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   Table of Contents - Current issue
July-September 2017
Volume 9 | Issue 3
Page Nos. 221-303

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Uncaria tomentosa (Willd. ex Schult.) DC (Rubiaceae) sensitizes THP-1 cells to radiation-induced cell death p. 221
Lisa Allen, Alison Buckner, Carly A Buckner, Pablo Cano, Robert M Lafrenie
Background: Uncaria tomentosa (Willd. ex Schult.) DC (Rubiaceae), known as Cat's Claw or Uña de gato, is a traditionally used medicinal plant native to Peru. Some studies have shown that U. tomentosa can act as an antiapoptotic agent and enhance DNA repair in chemotherapy-treated cells although others have shown that U. tomentosa enhanced apoptosis. Objective: To determine if treatment with U. tomentosa can significantly enhance cell death in THP-1 cells exposed to ionizing radiation. Materials and Methods: THP-1 monocyte-like cells were treated with ethanolic extracts of U. tomentosa in the presence or absence of bacterial lipopolysaccharide and then exposed to ionizing radiation. Cell proliferation was assessed by MTT and clonogenic assays and the effects on cell cycle measured by flow cytometry and immunoblotting. Changes in cell signaling were determined by immunoblotting and cytokine ELISA and activation of apoptosis measured by caspase activation and DNA fragmentation analysis. Results: Treatment of THP-1 cells with U. tomentosa had a small effect on cell proliferation. However, when the U. tomentosa-pretreated cells were also subjected to 5–9 Gy ionizing radiation, they showed a significant decrease in cell proliferation and increased cellular apoptosis as measured by DNA fragmentation and caspase activation. Treatment with U. tomentosa also decreased the expression of Cyclin E and Cyclin B, key regulators of normal cell cycle progression, and decreased the phosphorylation of various stress-activated, cell survival proteins including p38, ERK, and SAP/JNK kinase. Conclusions: These results suggest that U. tomentosa could be useful in enhancing cell death following anticancer therapies including ionizing radiation.
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High performance thin layer chromatography: Densitometry method for determination of rubraxanthone in the stem bark extract of Garcinia cowa Roxb p. 230
Dachriyanus Hamidi, Hilyatul Aulia, Meri Susanti
Context: Garcinia cowa is a medicinal plant widely grown in Southeast Asia and tropical countries. Various parts of this plant have been used in traditional folk medicine. The bark, latex, and root have been used as an antipyretic agent, while fruit and leaves have been used as an expectorant, for indigestion and improvement of blood circulation. Aims: This study aims to determine the concentration of rubraxanthone found in ethyl acetate extract of the stem bark of G. cowa by the high-performance thin-layer chromatography (HPTLC). Materials and Methods: HPTLC method was performed on precoated silica gel G 60 F254 plates using an HPTLC system with a developed mobile-phase system of chloroform: ethyl acetate: methanol: formic acid (86:6:3:5). A volume of 5 μL of standard and sample solutions was applied to the chromatographic plates. The plates were developed in saturated mode of twin trough chamber at room temperature. The method was validated based on linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and specificity. The spots were observed at ultraviolet 243 nm. Results: The linearity of rubraxanthone was obtained between 52.5 and 157.5 ppm/spot. The LOD and LOQ were found to be 4.03 and 13.42 ppm/spot, respectively. Conclusion: The proposed method showed good linearity, precision, accuracy, and high sensitivity. Therefore, it may be applied for the quantification of rubraxanthone in ethyl acetate extract of the stem bark of G. cowa.
