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   2017| October-December  | Volume 9 | Issue 4  
    Online since November 16, 2017

 
 
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ORIGINAL ARTICLES
Identification of the adulterated Asini Corii Colla with cytochrome c oxidase subunit I gene-based polymerase chain reaction
Hua-Li Zuo, Jie Zhao, Yi-Tao Wang, Zhi-Ning Xia, Yuan-Jia Hu, Feng-Qing Yang
October-December 2017, 9(4):313-318
DOI:10.4103/pr.pr_33_17  PMID:29263623
Background: Asini Corii Colla (ACC) (namely donkey hide gelatin, E'jiao in Chinese) was one of the most valuable tonic traditional Chinese medicines which is an infallible remedy to promote hematopoiesis. It should be produced by fresh or dried donkey hide according to Chinese Pharmacopeia (2015 edition) with a long-time decoction, while as donkey and horse (or mule) all belong to equids so their hides or their hide gelatins are share much in common, that cause the difficult in distinguishing raw materials donkey hide from horse/mule hide for manufacturer, and the challenge in the quality evaluation of ACC for regulatory authority to identify the adulterated with horse hide. Objective: To establish an effective quality evaluation methods for ACC focused on the qualitative-based identification of the raw material's authenticity, mainly to identify the species origin of the gelatins. Materials and Methods: DNA extracted from (1) Raw materials (hides of donkey, horse, mule, bovine and pig); (2) Five hide-glues (bovine, pig, donkey, horse and mule hide-glue); (3) 11 batches of ACC commercial products made by different manufactures from local drug stores. Polymerase chain reaction (PCR) method with newly designed horse-specific primers I and primer pair II. Results: Use the primer pair I, a 234 bp target product could be amplified sensitively from the DNA sample of horse/mule adulterated commercial ACC products, though the DNA in commercial products is severely degraded. A 219 bp product could be amplified specifically from the DNA sample of horse/mule hide, while the results were all negative for the DNA templates of donkey hide, its gelatin and ACC products without adulteration. Conclusion: The developed PCR method based on primer I and II provide an effective approach to identify the species origin of highly processed product ACC (primer pair I) as well as to distinguish the raw material donkey hide (primer pair II), which might enlighten a new strategy to the Quality Evaluation of ACC. Abbreviations Used: ACC: Asini Corii Colla; TCMs: Traditional Chinese Medicines; SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis; IEF: Isoelectric focusing; GFC: Gel filtration chromatography; 2-DE: Two-dimensional electrophoresis; PCR: Polymerase chain reaction
  5,279 31 -
Medicinal plants used by traditional healers in Sangurur, Elgeyo Marakwet County, Kenya
Gabriel Kigen, Wilson Kipkore, Bernard Wanjohi, Boniface Haruki, Jemutai Kemboi
October-December 2017, 9(4):333-347
DOI:10.4103/pr.pr_42_17  PMID:29263626
Background: Although herbal medical products are still widely used in Kenya, many of the medicinal plants used by traditional medical practitioners (TMPs) have not been documented, despite several challenges that are now threatening the sustainability of the practice. Objective: To document the medicinal plants and healing methods used by TMPs in a region of Kenya with several recognized herbalists for potential research. Materials and Methods: Semi-structured interviews, group discussions, and direct observations were used to collect ethnopharmacological information. The participant's bio-data, clinical conditions treated, methods of treatment, medicinal plants used, methods of preparation and administration, and dosage forms were recorded. Results: A total of 99 medicinal plants and 12 complementary preparations employed in the treatment of 64 medical conditions were identified. The most widely used plant was Rotala tenella which was used to treat nine medicinal conditions; seven each for Aloe tweediae and Dovyalis abyssinica; and six each for Basella alba and Euclea divinorum. The plants belonged to 55 families with Fabaceae family being the most frequently used (10), followed by Apocynaceae and Solanaceae, each with six species, respectively. We identified plants used to determine the sex of an unborn baby and those used to treat several conditions including anthrax and cerebral malaria and herbs used to detoxify meat from an animal that has died from anthrax. Of special interest was R. tenella which is used to prevent muscle injury. Conclusions: We have documented several plants with potential therapeutic effects. Further research may be conducted to determine their efficacy. Abbreviations Used: Fic: Informant consensus factor, Nur: Number of use reports in each category, Ns: Number of reported species, TMPs: Traditional medical practitioners.
