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Origin Al Article International Journal of Medicine and Pharmaceutical Sciences (IJMPS) ISSN (P): 2250-0049; ISSN (E): 2321-0095 Vol. 10, Issue 3, Jun 2020, 35-44 © TJPRC Pvt. Ltd. INVIVO GLYCEMIC AND TOXICITY EFFECTS OF AQUEOUS EXTRACTS OF URTICA MASSAICA NJOROGE GK1, NJAGI ENM2, GIKONYO NK3 & PIERO MN4 1Critical Care Unit, Kenyatta National Teaching, Research and Referral Hospital, Nairobi, Kenya 2,4Department of Biochemistry, Microbiology and Biotechnology, Kenyatta University, Nairobi, Kenya 3Department of Pharmacognosy and Pharmaceutical Chemistry, Kenyatta University, Nairobi, Kenya ABSTRACT Phytotherapy as traditional, alternative or complementary medicine is the most popular therapeutic intervention for noncommunicable diseases.Urtica massaica has been used for various ailments; one among them is diabetes mellitus. However, concerns over the blood glucose lowering potential as well as its safety for human use has not been scientifically demonstrated. The objective of this study was to evaluate the invivo glycemic and toxicity effects of aqueous extracts of Urtica massaica. In evaluation of efficacy, aqueous extract ofUrtica massaica at dose levels of 50, 90.9, 165.1 and 300mg/kg body weight was orally administered into Alloxan induced diabetic rats with a fasting blood glucose equal to or above 200 mg/L. Glycemia was determined hourly and the results obtained after a 24 hour experimental period were Original Article compared to normal (non diabetic) and glibenclamide treated rats. Safety aspects were evaluated by orally administering either dose of 300, 448.14, 669.4 or 1000mg/kg body weight of the plant’saqueous extracts for 28 days. Mean blood glucose values and results on the effect on body weight, blood, liver, pancrease, heart and renal system relative to the control. We determined presence of minerals, phytonutrients and phytochemicals in the plants extracts, which was established using standard methods. Urtica massaica aqueous extracts have considerable antihyperglycaemic activity and is a promising antidiabetic monotherapy. Flavonoids, alkaloids, saponins, steroids, cardiac glycosides, minerals and amino acids were present in the aqueous extracts. The current study validates the use of Urtica massaicaas as it has been utilized for mitigation of diabetes and its continued use is recommended. However, its ethno botanical use can be enhanced by bioassay guided fractionation of bioactive compounds and translational clinical research is recommended. KEYWORDS: Invivo, Urtica Massaica, Glycaemia, Ethnobotanical & Bioactive Compounds Received: Mar 28, 2020; Accepted: Apr 18, 2020; Published: May 29, 2020; Paper Id.: IJMPSJUN20205 INTRODUCTION Urtica massaica is a species of flowering rhizomatous perennial herb plant that grows up to 2 meters tall. It belongs to the kingdom plantae, family Urticaceae (nettle) and genus Urtica. It is commonly known as stinging nettle or forest nettle and largely spread across East Africa. The plant has heart-shaped leaves covered with stinging hairs and serrated edges. This plant is used for food and medicine in several African nations (PROTA, 2004; Alphonse, 2008). In Kenya, it is used as a tonic, to treat stomach ache, diabetes or fractures and venereal diseases.The current study evaluates the invivo effectiveness of Urtica massaica as an antihyperglycaemic agent and safety concerns related to its chronic consumption. METHOD Plant Material www.tjprc.org [email protected] 36 Njoroge GK, Njagi ENM, Gikonyo NK & Piero MN Leaves of Urtica Massaica were harvested in a farm at Arkroad Village, Aboni sublocation in Nyeri County of Kenyaat the foot of Aberdare forest O020’25”S, 36o50’55”E. They were dried under a shade and grounded into a powder using a disc mill. The powder was then packed into airtight light proof containers and stored in a cool place until use. The powder was placed in water and left in a water bath at 60oC for 12 hours, decanted, filtered and lyophylized. Lyophylized material was kept in a refrigerator at 4oC until use. Effect on Glycaemia Swiss albino rats were bred in the Animal house at the Department of Biochemistry, Microbiology and Biotechnology of Kenyatta University, Nairobi Kenya. Eight week old rats weighing 90-140g and a mean weight of 120 grams were used for the study. Hyperglycemia was induced by a single intraperitoneal administration of 186.9 mg/kg body weight (bwt) of freshly prepared 10% alloxan monohydrate (2,4,5,6 tetraoxypyrimidine; 5-6-dioxyuracil) (Etuk 2010, Njoroge et. al., 2017) into rats pre fasted for 12 hours. The animals were given feeds and water ad libitum. Development of hyperglycemia was evaluated 72 hours post alloxan administration and after overnight fasting using a glucometer. Rats with a fasting blood glucose level above 