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Investigation of effect of 1,8-cineole on antimicrobial activity of chlorhexidine gluconate p. 234
Merih Simsek, Reşat Duman
Background: Chlorhexidine belongs to a group of medicines called antiseptic antibacterial agents. Chlorhexidine is commonly used for the care and clean off the skin, hands, and wounds. In recent years, medicinal and aromatic plants have been used for prevention of disease, maintaining health, and improving disease in traditional and modern medicine as a medicament. According to recent research, cineole is the isolated active agent of eucalyptus oil and possesses antimicrobial activity. It was demonstrated that cineole could enhance the antimicrobial effects of the other antiseptics. Objective: The aim of this study was to investigate the efficacy of 1,8-cineole on the antimicrobial effect of chlorhexidine against some microorganisms. Materials and Methods: The effect of 1,8-cineole on antimicrobial activity of chlorhexidine gluconate (CHG) was tested using seven different microorganisms. In this study, CHG (128–0.125 mg/l) and cineole (512–2 g/l) were analyzed together and separately using checkerboard assay. Interactions between CHG and 1,8-cineole have been identified as synergistic, indifferent, or antagonistic. Results: Synergistic activity was demonstrated between CHG and 1,8-cineole against Staphylococcus aureus, methicillin-resistant S. aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, and Candida albicans. Indifferent interactions for these compounds were demonstrated against Pseudomonas aeruginosa. Conclusion: CHG antiseptic properties were found to be increased when CHG was used in combination with 1,8-cineole. In this way, CHG will reveal stronger effect against microorganisms.
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Antioxidant and antiproliferative activity of Asparagopsis taxiformis p. 238
PV Neethu, K Suthindhiran, MA Jayasri
Background: Asparagopsis taxiformis (Rhodophyta) is a species of red algae belonging to the family Bonnemaisoniaceae. The objective of the present study was to evaluate antioxidant and antiproliferative activity of four fractions (petroleum ether, chloroform, ethyl acetate, and methanol) of A. taxiformis. Materials and Methods: The red seaweed, A. taxiformis was collected from Mandapam Coastal Region, Gulf of Mannar, Tamil Nadu. Epiphytes present in algal extracts were cleaned and washed with seawater and fresh water. In vitro antioxidant activity was determined by hydrogen peroxide scavenging, ferric reducing antioxidant power, superoxide radical, metal-chelating activity, and phosphomolybdenum reduction assay. Further, the cytotoxic activity was evaluated using brine shrimp lethality assay. This method is rapid, reliable, inexpensive, and convenient as compared to other cytotoxicity assays. Gallic acid, ethylenediaminetetraacetic acid, ascorbic acid, and quercetin were used as reference antioxidant compounds. Results: Reducing power of chloroform extract increased with increasing concentration of the extract. The radical scavenging activity of extracts was in the following order: ascorbic acid > methanol > chloroform > petroleum ether > ethyl acetate. Highest metal-chelating activity was observed in petroleum ether fractions (63%). Reduction of Mo (VI) to Mo (V) increased in methanol extract (27%) at 100 μg/ml. Moreover, all fractions had an inhibitory effect on the formation of hydroxyl radicals. Results showed that ethyl acetate, methanol, and petroleum ether fractions exhibited potent cytotoxic activity with median lethal concentration values of 84.33, 104.4, and 104.4 μg/ml, respectively. Conclusion: Thus, the results showed that red algae possess strong antioxidant and cytotoxic activity that suggests their possible use in the development of pharmaceutical drugs.
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Ameliorative effect of curcumin on olanzapine-induced obesity in Sprague-Dawley rats p. 247
Subramani Parasuraman, Khor Ming Zhen, Urmila Banik, Parayil Varghese Christapher
Objective: To evaluate the effect of curcumin on olanzapine-induced obesity in rats. Materials and Methods: Sprague-Dawley (SD) rats were used for experiments. The animals were divided into six groups, namely, normal control, olanzapine control, betahistine (10 mg/kg), and curcumin 50, 100, and 200 mg/kg treated groups. Except the normal control group, all other animals were administered with olanzapine 4 mg/kg intraperitoneally to induce obesity. The drugs were administered once daily, per oral for 28 days. During the experiment, body weight changes and behavior alterations were monitored at regular intervals. At the end of the experiment, blood sample was collected from all the experimental animals for biochemical analysis. Part of the liver and kidney tissues was harvested from the sacrificed animals and preserved in neutral formalin for histopathological studies. Results: Curcumin showed a significant reduction in olanzapine-induced body weight gain on the rats and improved the locomotor effects. The effect of curcumin on olanzapine-induced body weight gain is not comparable with that of betahistine. Conclusion: This study has shown metabolic alteration effect of curcumin on olanzapine, an antipsychotic drug, treated SD rats.
