against Candida spp. and exhibited moderate effect on T. cruzi trypomastigote (39.3 ± 5.5 μg/mL). The skimmianine isolated from this phase and characterized by nuclear magnetic resonance showed a median inhibitory concentration of 58.0 ± 4.0 μg/mL against T. cruzi trypomastigotes and 31.2% ± 6.5% of inhibition on amastigote form at 100 μg/mL. Antiprotozoal activity of this alkaloid was selective with a selectivity index of 9. Conclusion: The current study demonstrated for the first time the antimicrobial potential of the leaves of Z. pistaciifolium, supporting its ethnobotanical uses. The trypanocidal activity observed for the alkaloid skimmianine could contribute to development future of an herbal medicine for the treatment of Chagas diseases.
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Evaluation of the antioxidant and gastric antiulcerogenic activities of the hydroalcoholic extract and leaf fractions of Solanum stipulaceum roem. and schult. p. 328
Cirlane Alves Araujo de Lima, Daniela Feitosa de Oliveira, Andressa da Silva, Robson Silva de Lima, Jymmys Lopes dos Santos, Antônio Santos Dias, André Luiz Lima Menezes dos Santos, Jamylle Nunes de Souza Ferro, Andrea Yu Kwan Villar Shan, Brancilene Santos de Araujo, Emiliano de Oliveira Barreto, Maria Silene da Silva, Josemar Sena Batista, Sara Maria Thomazzi, Sonia Piacente, Cosimo Pizza, Charles dos Santos Estevam
DOI:10.4103/pr.pr_2_20  
Background: An earlier ethnobotanical study showed that the species Solanum stipulaceum was indicated first in many orders for the treatment of gastric ulcers. Aims: The aim of this study was to evaluate phytochemical profile and the antioxidant, antiulcerogenic, protective redox, and cytotoxicity activities of the hydroethanol extract (HEE) and leaf fractions of this plant. Materials and Methods: The HEE was obtained by maceration and fractionated by liquid–liquid extraction in fractions: hexane fraction, chloroform fraction (CF), ethyl acetate fraction (EAF), and hydromethanol fraction (HMF). Phytochemical profile was evaluated by colorimetric methods. Antioxidant activity was measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The cellular toxicity was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In vivo antiulcerogenic activity was investigated using the ethanol-induced acute ulcer model. Afterward, protective redox activity was assessed with the plasma and liver of these animals using the thiobarbituric acid reactive substances assay. Results: Tannins, free steroids, triterpenes, alkaloids, and flavonoids were found in HEE, and total flavonoid measured was 5.26 ± 0.23 μg/mg of the extract. The DPPH activity was higher than 70% for HEE, EAF, and CF. Treatment with HEE (200 and 400 mg/kg) and CF, EAF, and HMF (200 mg/kg) inhibited the ulcer. CF reduces gastric lesions by 88.29% (50 mg/kg) and 85.65% (100 mg/kg). The HEE (400 mg/kg), CF, and EAF (200 mg/kg) reduced oxidative stress in the plasma and in the liver. HMF (200 mg/kg) reduced oxidative stress only in the liver. The HEE and fractions 10–50 μg/mL (P < 0.05) were nontoxic, maintaining cell viability at 65%. Conclusion: The extract and fractions have antioxidant activity in vitro and ex vivo, antiulcerogenic effect in vivo, and low cytotoxicity up to 50 μg/mL.
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