200 mg/L (11.1mmol/L) were considered diabetic and used in this study (Soltesova, 2011; Karau et. al., 2012, Njoroge et. al., 2017). Glibenclamide; an oral antidiabetic drug was used at 3mg per kilogram bwt as the standard drug. Five experimental rats were placed in seven treatment groups. Group I (normal control) consisted of normal rats. Diabetic rats were randomly placed into group II to VII. Group II and III were negative and positive controls respectively. Physiological saline (0.5 ml) was administered to rats in Groups I and II. Group III was administered with the drug; Glibenclamide. Groups IV to VII were administered with therapeutic doses of aqueous extracts of Urtica massaica at 50, 90.9, 165.1 or 300 mg/kg bwtre constituted in 0.5 ml physiological saline. The experimental animals were denied feeds but allowed free access to water until the end of the 24 hours experiment. Blood glucose determination using precise glucose testing meter from Roche® diagnostic was done at 0, 1, 2, 3, 4,6, 12 and 24 hours by nipping the tail of the rats. Toxicity Evaluation Five eight week old normal Swiss albino rats were randomly placed into five groups for logarithmic dose toxicity evaluation. Respective treatments were administered for 28 days. Group I was administered with 0.5 ml physiological saline. Groups II, III, IV and V were administered with the reconstituted aqueous extract at dose levels of 300, 448.14, 669.4 and 1000 mg/kg bwt respectively. During the experimental period, the rats were allowed free access to mice pellet and water and observed for any manifestation of illness, change in behaviour and mortality. The body weight of each rat was determined prior to commencement of extract administration and administered weekly during the dosing period. On the 28th, the rats were euthanized. Blood samples for hematology and biochemistry assays were taken.The liver, kidney, heart, spleen, brain and lungs were collected for necropsy. Individual organs were weighed, the tissues gross anatomy observed for any histopathological changes and stored in 10% formalin. Pathological studies done on blood samples were total blood count (TBC) for hematological studies. Alanine aminotransferase (AST), aspartate aminotransferase (ALT), alkaline phosphatase (ALP), γ-Glutamyltransferase (GGT), total protein and albumin were done in an evaluation of liver function. Blood urea nitrogen (BUN), creatinine (CREAT), sodium (Na+) and potassium (K+) were done in an evaluation of effects on the kidneys, amylase (AMY) and lipase in evaluation of the pancrease and creatinine kinase (CK) as well as lactate dehydrogenase (LDH) in evaluation of effects on Impact Factor (JCC): 7.8387 NAAS Rating: 4.14 Invivo Glycemic and Toxicity Effects of Aqueous Extracts of Urtica Massaica 37 the cardiac system. RESULTS In Vivo Hypoglycemic Effects of Aqueous Extracts of U. Massaica in Alloxan Induced Diabetic Rat Models The four therapeutic doses of U. massaica when orally administered caused a hypoglycemic effect in the alloxan induced diabetic rat model which was not significantly different (p > 0.05) to that of glibenclamide throughout the experimental period (Table 1). The blood glucose lowering effect of the plant extract was not dose dependant (p > 0.05). Further, the blood glucose lowering rate by the plant extract was not significantly different (p > 0.05) from the standard oral antidiabetic drug (Figure 1). An hour post treatments administration, the extract reduced glycemia to 87.6%, 89.3%, 83.1% and 87% at dose levels of 50, 90.9, 165.1 and 300 mg/kg bwt respectively while glibenclamide reduced the blood glucose to 87.3% in the same hour. Table 1: Effects of Orally Administered Aqueous Extract of U. Massaica at Therapeutic doses on Blood Glucose Levels in alloxan Induced Diabetic Rats Treatme Blood Glucose Levels at Varying Times (mmol/l) nt Group 0hr 1hr 2hr 3hr 4hr 6hr 12hr 24hr Normal 5.8±0.4aB 5.8±0.2aB 5.3±0.7abB 5.4±0.7abB 5.2±0.9abC 4.6±0.5abB 4.5±0.5bB 4.3±0.8bB control Negative 23.6±3.4a 23.7±3.6a 22.3±4.99a 16.1±3.7ab 12.6±2.4 11.1±1.9 22.5±4.5aA 20.9±4.8aA control A A A A bA bA Positve 27.5±4.1a 23.7±2.3a 19.9±3.8bc 17.1±2.7bc 13.8±4.5cd 9.2±5.6deA 5.0±1.8eB 4.8±1.5eB control A bA A A AB B Therapeutic aqueous extract doses (mg/kg bwt) of U. massaica 22.5±1.4a 19.8±2.6a 16.1±3.6bc 12.4±3.3cA 50 17.8±2.6bA 5.5±1.2dB 4.1±0.5dB 5.0±0.9dB A bA A BC 26.1±4.6a 23.3±3.8a 22.1±4.3ab 19.98±3.5a 16.6±3.6bc 11.1±4.9cd 90.9 5.3±1.6dB 6.4±4.4dB A bA A bA AB AB 25.7±5.6a 21.0±4.7a 19.98±4.5a 18.1±5.7ab 14.2±5.8bc 165.1 8.2±3.3cdB 4.2±2.1dB 4.4±2.6dB A bA bA A AB 22.3±2.96 19.5±4.1a 17.98±4.4a 15.8±5.3ab 10.7±5.1bcB 300 5.7±5.0cB 4.0±2.8cB 3.5±2.0cB aA A bA A C Values are expressed as Mean ±Standard Deviation (SD) blood glucose of five rats per group.
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