  3,513 49 -
A standardized composition comprised of extracts from Rosmarinus officinalis, Annona squamosa and Zanthoxylum clava-herculis for cellulite
Mesfin Yimam, Young-Chul Lee, Ping Jiao, Mei Hong, Lidia Brownell, Qi Jia
October-December 2017, 9(4):319-324
DOI:10.4103/pr.pr_70_17  PMID:29263624
Background: Cellulite, characterized by changes in the skin morphology presented as dimpled or puckered skin appearance, is highly prevalent among postadolescent women. Cellulite management ranges from topical cream applications to invasive procedures. While some interventions showed improvements in physical appearances of affected areas, so far, none have reversed the condition to a full recovery. These unsuccessful measures signify the intricate nature of cellulite etiology highlighting its complexity leading to the possibility for a combination treatment approach to target multiple mechanisms. Materials and Methods: We screened our plant library for extracts that reduce cellular lipid accumulation, improve microcirculation, possess high total antioxidant capacity, significant anti-platelet aggregation, and anti-inflammatory activities using lipid accumulation assay in 3T3-L1 cells, Croton oil-induced hemorrhoid test in rats as a model for microcirculation, anti-platelet aggregation assay, nitric oxide (NO) inhibition assay, and 1,1-diphenyl-2-picrylhydrazyl assay. Results: Three known botanicals such as Rosemary officinalis, Annona squamosa and Zanthoxylum clava-herculis were identified as lead extracts in these tests. Treatment of 3T3 cell with A. squamosa at 1 μg/ml resulted in 68.8% reduction in lipid accumulation. In croton oil-induced hemorrhoid study, Z. clava-herculis reduced the recto-anus coefficient by 79.6% at 6 mg/kg indicating improvement in microcirculations. Similarly, R. officinalis caused inhibition of 82%, 71.8%, and 91.8% in platelet aggregation, NO production and free radical generation at 31.25 μg/ml, 6.2 μg/ml, and 40 μg/ml concentrations suggesting its anti-oxidant, and anti-inflammatory activities. Conclusions: Data depicted here suggest that formulation of these well-known botanicals at a specific ratio perhaps may yield a composition with a much wider spectrum of mechanisms of actions to impact the multiple pathways involved in cellulite onset, continuation, or exacerbations. Abbreviations Used: GMP: Good Manufacturing Practice; CA: Carnosic acid; NF-kB: nuclear factor-kB; HPLC: high-performance liquid chromatography; EtOH: Ethanol; DMEM: Dulbecco's modified Eagle's medium; FBS: fetal bovine serum; SD: Sprague Dawley; RAC: recto-anus coefficient; LPS: Lipopolysaccharide; DPPH: 1,1-diphenyl-2-picryl-hydrazyl; TNF-α: tumor necrosis factor; NO: Nitric oxide
  2,648 42 -
Ethnopharmacological survey of medicinal plants in Albaha Region, Saudi Arabia
Nasser A Awadh Ali, Saeed Salah Al Sokari, Ahmed Gushash, Sirajudheen Anwar, Khalid Al-Karani, Abdulwali Al-Khulaidi
October-December 2017, 9(4):401-407
DOI:10.4103/pr.pr_11_17  PMID:29263636
Background: Local natural medicinal resource knowledge is important to define and elaborate usage of herbs, in systematic and organized manner. Until recently, there has been little scientifically written document regarding the traditional uses of medicinal plants in Al Bahah region. Objective: This pilot study aims to collect the ethnobotanical information from native populations regarding the benefits of medicinal plants of Al Bahah region, and determine if the traditional usage is scientifically established (proved) from literature. Materials and Methods: The survey collected data for 39 plant species recorded by informants for their medicinal benefits. The recorded species were distributed among 28 plant families. Leguminosae and Euphorbiaceae were represented each by 3 species, followed by Asteraceae (2 species), Lamiaceae (2 species), Apocynaceae (2 species), and Solanaceae (2 species). All the medicinal plants were reported in their local names. Analysis of ethnopharmacological data was done to obtain percentage of plant families, species, parts of plants used, mode of administration, and preparation types. Results: Total 43 informants were interviewed, maximum number of species were used to cure skin diseases including burns (3), wounds (7), warts (1), Leishmania (7), topical hemostatic (2), followed by gastrointestinal system, rheumatism, respiratory tract problems, diabetes mellitus, anti-snake venom, malaria, and eye inflammation. Conclusions: The study covered Al Bahah city and its outskirts. Ten new ethnobotanical uses were recorded such as antirheumatic and anti-vitiligo uses for Clematis hirsute, leishmaniasis use of Commiphora gileadensis, antigout of Juniperus procera, removing warts for Ficus palmata. Abbreviations Used: UI: Use Index, GI: Gastrointestinal tract, RD: Rheumatic disease, CVS: Cardiovascular diseases, UTI: Urinary tract infection, DM: Diabetes mellitus, RT: Respiratory infection, KSA: Kingdom of Saudi Arabia
  2,443 42 -
Protective effect of Withania coagulans fruit extract on cisplatin-induced nephrotoxicity in rats
Sonam Sharma, Apurva Joshi, Siva Hemalatha
October-December 2017, 9(4):354-361
DOI:10.4103/pr.pr_1_17  PMID:29263628