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Spectrophotometric quantification of flavonoids in herbal material, crude extract, and fractions from leaves of Eugenia uniflora Linn p. 253
Rhayanne T. M. Ramos, Isabelle C. F. Bezerra, Magda R. A. Ferreira, Luiz Alberto Lira Soares
Background: The traditional use of Eugenia uniflora L. (“Pitanga”) is reported due to several properties, which have often been related to its flavonoid content. Objective: The aim was to evaluate analytical procedures for quantification of total flavonoids content (TFCs) by ultraviolet-visible (UV-Vis) spectrophotometry in the herbal material (HM), crude extract (CE), and fractions from leaves of E. uniflora. Materials and Methods: The method for quantification of flavonoids after complexation with aluminum chloride (AlCl3) was evaluated: amount of sample (0.25–1.5 g); solvent (40%–80% ethanol); reaction time and AlCl3concentration (2.5%–7.5%). The procedures by direct dilution (DD) and after acid hydrolysis (AH) were used and validated for HM and CE and applied to the aqueous fraction (AqF), hexane fraction, and ethyl acetate fractions (EAF). Results: The ideal conditions of analysis were ethanol 80% as solvent; 0.5 g of sample; λmax of 408 (DD) and 425 nm (AH); 25 min after addition of AlCl3 5%. The procedures validated for standards and samples showed linearity (R2 > 0.99) with limit of detection and limit of quantification between 0.01 and 0.17 mg/mL (rutin and quercetin); and 0.03 and 0.09 mg/mL (quercetin), for DD and AH, respectively. The procedures were accurate (detect, practice, and repair <5% and recovery >90%), and stable under robustness conditions (luminosity, storage, reagents, and equipment). The TFCs in AqF and EAF were 0.65 g% and 17.72 g%, calculated as rutin. Conclusions: UV-Vis methods for quantification of TFC in HM, CE, and fractions from leaves of E. uniflora were suitably validated. Regarding the analysis of fractions, the EAF achieved enrichment of about nine times in the content of flavonoids.
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In vitro antidiabetic activity of polar and nonpolar solvent extracts from Leucas Aspera (Willd.) link leaves p. 261
VM Annapandian, R Shanmuga Sundaram
Background: Diabetes mellitus is a chronic illness, and the management of diabetes is a global problem. Successful treatment is required to prevent complications and organ damages. Herbal medicines are having minimal adverse effects when compared to the available synthetic drugs to treat such chronic diseases and disorders. Objective: The present study was aimed to evaluate the antidiabetic and antioxidant activity of polar and nonpolar solvent extracts of Leucas aspera (Willd.) link leaves under in vitro models. Materials and Methods: The in vitro antidiabetic activity of petroleum ether (nonpolar) and ethanol (polar) extracts were evaluated in C2C12 cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (cell viability method) and glucose uptake assay. 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging method used for the evaluation of in vitro antioxidant activity. Results: Both the polar and nonpolar solvent extracts of L. aspera had shown better antioxidant activity compared to standard (IC50 = 18.96 and 19.90 μg/mL, respectively). Petroleum ether extract exhibited better cytotoxic activity in C2C12 cell line compared to ethanol extract (concentration of test drug needed to inhibit cell growth by 50% 110.75 ± 5.5 vs. 415.25 ± 8.0 μg/mL) whereas ethanol extract showed enhanced glucose uptake activity than petroleum ether extract in C2C12 cell line at same concentrations. Conclusion: From our study results, we concluded that L. aspera (Willd.) link leaves had shown better antidiabetic activity and antioxidant activity under in vitro models. Nonpolar solvent extract produced slightly better activity than polar solvent extract. This study warrants further research and experiments on animal models.
