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Exploration of Antidiabetic Activity of Stephania Japonica Leaf Extract in Alloxan-Induced Swiss Albino Diabetic Mice

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Exploration of Antidiabetic Activity of Stephania Japonica Leaf Extract in Alloxan-Induced Swiss Albino Diabetic Mice Journal of Pharmaceutical Research International 26(6): 1-12, 2019; Article no.JPRI.48311 ISSN: 2456-9119 (Past name: British Journal of Pharmaceutical Research, Past ISSN: 2231-2919, NLM ID: 101631759) Exploration of Antidiabetic Activity of Stephania japonica Leaf Extract in Alloxan-Induced Swiss Albino Diabetic Mice Md. Dobirul Islam1, Syeda Farida Akter1, Md. Amirul Islam1 and Md. Salim Uddin1* 1Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh. Authors’ contributions This work was carried out in collaboration among all authors. Author MDI designed the study, performed the statistical analysis and wrote the first draft of the manuscript. Author SFA managed the literature searches and carried out the tests. Authors MAI and MSU managed the analyses of the study and reviewed the manuscript. All authors read and approved the final manuscript. Article Information DOI: 10.9734/JPRI/2019/v26i630154 Editor(s): (1) Dr. Jongwha Chang, University of Texas, College of Pharmacy, USA. Reviewers: (1) Rajibul Islam, Gono Bishwabidyalay, Bangladesh. (2) Dr. Dennis Amaechi, Veritas University Abuja, Nigeria. Complete Peer review History: http://www.sdiarticle3.com/review-history/48311 Received 24 January 2019 Accepted 07 April 2019 Original Research Article Published 20 April 2019 ABSTRACT Aims: Presently the medicinal world is rapidly turning more on the therapeutic health benefits of natural product and medicinal plants in the management of major crucial disease and their complications. Medicinal plant, Stephania japonica has been studied for exploring antidiabetic potentiality as an alternative source of medicine against the global threat of Diabetes mellitus (DM). Methods: The extraction of S. japonica leaf was carried out by acetone and ethanol. Phytochemical screening and quantitative analysis of S. japonica leaf extracts were evaluated through chemically forming characterized color formation and calibration method respectively, by using standard reference substances (ascorbic acid, gallic acid, and quercetin) to assess the probable compounds present in the extract. Anti-diabetic potentiality of highest phytochemicals containing two extracts were investigated in in vitro as a ⍺-amylase inhibitors and in vivo through alloxan-induced Swiss albino diabetes mice model. _____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected]; [email protected]; Islam et al.; JPRI, 26(6): 1-12, 2019; Article no.JPRI.48311 Results: Alkaloids, carbohydrates, steroids, flavonoids, resins, saponins, tannins and coumarins were present in the leaf extracts. The estimated amount of total phenolics, flavonoids, flavonols and proanthrocyanidins contents of acetone and ethanol extract were 92.12±0.64 and 56.54±1.05 mg of gallic acid equivalent (GAE)/gm of dry extract, 66.02±1.42 and 46.17±0.54 mg of catechin equivalent (CAE)/gm of dry extract, 7.05±0.108 and 5.26±0.083 mg of quercetin equivalent (QUE)/gm of dry extract, 35.19±0.67 and 9.55±1.11 mg CAE/gm of dry extract, respectively. In 3, 5- dinitrosalicylic acid method, acetone and ethanol extract showed α-amylase inhibition of 51.02% and 46.62%, respectively at the concentration of 1000 µg/mL whereas in starch iodine color assay, acetone and ethanol extract showed inhibition of 57.32% and 52.12%, respectively at the concentration of 800 µg/mL. In contrast, both of the leaf extracts significantly (p<0.05 to p<0.001) improved the lipid profile parameters, blood glucose level and serum hepatic marker proteins in alloxan-induced diabetic mice. Conclusion: The present study strongly concluded that S. japonica leaf extracts process potent antidiabetic potentiality that might be significance for the management of diabetes and its complications. Keywords: Diabetic mellitus; phytochemicals; flavonoids; Stephania japonica; hypolipidemic effect. 1. INTRODUCTION consumption on plasma glucose levels [7]. The life threaten effect and detrimental complication Diabetes mellitus, which is reflected as a group of DM are mediated by oxidative stress. In of metabolic disease is considered as one of the diabetic condition, large number of free radicals main leading threat to human lives in the 21st that are produced impaired anti-oxidant defense century. The chronic metabolic disorder, DM is system leading to oxidative damaged through the caused due to deficiency or inadequate function reactive oxygen species mediated diabetic of insulin that leads to abnormalities in glucose pathogenesis [8,9]. Presently many commercial metabolism resulting in ketoacidosis, thirst, synthetic drugs such as