Background: Fruits of Withania coagulans (Solanaceae) reported to possess several bioactive compounds as curative agents for various clinical conditions. Cisplatin is a chemotherapeutic drug to treat sarcomas, carcinomas, lymphomas, cervical cancer, germ cell tumors, etc. The major factor that limits its clinical use is its dose-dependent nephrotoxicity. Aim: To explore the nephroprotective effect of W. coagulans extract and its modulatory effects against cisplatin-induced nephrotoxicity and genotoxicity. Materials and Methods: W. coagulans fruit extract was quantitatively standardized with withaferin A using high-performance thin-layer chromatography. The subacute toxicity study was performed according to OECD guidelines in experimental rats. Nephrotoxicity in rats was induced by a single dose of cisplatin (6 mg/kg, intraperitoneal). Nephroprotective role of W. coagulans fruit extract at different doses had been evaluated. It includes quantification of serum kidney toxicity markers, renal tissue oxidative stress biomarkers and pro-inflammatory cytokines level, DNA fragmentation assay, and histopathological examination of renal tissue. Results: Withaferin A was found 3.56 mg/g of W. coagulans fruit extract. It significantly prevented the rise in serum urea and creatinine level and also preserve rat kidneys from oxidative stress and free radical induced DNA damage. Histopathological study showed extract treatment eliminates tubular swelling, cellular necrosis, and protein cast deposition in cisplatin treated kidney tissue. It averted the decline in glutathione content, activities of superoxide dismutase and catalase. These parameters were restored to near normal levels by extract in a dose of 400 mg/kg, per oral. Conclusion: It can be justified that W. coagulans possess dose dependent protective effect against cisplatin induced kidney damages, primarily through its free radical scavenging and anti inflammatory activity Abbreviations Used: WHO: World Health Organization, SOD: Superoxide dismutase, CAT: Catalase, HPTLC: High-performance thin layer chromatography, p.o.: Per-oral, i.p.: Intraperitoneal, TNF-α: Tumor necrosis factor-alpha, IL-1β: Interleukin 1-beta, IL-6: Interleukin-6
  2,274 33 -
REVIEW ARTICLE
A review of botany and pharmacological effect and chemical composition of Echinophora species growing in Iran
Zohreh Hosseini, Zahra Lorigooini, Mahmoud Rafieian-Kopaei, Hamzeh Ali Shirmardi, Kamal Solati
October-December 2017, 9(4):305-312
DOI:10.4103/pr.pr_22_17  PMID:29263622
This review was conducted to investigate the botany, phytochemistry, and pharmacological properties of Echinophora species. The information of this review was obtained by searching for keywords Apiaceae, Echinophora, pharmacological effects, and traditional and modern medicine in scientific articles and books published in search engines Scopus, Google Scholar, Science Direct, PubMed, and Web of Science. The traditional uses of Echinophora and the existence of valuable phytochemicals in the plant have led to isolation and drug discovery of natural medicines such as antibiotic, analgesics, and anticancer drugs, and the beneficial effects of these plants can widely be used in healthcare.
  2,256 40 -
ORIGINAL ARTICLES
Effect of the administration of Solanum nigrum fruit on prevention of diabetic nephropathy in streptozotocin-induced diabetic rats
Fariba Azarkish, Kobra Hashemi, Ardashir Talebi, Mohammad Kamalinejad, Nepton Soltani, Nima Pouladian
October-December 2017, 9(4):325-332
DOI:10.4103/pr.pr_47_17  PMID:29263625
Background: Our previous study showed antidiabetic effect of aqueous extract of Solanum nigrum Linn fruit (SNE). Objective: This study was designed to explore the antidiabetic and nephroprotective effects of SNE in diabetic rats. Materials and Methods: Animals were divided into nine groups to undergo two experiment protocols: Two groups served as nondiabetic controls (NDCs), while the other groups had diabetes induced with a single injection of streptozotocin. SNE-treated diabetic (D-SNE) and SNE-treated controls (NDC-SNE) received 1 g/L of SNE added to the drinking water and insulin-treated diabetic (D-I) for 8 weeks. Furthermore, there were four groups (D-SNE, NDC-SNE, D-I, D) in the second protocol to examine diabetic nephropathy (DN) for 16 weeks. Blood urea nitrogen (BUN), creatinine (Cr) magnesium, nitric oxide (NO), and malondialdehyde (MDA) levels were measured. Both kidneys were isolated to measure MDA, NO levels, and renal damage. Results: SNE could decrease blood glucose level in diabetic rats. In addition, SNE was more effective than insulin in controlling blood glucose. SNE could decrease BUN, Cr levels, and kidney weight and damage after 8 and 16 weeks of administration. Plasma and kidney levels of NO and MDA also decreased. Conclusion: Our results support the hypothesis that SNE could play a role in the management of diabetes and the prevention of DN. Abbreviations Used: SNE: Extract of Solanum nigrum Linn fruit, NDCs: Nondiabetic controls, STZ: Streptozotocin, D-SNE: SNE-treated diabetic, NDC-SNE: SNE-treated controls, D-I: Insulin-treated diabetic, BUN: Blood urea nitrogen, Cr: Creatinine, Mg: Magnesium, NO: Nitric oxide, MDA: Malondialdehyde, DN: Diabetic nephropathy, BW: Body weight, FBG: Fed blood glucose, KW: Kidney weight, TBA: Thiobarbituric acid, IPGTT: Intraperitoneal glucose tolerance test, AUC: Aria under the curve, GFR: Glomerular filtration rate.