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Evaluation of antioxidant and hepatoprotective activity of fruit rind extract of Garcinia dulcis (Roxburgh) Kurz p. 266
Nabajyoti Gogoi, Ankur Gogoi, Bijoy Neog, Dibyojyoti Baruah, Khumanthem Deepak Singh
Background: Garcinia spp. belongs to the family Clusiaceae has been traditionally used for the treatment of many ailments including the liver damage. Garcinia dulcis found in North Eastern region of Assam; India can be a potential candidature to combat different ailments. Objective: The present work has been designed in such a way to appraisal the antioxidant and hepatoprotective activity of fruit rind extract of this plant. Materials and Methods: The antioxidant activity was investigated through the various in vitro models, namely, 2,2-diphenyl-1-picrylhydrazine, 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid, nitrite oxide. Phytochemical investigation for total phenolic and flavonoids contents were carried out by standard protocol. For the evaluation of hepatoprotective activity, albino Wistar rats were divided into five groups, five animals per group and activity was determined by measuring the contents of liver function marker enzymes such as serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase (ALP), and biochemical parameter, that is, Bilirubin and total protein. Histopathology observation of liver sections was conducted. Results: Phytochemical investigation revealed the presence of both phenolic and flavonoid groups in the extract in a significant amount. Antioxidant activity of the plant extract was observed in all models and percentage of inhibition was dose-dependent. Intoxicated with carbon tetrachloride, elevated the liver function enzymes, bilirubin, and suppressed the production of total protein. Pretreatment with the extract decreased the SGOT, SGPT, ALP, and bilirubin level significantly and increased the production level of total protein in a dose-dependent manner. The histopathological observation supported the hepatoprotective potentiality of the extract. Conclusion: The results indicate that fruit rind part of G. dulcis is nontoxic and the plant can utilize as an antioxidant source. The plant has a protective agent for liver damages and other diseases caused by free radicals.
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Evaluation of anti-inflammatory and antimicrobial activity of AHPL/AYCAP/0413 capsule p. 273
Sanjay Nipanikar, Sohan Chitlange, Dheeraj Nagore
Background: Conventional therapeutic agents used for treatment of Acne are associated with various adverse effects necessitating development of safe and effective alternative therapeutic agents. In this context, a polyherbal formulation AHPL/AYCAP/0413 was developed for treatment of Acne. Objectives: To evaluate Anti-inflammatory and antimicrobial activity of AHPL/AYCAP/0413. Material and Methods: 1) Anti-inflammatory activity: Anti-inflammatory activity of AHPL/AYCAP/0413 in comparison with Diclofenac was assessed in carrageenan induced rat Paw edema model. 2) Anti-microbial activity for P. acne: Propionibacterium acnes were incubated under anaerobic conditions. Aliquots of molten BHI with glucose agar were used as the agar base. Formulation and clindamycin (10 μg/ml) were introduced in to the Agar wells randomly. 3) Anti-microbial activity for Staphylococcus epidermidis and Staphylococcus aureus: Staphylococcus epidermidis and Staphylococcus aureus were incubated under aerobic conditions at 37°C. TSB with glucose agar was used as the agar base. 0.5ml of formulation and clindamycin (10 μg/ml) were introduced in to the wells randomly. The antibacterial activity was evaluated by measuring zones of inhibition (in mm). Result: Significant reduction in rat paw edema (51% inhibition) was observed with formulation AHPL/AYCAP/0413 which was also comparable to that of Diclofenac (58% inhibition). Zone of inhibition for formulation was 18.33 mm, 19.20 mm and 26.30 mm for P. acnes, S. epidermidis and S. aureus respectively. This activity was also comparable to that of Clindamycin. Conclusion: AHPL/AYCAP/0413 capsule possesses significant Anti-inflammatory and Anti-microbial activities which further justifies its role in the management of Acne vulgaris.
