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Original Article PIPER CHABA EXTRACTS with ANTIBIOFILM ACTIVITY INFLUENCE ANTIHYPERGLYCEMIC and ANTIHYPERLIPIDEMIC RESPONSES in DIABETIC WISTER RATS

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Original Article PIPER CHABA EXTRACTS with ANTIBIOFILM ACTIVITY INFLUENCE ANTIHYPERGLYCEMIC and ANTIHYPERLIPIDEMIC RESPONSES in DIABETIC WISTER RATS International Journal of Pharmacy and Pharmaceutical Sciences Print ISSN: 2656-0097 | Online ISSN: 0975-1491 Vol 13, Issue 9, 2021 Original Article PIPER CHABA EXTRACTS WITH ANTIBIOFILM ACTIVITY INFLUENCE ANTIHYPERGLYCEMIC AND ANTIHYPERLIPIDEMIC RESPONSES IN DIABETIC WISTER RATS SULTANA RAJIA1, KHADIZA KHANAM2, UMME FARHANA2, SOHANUR RAHMAN2, RASHIDUL HAQUE1* 1Center for Interdisciplinary Research (CIR), Varendra University, Rajshahi, Bangladesh, 2Department of Pharmacy, Faculty of Science and Engineering, Varendra University, Rajshahi, Bangladesh Email: [email protected] Received: 01 May 2021, Revised and Accepted: 07 Jul 2021 ABSTRACT Objectives: Piper chaba, native to South and Southeast Asia, has been traditionally used as a medicinal plant. Aim of this study was to evaluate the antihyperglycemic and antihyperlipidemic activities of P. chaba root extracts (RE) in streptozotocin (STZ)-induced diabetic rats along with its antimicrobial activity. Methods: Diabetes was induced in Wister rats through the intraperitoneal administration of STZ (50 mg/kg b.w.). Antidiabetic and antilipidemic activities of the RE (in methanol, ethanol, ethyl acetate and distilled water) were evaluated by administering oral dose (200 mg/kg b.w.) for 21 d. Metformin (12.1 mg/kg b.w.) was used as a positive control. Blood samples of rats were drawn by tail vein puncture and cardiac puncture to determine the fasting blood glucose (FBG) and serum level of total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high- density lipoprotein (HDL), respectively. Standard protocols were followed to determine the antimicrobial and antibiofilm activities against two different strains of bacteria. Results: Oral administration of P. chaba RE for 21 d resulted in a significant (p<0.001) decrease in FBG and TC, TG, and LDL levels (p<0.001), when compared to the untreated diabetic rats. Significant (p<0.001) increase of HDL was observed when ethyl acetate and aqueous RE were administered. Out of four, two extracts showed varying antimicrobial activities, particularly against the gram-positive bacteria. Conclusion: It became evident for the first time that P. chaba extracts possess antimicrobial activities and can serve as biochemical compounds with great alternative therapeutic potential in the management of diabetes and hypercholesterolemia. Keywords: Piper chaba, Antidiabetic, Antilipidemic, Antibiofilm © 2021 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijpps.2021v13i9.41950. Journal homepage: https://innovareacademics.in/journals/index.php/ijpps. INTRODUCTION traditional medicines prepared from herbal plants containing phenolic compounds having antidiabetic activity are cost effective In Bangladesh, four hundred and forty nine species of medicinal with fewer side effects [13]. Also, WHO has been encouraging the plants have been enlisted so far [1]. Among them, Piper chaba is a use of traditional medicine to treat life threatening diabetes as well vine in the family of Piperaceae. It is found in the South Western as it’s complications [14]. region of Bangladesh, particularly at Jashore and Khulna regions, and people use it as an additive spice. Extracts of P. chaba possess Plants of the Piperacae family (such as P. betle, P. crocatum, P. phenolic compounds, antioxidants and showed anti-inflammatory, longum, P. nigrum and P. sarmentosum) have been reported for their cytotoxic and anti-bacterial properties [2, 3]. Also, phytochemicals antidiabetic activities. Mixture extracts of P. crocatum leaves and isolated from plants of the Piper species showed anticancer Cinnamomum burmannii bark was reported to enhance the number properties [4, 5]. and reduce the rats’ blood glucose levels [15]. Also, aqueous RE of P. Medicinal plants are of considerable interest to researchers in recent longumof pancreatic was found β cells to in significantly rats, increase decre thease rats’ the blood hyperglycemic insulin levels, and years to treat diabetes mellitus [6]. Diabetes mellitus (DM) is a hyperlipidemic activities in STZ-induced diabetic rats [16]. group of chronic, complex metabolic disorder characterized by Additionally, in another study, aqueous extract of P. nigrum seeds hyperglycemia due to defects in insulin production, insulin and Vinca rosea flowers treatments lead to significant lowering of secretion, and insulin signaling [7]. Insulin is a potent hormone that blood sugar level and reduction in serum lipids [17]. regulates blood glucose level [8]. Globally, diabetes is the leading cause of