  2,126 24 -
Annonacin exerts antitumor activity through induction of apoptosis and extracellular signal-regulated kinase inhibition
Chee Voon Yap, Kavita S Subramaniam, Sik Wey Khor, Ivy Chung
October-December 2017, 9(4):378-383
DOI:10.4103/pr.pr_19_17  PMID:29263632
Background: Endometrial cancer (EC) is the most common gynecologic malignancy in developed countries. Annonacin, a natural pure compound extracted from the seeds of Annona muricata, is a potential alternative therapeutic agent to treat EC. Objective: To study the antitumor activity of annonacin and its mechanism of action in EC cells (ECCs). Materials and Methods: Viability of ECCs treated with annonacin for 72 h was determined using methyl thiazolyl tetrazolium assay. The induction of cell cycle arrest and apoptotic cell death was evaluated using propidium iodide and annexin V-PE/7-AAD assay, respectively. DNA strand breaks were visualized using transferase dUTP nick end labeling assay, and the effects of annonacin on survival signaling were determined using western blotting. Results: Annonacin exhibited antiproliferative effects on EC cell lines (ECC-1 and HEC-1A) and primary cells (EC6-ept and EC14-ept) with EC50values ranging from 4.62 to 4.92 μg/ml. EC cells were shown arrested at G2/M phase after treated with 4 μg/ml of annonacin for 72 h. This led to a significant increase in apoptotic cell death (65.7%) in these cells when compared to vehicle-treated cells (P < 0.005). We further showed that annonacin-mediated apoptotic cell death was associated with an increase in caspase-3 cleavage and DNA fragmentation. Cell apoptosis was accompanied with downregulation of extracellular signal-regulated kinase survival protein expression and induction of G2/M cell cycle arrest. Conclusion: Annonacin may be a potential novel therapeutic agent for EC patients. Abbreviations Used: 7-AAD: 7-Amino-Actinomycin, ATP: Adenosine diphosphate, BSA: Bovine serum albumin, DNA: Deoxyribonucleic acid, EC: Endometrial cancer, ECC-1: Endometrial cancer cell-1, EC50: Half maximal effective concentration, Ept: Epithelial, FBS: Fetal bovine serum, HEC-1A: Human endometrial carcinoma-1A, MTT: Methyl thiazolyl tetrazolium, NaCl: Sodium chloride, NADH: Nicotinamide adenine dinucleotide, RPMI 1640: Roswell Park Memorial Institute Medium, SDS: Sodium dodecyl sulfate
  1,898 28 -
Antioxidant activity and phenolic and flavonoid contents of the extract and subfractions of Euphorbia splendida Mobayen
Zahra Kefayati, Saeed Mohammadi Motamed, Asie Shojaii, Mitra Noori, Roshanak Ghods
October-December 2017, 9(4):362-365
DOI:10.4103/pr.pr_12_17  PMID:29263629
Introduction: The harmful action of the free radicals which cause the oxidative stress can be blocked by antioxidant substances, and different plant extracts showed antioxidant activity. The aim of this study is was evaluation the antioxidant activity of total methanol extract (ME) and subfractions of Euphorbia splendida Mobayen. Materials and Methods: Aerial part of E. splendida was extracted by maceration with methanol and then subfractionated by liquid–liquid fractionation using petroleum ether, chloroform, ethyl acetate, and water. Antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assay, reduction of ferric ions and ferrous ion chelating potential. Total phenolic contents (TPC) and total flavonoid contents (TFC) were estimated with Folin-Ciocaltue and aluminum chloride methods, respectively. Results: The findings revealed that E. splendida ME and subfractions showed a dose-dependent antioxidant activity. ME showed the highest antioxidant activity based on total reduction capability and ferrous ions chelating assay tests. Aqueous fraction and then ethyl acetate fraction showed the best IC50in DPPH radical scavenging test in comparison to butylated hydroxytoluene. ME showed the highest value of TPC and TFC (270.74 ± 0.005 mg/g and 208.23 ± 0.007 mg/g, respectively). Conclusion: This study showed that the extract and subfractions of E. splendida have antioxidant activity. The antioxidant activity of the extract and fractions might be attributed to the presence of phenolic compounds. More studies are needed to determine the active antioxidant compounds of this plant. Abbreviations Used: TPC: Total phenolic content, TFC: Total flavonoid content, DPPH: 2, 2'- diphenyl-1-picrylhydrazyl, BHT: Butylated hydroxytoluene, EDTA: Ethylene Diamine Tetra Acetic acid, ME: Total methanol extract, EAF: Ethyl acetate fraction, AQF: Aqueous fraction, PEF: Pertolium ether fraction, CHF: Chloroformic fraction