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Development of validated high-performance thin-layer chromatography method for simultaneous determination of quercetin and kaempferol in Thespesia populnea p. 277
Hiteksha Panchal, Aeshna Amin, Mamta Shah
Introduction: Thespesia populnea L. (Family: Malvaceae) is a well-known medicinal plant distributed in tropical regions of the world and cultivated in South Gujarat and indicated to be useful in cutaneous affections, psoriasis, ringworm, and eczema. Bark and fruits are indicated in the diseases of skin, urethritis, and gonorrhea. The juice of fruits is employed in treating certain hepatic diseases. The plant is reported to contain flavonoids, quercetin, kaempferol, gossypetin, Kaempferol-3-monoglucoside, β-sitosterol, kaempferol-7-glucoside, and gossypol. T. populnea is a common component of many herbal and Ayurvedic formulation such as Kamilari and Liv-52. Objective: The present study aimed at developing validated and reliable high-performance thin layer chromatography (HPTLC) method for the analysis of quercetin and kaempferol simultaneously in T. populnea. Method: The method employed thin-layer chromatography aluminum sheets precoated with silica gel as the stationary phase and toluene: ethyl acetate: formic acid (6:4:0.3 v/v/v) as the mobile phase, which gave compact bands of quercetin and kaempferol. Result: Linear regression data for the calibration curves of standard quercetin and kaempferol showed a good linear relationship over a concentration range of 100-600 ng/spot and 500-3000 ng/spot with respect to the area and correlation coefficient (R2) was 0.9955 and 0.9967. The method was evaluated regarding accuracy, precision, selectivity, and robustness. Limits of detection and quantitation were recorded as 32.06 and 85.33 ng/spot and 74.055 and 243.72 ng/spot for quercetin and kaempferol, respectively. Conclusion: We concluded that this method employing HPTLC in the quantitative determination of quercetin and kaempferol is efficient, simple, accurate, and validated.
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The potency of red seaweed (Eucheuma cottonii) extracts as hepatoprotector on lead acetate-induced hepatotoxicity in mice p. 282
Giftania Wardani, Nuraini Farida, Rina Andayani, Mahmiah Kuntoro, Sri Agus Sudjarwo
Background: Lead is one of the most toxic metals, producing severe organ damage in animals and humans. Oxidative stress is reported to play an important role in lead acetate-induced liver injury. Aim: This study was carried out to investigate the role of ethanol extract of Eucheuma cottonii in protecting against lead acetate-induced hepatotoxicity in male mice. Materials and Methods: The sample used fifty male mice which were divided into five groups: negative control (mice were given daily with Aquadest); positive control (mice were given daily with lead acetate 20 mg/kg body weight (BW) orally once in a day for 21 days); and the treatment group (mice were given E. cottonii extracts 200 mg, 400 mg, and 800 mg/kg BW orally once in a day for 25 days, and on the 4th day, were given lead acetate 20 mg/kg BW 1 h after E. cottonii extract administration for 21 days). On day 25, the levels of serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvate transaminase (SGPT), alkaline phosphatase (ALP), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were measured. The data of SGOT, SGPT, ALP, MDA, SOD, and GPx were analyzed with one-way ANOVA, followed by least significant difference test. Results: The results showed that oral administration of lead acetate 20 mg/kg BW for 21 days resulted in a significant increase in SGOT, SGPT, ALP, and MDA levels. Moreover, there was a significant decrease in SOD and GPx levels. Treatment with E. cottonii extracts of 800 mg/kg BW but not with 200 mg/kg BW and 400 mg/kg BW significantly (P < 0.05) decreased the elevated SGPT, SGOT, ALP, and MDA levels as compared to positive control group. Treatment with E. cottonii extracts of 800 mg/kg BW also showed a significant increase in SOD and GPx levels as compared to positive control group. Treating mice with lead acetate showed different histopathological changes such as loss of the normal structure of hepatic cells, blood congestion, and fatty degeneration whereas animals treated with lead acetate and E. cottonii extracts showed an improvement in these changes and the tissue appeared with normal structures. Conclusion: It can be concluded that E. cottonii extracts could be a potent natural product and can provide a promising hepatoprotective effect against lead acetate-induced hepatotoxicity in mice.