human death. According to International Diabetes However, the literature review shows that there has been no study Federation (IDF), an estimated 463 million people, as of 2019, had on the effects of P. chaba RE on STZ-induced rats to evaluate their diabetes worldwide (8.8% of the adult population), with type 2 antidiabetic and antihyperlipidemic activities. Hence, this study was diabetes making up about 90% of the cases [9]. According to the undertaken to investigate these activities of P. chaba REs in induced latest World Health Organization (WHO) data, DM-induced death diabetic rats. reached to 28,065 or 3.61% of total deaths in Bangladesh [10]. Diabetes and hyperlipidemia are two factors responsible for the MATERIALS AND METHODS development of cardiovascular diseases. They have severe effect on Collection of samples and preparation of extracts lipid metabolism and show atypical effect on total lipid profile. Retinopathy, cardiovascular diseases, polyurea, polyphasia result Dry roots of P. chaba were purchased from the local market of from long time uncontrolled diabetes [11]. Khulna, Bangladesh. Identification and authentication of the plant material were confirmed at the Department of Botany, University of Besides insulin, the most widely used oral hypoglycemic drugs are Rajshahi, Bangladesh. Insulin sensitizer, insulin secretagogues, a-glucosidase inhibitors, and dipeptidyl peptidase-4 inhibitors. But those drugs have The sample was powdered using mortar and pestle. 250 gm of the remarkable toxicity and side effects [12]. On the contrary, the powder was accurately weighed and separately dissolved with S. Rajia et al. Int J Pharm Pharm Sci, Vol 13, Issue 9, 44-49 methanol (MeOH), ethanol (EtOH), ethyl acetate (EtOAc) and distilled anticoagulant. The serum obtained after centrifugation at 3500 RPM water (Aqueous, AQ), taking 1500 ml solvent and stirred for 72 h. It for 15 min was separated for lipid measurement. The serum was then filtered and the filtrate was concentrated using a rotary concentration of total cholesterol (TC), triglycerides (TG), high evaporator (RE-401, Stuart equipment, Staffordshire, UK) at 30 °C-40 density lipoprotein (HDL)-cholesterol and low density lipoprotein °C. The yield percentages were calculated using the following formula: (LDL)-cholesterol were measured using UV-spectrophotometric Extract yield (%) = R/S × 100 (where R; weight of extracted plant methods. Laboratory kit reagents (Human, Wiesbaden, Germany) residues and S; weight of the plant raw sample) [18]. The yield of following the company’s instruction were used for all biochemical EtOAc, MeOH, EtOH and AQ extracts were 28%, 12%, 20% and 25% analysis and the absorbance was read using a UV-Vis w/w, respectively. The extracts were stored in small bottles at 4 °C. spectrophotometer (Shimadzu UV-1280, Kyoto, Japan). Chemicals/reagents/diagnostic kits Formation of zone of inhibition in disc diffusion assay The antidiabetic drug, metformin was obtained as a gift sample from Evaluation of the antimicrobial activity of all four Piper chaba root Incepta Pharmaceuticals Ltd, Savar, Dhaka, Bangladesh. STZ (Sisco extracts was determined against Bacillus cereus (gram positive) and Research Laboratories Pvt. Ltd. India) and glucose standard strip/kits, Escherichia coli (gram negative) by disc diffusion method [20]. Each and glucometer (GlucoxTD-4183 Blood glucose test strips, Germany). bacterial strain was subcultured overnight at 37 °C in Mueller-Hilton All other chemicals and reagents used were of analytical grade. agar slants. The bacteria were harvested using 5 ml of sterile saline water. Its absorbance was adjusted at 580 nm by a spectrophotometer Experimental animals and ethical permission and diluted to attain a viable cell count of 107 CFU/ml. Filter paper Three-month old healthy male Wister albino rats (180-220 gm body discs were loaded with Piper chaba stem extract at concentrations of weight) collected from the Pharmacology Lab, Department of 200 µg/disc and 400 µg/disc. The plates were kept in the fridge at 4 °C Pharmacy, Jahangirnagar University, Dhaka, Bangladesh. This for 2 h. to permit the diffusion of plant extract. After incubating at 37 investigation was officially recognized by the Ethical Review °C for 24 h, presence of inhibition zones were measured by a Committee, Varendra University, Rajshahi, Bangladesh. Rats were kept transparent scale in mm and recorded as well. in polypropylene cages group wise, 6 animals per cage. Before the Determination of antibiofilm activity experiment, the animals were synchronized to the experimental/ laboratory condition for 7 d. The ambient temperature, humidity and Formation of biofilm biomass was determined through the microtiter 12 h day-night cycles were maintained according to the animal care plate assay in 96-well polystyrene flat bottom plates [20]. After and welfare
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