  1,871 29 -
Toxicological evaluation of the aqueous extract of Caralluma europaea and its immunomodulatory and inflammatory activities
Zineb Issiki, Chaimaa Moundir, Farida Marnissi, Nadia Seddik, Naima Benjelloun, Younes Zaid, Mounia Oudghiri
October-December 2017, 9(4):390-395
DOI:10.4103/pr.pr_24_17  PMID:29263634
Background: Caralluma europaea (CE) has been studied for its chemical constituents, and no information is available on its toxicity or its pharmacological activities. Objective: To determine the toxicity of an aqueous extract of CE stems in vitro and in vivo after acute and subchronic oral gavages in Swiss albino's mice and its immunomodulatory and inflammatory activities. Materials and Methods: The extract was administrated in single oral dose at 5 g/kg body weight for the acute toxicity test and by gavages daily at doses of 1, 2.5, or 5 g/kg for 30 consecutive days for the subchronic toxicity test. The immunomodulatory activities and inflammatory activities were tested by the evaluation of hemagglutination antibodies (HAs) titers and delayed-type hypersensitivity (DTH) response. Results: For the dose of 1 g/kg, no visible toxic effects were observed. However, for the higher doses, clinical observations of toxicity were noted after 1 week of treatment. This was confirmed by the biochemical parameters values and the histology analyses of the spleen, liver, and kidney tissues. The high cellular mortality rate in vitro when treated with CE extract confirmed their toxicity potential. There was also increase of “HA titer” and “DTH” response in mice treated with nontoxic dose of CE (1 g/kg) compared to control group. This immune activity was confirmed by the high number of lymphocytes infiltrates noted in the different organs. Conclusion: We conclude that CE at the dose up of 1 g/kg produced toxic effect in mice that induced an immune inflammatory reaction. Abbreviations Used: CE: Caralluma europaea, ALT: Alanine aminotransferase, AST: Aspartate aminotransferase, RRBCs: Rat red blood cells, DTH: Delayed-type hypersensitivity response, PBS: Phosphate buffer solution.
  1,849 29 -
Antioxidant and intracellular reactive oxygen species/reactive nitrogen species scavenging activities of three porcupine bezoars from Hystrix brachyura
Peng Nian Yew, Wai Leng Lee, Yau Yan Lim
October-December 2017, 9(4):366-371
DOI:10.4103/pr.pr_145_16  PMID:29263630
Background: Porcupine dates are phytobezoar stones that are used in Traditional Chinese Medicine (TCM) treatments against cancer, postsurgical recovery, dengue fever, etc. The medicinal values have not been scientifically investigated due to the availability and high pricing of the dates. Objectives: This paper represents the first report on the phytochemical content, in vitro antioxidant and intracellular reactive oxygen species (ROS)/reactive nitrogen species (RNS) scavenging properties of the extracts of three porcupine dates: grassy date (GD), black date (BD), and powdery date (PD). Materials and Methods: Dried samples were extracted with methanol and lyophilized. Samples were screened for phytochemical constituents, in vitro antioxidant assays based on total phenolic content (TPC), free radical scavenging, and ferric reducing power (FRP) as well as intracellular ROS and RNS scavenging properties. Results: Phytochemical screening and total tannins assay revealed that tannins, cardiac glycosides, and terpenoids were found in all porcupine dates with tannins forming the major portion of the TPC. In comparison to GD, BD and PD were found to contain significantly high TPC, radical scavenging activity, and FRP. At 200 μg/ml, BD and PD remarkably scavenged 2, 2-azobis (2-amidinopropane) dihydrochloride-induced ROS in RAW264.7 cells and significantly reduced nitric oxide in lipopolysaccharide-stimulated