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Quantification of quercetin obtained from Allium cepa Lam. leaves and its effects on streptozotocin-induced diabetic neuropathy p. 287
Khan Dureshahwar, Mohammed Mubashir, Hemant Devidas Une
Objective: Antioxidant potential has protective effects in diabetic neuropathy (DN); hence, the present study was designed with an objective to quantify quercetin from shade-dried leaves of Allium cepa Lam. and to study its effects on streptozotocin (STZ)-induced chronic DN. Materials and Methods: The shade-dried leaves of A. cepa Lam. were extracted with methanol and then fractionated using ethyl acetate (ACEA). The quantification of quercetin in ACEA was evaluated by high-performance thin layer chromatography (HPTLC). The STZ (40 mg/kg) was administered to Sprague-Dawley rats (180–250 g) maintained at normal housing conditions. The STZ was administered once a day for 3 consecutive days. The elevation in blood glucose was monitored for 3 weeks periodically using flavin adenine dinucleotide-glucose dehydrogenase method by Contour TS glucometer. Rats showing blood glucose above 250 mg/dl were selected for the study. Animals were divided into eight groups. ACEA (25, 50, and 100 mg/kg), quercetin (40 mg/kg), metformin (120 mg/kg), and gabapentin (100 mg/kg) were given orally once a day for 2 weeks. The blood glucose level was again measured at the end of treatment to assess DN. Thermal hyperalgesia, cold allodynia, motor incoordination, and neurotoxicity were studied initially and at the end of 2-week treatment. Biochemical parameters were also evaluated after 2-week drug treatment. Results: The quercetin present in ACEA was 4.82% by HPTLC. All the ACEA treatment reduces blood glucose level at the end of the 2-week study and shows a significant neuroprotective effect in STZ-induced DN in the above experimental models. Conclusion: The quercetin present in ACEA proved protective effect in STZ-induced DN.
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Root exudates of Cyperus alternifolius in partial hydroponic condition under heavy metal stress p. 294
Boopathy Usharani, Namasivayam Vasudevan
Secondary metabolites play a vital role in the treatment of various ailments as well as in phytoremediation. The link between secondary metabolites and phytoremediation needs exploration. Hitherto, no information is available regarding the phytochemical components that exist in the root exudates of Cyperus alternifolius. This study was designed to determine the phytocomponents in the root exudates of C. alternifolius under heavy metal stress. C. alternifolius was grown by a novel technique in partial hydroponic conditions and imperiled to a mixture of heavy metals (Cd, Cu, Cr, Ni, Zn, Pb, and Fe) at different concentrations. The root exudates were collected, freeze-dried, redissolved and reconstituted in hexane and analyzed in gas chromatography–mass spectrometry using JEOL GCMATE II in SAIF IIT-Madras. The analysis revealed that the profile of phytochemicals in root exudates is diverse with biological properties. Few phytochemicals found in the root exudates are not cited earlier in any literature. The composition and percentage of phytochemicals could not be correlated to heavy metal concentration. Phytochemical composition decreased with an increase in heavy metal concentration. Control plant released more phytochemicals than the plants under heavy metal stress. From the results, it is evident that root exudates of C. alternifolius contain various bioactive components. Further research can be extended to evaluate the pharmaceutical importance of the species and explore its role in phytoremediation of heavy metals.
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Ex vivo antispasmodic activity of aqueous extract of flowers of Muntingia calabura Linn. on excised rabbit's jejunum p. 301
Kannan Vadivel, Gollapudi Sandeep Kumar, Sitty Manohar Babu
Objective: The present study has been undertaken with the main objective of evaluating the aqueous extract of flowers of Muntingia calabura for antispasmodic activity on isolated rabbit's jejunum. Materials and Methods: The study was carried out on isolated rabbit's jejunum preparations. The aqueous extract of flowers of M. calabura was applied in different doses by cumulative manner without washing the tissue. Spontaneous contractions abolished by the extract were recorded. Results: In isolated rabbit's jejunum preparation, the flower extract of M. calabura inhibited the spontaneous contractions in a concentration-dependent manner with IC100value of 36 ± 3.02 μg/mL, which is potent than the standard drug verapamil with IC100value of 40 ± 1.02 μg/mL on the rabbit's jejunum. Conclusion: The aqueous extract of flowers of M. calabura exhibits significant dose-dependent relaxations of spontaneous contractions in isolated rabbit's jejunum preparations.
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