biguanides, acarbose, polyuria, blurring of vision, and weight loss [1,2]. glitazones and sulfonylureas are used in the Diabetes also dramatically increases the risk of treatment of DM for reduction of blood glucose various devastating complications such as level. For management of DM and its cardiovascular diseases, peripheral vascular complication many commercial synthetic drugs disease, stroke, neuropathy, renal failure, used throughout the prolong period of life and retinopathy complications and blindness [3,4]. hence produced many adverse side effects [10]. Recently, the number diabetic patients are Therefore, researchers are looking for safe and increasing apprehensively. There are now 415 effective medications to overcome the million adults aged from 20 to 79 years with devastating effects of diabetes. Many medicinal diabetes worldwide and moreover, another 318 plants such as Ficus racemosa, Momordica million adults are estimated to have impaired charantia etc. contain large amount of bioactive glucose tolerance, which puts them at high risk of secondary metabolites that are used in the developing the disease. If this augmentation is treatment of diabetes have high potency of continued for 2040 almost 642 million people will activity, non-toxic, with less or no side effects. be suffering from the diabetes complication [5]. Antioxidant compounds specially, polyphenols in the medicinal plants have been found prolific Inhibitors of amylase based on alkaloids, activity against diabetes [11]. glycosides, galactomannan, terpenoids, peptidoglycans and steroids, are ordinarily Stephania japonica is an evergreen slender prescribed to diabetic patients to decrease twining shrub with greenish yellow flowers and postprandial hyperglycemia induced by the large tubers. The leaf, stem and tuberous root digestion of starch in the small intestine [6]. are astringent, bitter and acrid used in the These inhibitors are designed to primarily target treatment of wide variety of ailments such as of exo-acting starch hydrolyzing enzymes (α- diarrhea, dysentery, fevers, stomach ache, amylase) found in the intestinal tract. The dyspepsia, hepatitis, and urinary diseases inclusive consequence of inhibition is to reduce [12,13]. So far, not enough scientific data has the flow of starch monomers from complex been found regarding antidiabetic properties of dietary carbohydrates into the bloodstream, S. japonica leaf. Therefore, the present study diminishing the postprandial effect of starch was to explore the antidiabetic activity of S. 2 Islam et al.; JPRI, 26(6): 1-12, 2019; Article no.JPRI.48311 japonica leaf extract in alloxan-induced diabetic 2.4 In vitro Antidiabetic (α-amylase mice and the effect on lipid profile and liver inhibition) Assay function biochemical markers were also investigated to ensure the antidiabetic activity of In vitro antidiabetic assay was performed by S. japonica leaf extract. The screening and measuring α-amylase inhibition activity. α- quantification of phytochemicals were performed amylase inhibition assay was carried out using 3, to compare and support these findings. 5-dinitrosalicylic acid (DNSA) method [20] and starch-iodine color assay [21]. 2. MATERIALS AND METHODS 2.4.1 DNSA method 2.1 Collection of Plant Materials and Authentication Ethanol and acetone extracts of S. japonica leaf and standard (acarbose) at the concentration of The fresh leaves of S. japonica were collected 100 µg/mL to 1000 µg/mLwere added to 500 μL from well growth healthy plants from the area of of 0.02 M sodium phosphate buffer (pH-6.9) Rajshahi University campus, Rajshahi (north- containing 6 mM NaCl and 0.5mg/mL of α- western part of Bangladesh) and authenticated amylase solution. Exactly, 500 μL of 1% starch by Prof. Dr. A. H. M. Mahbubur Rahman, solution in 0.02 M sodium phosphate buffer (pH- Taxonomist, Department of Botany, University of 6.9) were added to the mixture and then Rajshahi, Bangladesh. The leaves were washed incubated at 37°C for 10 min. The reaction was with fresh water and then dried under the shade. stopped with 1.0 mL of DNSA reagent and After complete shade drying, the leaves were heated in a boiling water bath for 5 min and then pulverized into a coarse powder with the cooled to room temperature. The reaction help of a grinding machine (FFC-15, China) and mixture was then diluted by the addition of 10 mL were stored in an airtight container for further distilled water and absorbance was measured at use. 540 nm. The control samples were also prepared accordingly without any plant extracts and were 2.2 Preparation of Plant Extracts compared with the test samples containing various concentrations of extracts. The results Approximately, 200 g of powdered leaf material were expressed as % inhibition calculated using was kept immersed in 500 mL of each solvent, the formula: ethanol and acetone
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