cells. Conclusion: Overall, BD and PD exhibited promising in vitro antioxidant as well as intracellular ROS/RNS scavenging properties. Abbreviations Used: TCM: Traditional Chinese Medicine, BD: Black date, GD: Grassy date, PD: Powdery date, TPC: Total phenolic content, FRS: Free radical scavenging, FRP: Ferric reducing power, NO: Nitric oxide, ROS: Reactive oxygen species, RNS: Reactive nitrogen species, GAE: Gallic acid equivalent, AAE: Ascorbic acid equivalent, PVPP: Polyvinylpolypyrrolidone, DCFH-DA: Dichloro-dihydro-fluorescein diacetate, AAPH: 2, 2-azobis (2-amidinopropane) dihydrochloride, LPS: Lipopolysaccharide
  1,821 33 -
Teucrium polium-induced vasorelaxation mediated by endothelium-dependent and endothelium-independent mechanisms in isolated rat thoracic aorta
Saeed Niazmand, Elahe Fereidouni, Maryam Mahmoudabady, Mahmoud Hosseini
October-December 2017, 9(4):372-377
DOI:10.4103/pr.pr_140_16  PMID:29263631
Objective: There are some reports on hypotensive and antispasmodic effects of Teucrium polium L. (Lamiaceae) (TP). Subjects and Methods: The activity of different concentrations of TP extract (1, 2, 4 and 8 mg/ml) was evaluated on contractile responses of isolated aorta to potassium chloride (KCl) and phenylephrine (PE). Results: The cumulative concentrations of the extract induced a concentration-dependent relaxation in the aorta precontracted by PE and KCl. Extract-induced vasorelaxations in denuded aortic rings precontracted by PE and KCl at lower concentrations were considerably less than intact aortic rings, but this effect was significantly more at concentrations of 4 mg/ml for PE-, 4 and 8 mg/ml for KCl-induced contractions. All the extract concentrations (except 1 mg/ml) significantly relaxed PE-induced contraction in the presence of NG-nitro-L-arginine methyl ester. Indomethacin reduced effectively extract-induced vasorelaxation at 1 and 2 mg/ml. The extract reduced PE- and KCl-induced contractions in the presence of cumulative calcium concentrations and after incubation with diltiazem; this vasorelaxant effect of TP was decreased. TP-induced relaxation was inhibited by heparin, ruthenium red, glibenclamide, and tetraethylammonium, but 4-aminopyridine had no effect on TP-induced relaxation. Conclusion: TP extract has vasorelaxant effect on isolated rat thoracic aorta which mediated by endothelium-dependent and endothelium-independent mechanisms. The relaxation mainly was mediated by inhibition of calcium influx in vascular smooth muscle cells. It seems that the vasorelaxant effect of extract at lower concentrations was mediated by nitric oxide and prostacyclin. Abbreviations Used: ROCC: Receptor operated calcium channels, VDCC: Voltage dependent calcium channels, PLC: Phospholipase C, IP3: 1,4,5 triphosphate inositol, IP3R: IP3 receptors, SR: sarcoplasmic reticulum, RYR: ryanodine receptors, K+ATP: ATP-sensitive potassium channel, K+Ca: Calcium-activated potassium channel, cAMP: Cyclic adenosine monophosphate, cGMP: Cyclic guanosine monophosphate, PGI2: Prostaglandin I2, NO: Nitric oxide
  1,735 26 -
Bioassay-guided In vitro study of the antimicrobial and cytotoxic properties of the leaves from Excoecaria lucida Sw
Ania Ochoa-Pacheco, Julio César Escalona Arranz, Munyewu Beaven, Renato Peres-Roses, Yordania Matos Gámez, Miladis I Camacho-Pozo, Gabriel Llauradó Maury, Maíra Bidart de Macedo, Paul Cos, Josean Fechine Tavares, Marcelo Sobral Da Silva
October-December 2017, 9(4):396-400
DOI:10.4103/pr.pr_124_16  PMID:29263635
Background: Excoecaria lucida Sw. (Euphorbiaceae) is a plant conventionally used throughout the Caribbean in the treatment of infectious diseases. Objective: To evaluate, using bioassay-guided fractionation, the in vitro cytotoxicity and antimicrobial activity of E. lucida leaves. Materials and Methods: A 95% ethanol crude extract was dried and fractionated by solid-liquid separation in four phases (hexane, dichloromethane, ethyl acetate, and butanol). Antimicrobial activity (3 bacteria, 6 yeasts, and 2 fungi) was evaluated by the dilution method with resazurin (2048, 512, 128, 32, and 8 μg/mL). The cytotoxicity assays were evaluated in two cell lines: MRC-5 and RAW 264.7; calculating the selectivity index. Assays were performed for the total extract, the isolated compound with the highest yield, and the ethyl acetate and butanol phases. Isolated compounds were characterized by nuclear magnetic resonance and mass spectrometry techniques. Results: Fractionation process led to the isolation of ellagic acid (784.29 mg), 3,3',4'-tri-O-methyl ellagic 4-O-β-D-glucopyranoside acid (6.1 mg), and corilagin (6.91 mg). The most active were ethyl acetate phase and ellagic acid with IC50= 128 μg/mL against seven and five different species of microorganisms, respectively. The total extract (IC50=512 μg/mL) and the ethyl acetate phase (IC50=128 μg/mL) were cytotoxic in both cell lines, while butanol phase and ellagic acid both with IC50>2048 μg/mL seemed to be safer. Conclusions: The results obtained indicate that the Excoecaria leaves can be conventionally used as antimicrobial, but it should be present that some cytotoxicity could appear. In addition, the three identified compounds were reported for the first time in the species. Abbreviations Used: MRC-5-SV2: Diploid human lung fibroblasts cells, RAW 264.7: Murine macrophages cells, IC50: Inhibitory Concentration 50%, ATCC: American Type Culture Collection, CCEBI: Culture Collection of Industrial Biotechnology Center, CECT: Spanish Culture Collection Type, CFU: Colony forming units, CC50: 50% cytotoxic concentration, CO2: Carbon dioxide, SI: Selectivity index, IR: Infrared spectroscopy, 1H NMR: Nuclear Magnetic Resonance of hydrogen, 13C NMR: Nuclear Magnetic Resonance of carbon, HMQC: Heteronuclear Multiple-Quantum Correlation, HMBC: Heteronuclear Multiple Bond Correlation, COSY: Correlation Spectroscopy, NOESY: Nuclear Overhauser Effect Spectroscopy, KBr: Potassium bromide, DMSO-D6: Deuterated dimethyl sulfoxide, LC-MS: Liquid Chromatography-Mass Spectrometry, [α]D: Optical rotation, EL1: ellagic acid, EL2: 3,3',4'-tri-O-methyl ellagic 4-O-β-D-glucopyranoside acid, EL3: corilagin, Active (+), inactive (−).
  1,698 29 -
Inhibitory effects of Pterodon emarginatus bean oil and extract on Staphylococcus aureus
VS Mendes, JB Sant'Anna, S. C. C. Oliveira, Iriani Rodrigues Maldonade, Eleuza Rodrigues Machado
October-December 2017, 9(4):348-353
DOI:10.4103/pr.pr_13_17  PMID:29263627
Background: Pterodon emarginatus is a tree of the Brazilian Savannah. The beans of this tree are used in folk medicine as anti-inflammatory preparations, especially for infections caused by Staphylococcus aureus. These bacteria can cause simple infections or serious illnesses such as pneumonia, meningitis, endocarditis, toxic shock syndrome, septicemia, and others. Objective: This study had the goal of verifying the effect of the essential oil (OE) from P. emarginatus on the inhibition of S. aureus in culture medium, i.e., “ in vitro” tests. Materials and Methods: The vegetable material was cut and crushed with a press. The OE was obtained by extraction using hexane, alcohol, and water. The P. emarginatus extracts obtained were used to evaluate the antimicrobial effect on S. aureus (ATCC 25923) by tests of well diffusion, disc diffusion, and microdilution. The strain used in the assays was maintained in brain heart infusion broth and nutrient agar until testing. Afterward, the bacteria were spread on agar plates with Mueller-Hinton agar medium. In the wells and on the paper discs, the OE suspensions were placed in the following volumes: 10, 15, 20, 25, 30, 40, and 80 μL and subsequently they were incubated at 35°C ± 2°C. After 24 h, the number of colony-forming unit was determined. Results: Pure OE and hydroalcoholic extract inhibited the growth of S. aureus, while aqueous extract had no effect on bacterial growth in all microbial methods used. Conclusion: Thus, the present study showed the potential of sucupira-based extracts against S. aureus growth, opening new perspectives for the evaluation of these bioactive compounds as phytopharmaceutical products. Abbreviations Used: OE: Essencial oil; AC: Hydroalcoholic oil extract; AQ: Aqueous extracts; MIC: Minimum inhibitory concentration; MBC: Minimum bactericidal concentration; CFU: Colony formed unit.
  1,685 25 -
Identification and characterization of Memecylon species using isozyme profiling
TR Bharathi, Shailasree Sekhar, N Geetha, SR Niranjana, HS Prakash
October-December 2017, 9(4):408-413
DOI:10.4103/0974-8490.218499  PMID:29263637
Background: The protein/isozyme fingerprint is useful in differentiating the species and acts as a biochemical marker for identification and systematic studies of medicinal plant species. Objective: In the present study, protein and isozyme profiles for peroxidase, esterase, acid phosphatase, polyphenol oxidase, alcohol dehydrogenase, and alkaline phosphatase of five species of Memecylon (Melastomataceae), Memecylon umbellatum, Memecylon edule, Memecylon talbotianum, Memecylon malabaricum, and Memecylon wightii were investigated. Materials and Methods: Fresh leaves were used to prepare crude enzyme extract for analyzing the five enzymes isozyme variations. Separation of isozymes was carried out using polyacrylamide gel electrophoresis (PAGE) and the banding patterns of protein were scored. Pair-wise comparisons of genotypes, based on the presence or absence of unique and shared polymorphic products, were used to regenerate similarity coefficients. The similarity coefficients were then used to construct dendrograms, using the unweighted pair group method with arithmetic averages. Results: A total of 50 bands with various Rf values and molecular weight were obtained through PAGE analysis. Among the five Memecylon species, more number of bands was produced in M. wightii and less number of bands was observed in M. edule. The results of similarity indices grouped M. malabaricum and M. wightii in one cluster with 98% similarity and M. umbellatum, M. edule, and M. talbotianum are grouped in another cluster with 79% similarity showing close genetic similarities which is in accordance with the morphological identification of Memecylon species. Conclusion: The protein/isozyme fingerprint is useful in differentiating the species and acts as a biochemical marker for identification of Memecylon species. Abbreviations Used: SDS-PAGE: Sodium docecyl sulfate polyacrylamide gel electrophoresis; NTSYS PC2: Numerical taxonomy system, version 2.2 for Windows XP, Vista, Win7, Win 8 and Win10 including 64 bit
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Comparative physicochemical evaluation of kharekhasak (Tribulus terrestris linn.) before and after mudabbar process
Abdullah Tauheed, Hamiduddin , Salma Khanam, Mohd Akhtar Ali, Mohammad Zaigham
October-December 2017, 9(4):384-389
DOI:10.4103/pr.pr_16_17  PMID:29263633
Background and Objectives: Mudabbar/Tadbeere advia is referred to the processes performed on the drugs to detoxify, purify, and enhance therapeutic action and to reduce its doses before making the formulations in Unani medicine. It improves quality of drugs either by optimizing its desirable characteristics or minimizing the undesirable ones; it makes drug effective, safe, and specific. There is a need of comparative evaluation to understand its significance. Tadbeer of Kharekhasak (KK) khurd (Tribulus terrestris Linn. fruit) is described by Rabban Al-Tabari in Firdausul Hikmat, Akbar Arzani in Qarabadeene Qadri, etc., during the compounding of aphrodisiac formulations. Mudabbar Kharekhasak (MKK) used in Safoofe Kharekhasak mentioned in Al-Qarabadeene was evaluated in this work. Methods: Mudabbar/Tadbeer process was carried out by blending fresh KK. Juice with powdered dry KK and drying it under the sun. Juice used for process is thrice the weight of dry KK powder. The KK before and after the process was evaluated using physicochemical tests: powder characterization, extractive value, alcohol and water soluble matter, ash value, loss on drying (LOD) at 105°C, pH, high-performance thin layer chromatography (HPTLC) fingerprinting, and diosgenin content. Results: Powder characterizations were set in. Increase in successive and nonsuccessive extractive values in various solvents, water/alcohol-soluble content, total ash, acid-insoluble ash, water-soluble ash, and sulfated ash of MKK was noted in comparison with KK. Decrease in LOD at 105°C and pH of MKK powder was observed. HPTLC fingerprinting data were developed for the identification and evaluation. Quantification of diosgenin content increased to 432.1 g/g in MKK as compared to 144.5 g/g in KK, suggesting significant increase in saponin content. Conclusion: Data obtained clearly indicated changes in MKK validating the classical Mudabbar process, probably to enhance/modify the action of drug. Standards for crude and MKK were established for future reference. Abbreviations Used: KK: Kharekhasak, TT: Tribulus terrestris, MKK: mudabbar Kharekhasak, SK: Safoofe Kharekhasak, LOD: loss of weight on drying, HPTLC: High performance thin layer chromatography, BSS: British standard sieve, μl: microliter, SEM: Standard error of mean, nm: nanometer, g: gram.
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