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 MASSAICA

NJOROGE GK1, NJAGI ENM2, GIKONYO NK3 & PIERO MN4 1Critical Care Unit, Kenyatta National Teaching, Research and Referral Hospital, Nairobi, 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 Original 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 ’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 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 (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

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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. Means that share a superscripted capital letter in a column or a superscripted small letter in a row are not significantly different (p≤ 0.05 by ANOVA and Tukey’s post hoc test).

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Figure 1: Change in Blood Glucose Levels by Orally Administered Aqueous Extracts of U. massaica at therapeutic doses in Alloxan Induced Diabetic Rats. NC = Normal control, DNC = Diabetic negative control, DPC = Diabetic positive control (glibenclamide treated), UM = U. massaica. 50, 90.9, 165.1 and 300 represents doses administered at milligram per kilogram bwt respectively.

Evaluation of uptake Safety of Aqueous Extracts of U. Massaica in swiss Albino Rats Effects of Orally Administered Aqueous Extract of U. Massaica on the General bwt of Swiss Albino Rats

Orally administered extract of U. massaica at dose levels of 300, 448.14, 669.4 and 1000 mg/kg bwt did not affect the general body weight of the rats differently (p > 0.05) from the control (Table 2).

Table 2: Effects of Orally Administered High Non-Therapeutic doses of Aqueous Extract of U. massaica on the Body Weight of Rats Treatment Weekly Weights of Rats (g) Average 0 (Baseline) 1 (Day 7) 2 (Day 14) 3 (Day 21) 4 (Day 28) Weekly Weight Change (g) Control 108.5±8.5aB 138.0±18.2aAB 151.0±23.8aAB 161.5±31.5aA 167.3±29.9aA 11.8±11.2a Extract dose (mg/kg body weight) 300 107.6±7.9aC 130.6±4.4aB 143.2±7.9aB 163.4±10.5aAB 169.0±11.1aA 12.8±9.3a 448.14 106.2±12.3aB 123.0±19.4aAB 137.0±28.8aAB 159.4±32.5aA 160.2±33.5aA 10.8±9.97a 669.4 104.0±10.6aC 127.8±11.2aBC 140.6±9.9aAB 170.2±17.9aA 170.8±30.1aA 13.4±13.4a 1000 112.3±8.3aC 135.8±7.1aBC 149.5±10.2aAB 165.0±12.3aAB 167.3±24.6aA 10.95±9.8a

Results are expressed as Mean ± Standard Deviation (SD) for five rats per group. Values that share a superscripted letter are not significantly different at p ≤ 0.05 by ANOVA and Tukey post hoc test. Small letter superscripts in a column indicate the relationship between groups. Capital letter superscripts in a row indicate the relationship in change of individual group’s weight during the period of the experiment.

Effects of Orally Administered Aqueous Extract of U. Massaica on the Organs and Percentage Organ Weights of Swiss Albino Rats

Orally administered U. massaica at doses of 300, 448.14, 669.4 and 1000 mg/kg bwt did not affect the weight of the heart,

Impact Factor (JCC): 7.8387 NAAS Rating: 4.14 Invivo Glycemic and Toxicity Effects of Aqueous Extracts of Urtica Massaica 39 kidney, lungs, spleen and brain relative to the control. However, statistical computation of the percentage organ to bwt indicated that administration of 300mg/kg bwtU. massaica reduced the weight of the liver and spleen significantly compared to the control. There was no effect on the percentage organ to bwt that resulted from oral administration of U. massaica at 448.14, 669.4 and 1000mg/kg bwt compared to the control (Table 3).

Table 3: Effects of Orally Administered High Non-Therapeutic doses of Aqueous Extract of U. Massaica on the Organ and Percentage Organ to Body Weights of Rats Treatment Organ Weight (g/100g) Heart Kidney Lungs Spleen Liver Brain Control 0.6±0.1a 1.5±0.3a 1.6±0.5a 1.2±0.3a 9.1±1.4a 1.2±0.2a (0.4±0.0)a (0.9±0.1)a (0.9±0.2)a (0.7±0.2)a (5.5±0.5)a (0.8±0.2)a Extract dose (mg/kg body weight) 300 0.6±0.1a 1.4±0.1a 1.7±0.1a 0.7±0.1a 6.1±0.9b 1.3±0.2a (0.3±0.0)a (0.8±0.0)a (1.0±0.1)a (0.4±0.1)b (3.6±0.4)c (0.8±0.1)a 448.14 0.6±0.1a 1.3±0.2a 1.6±0.6a 1.1±0.4a 7.3±1.6ab 1.3±0.2a (0.4±0.1)a (0.8±0.1)a (1.0±0.1)a (0.7±0.2)a (4.6±0.6)ab (0.9±0.3)a 669.4 0.6±0.1a 1.4±0.3a 1.7±0.1a 0.9±0.2a 7.4±1.6ab 1.5±0.1a (0.4±0.1)a (0.8±0.0)a (1.0±0.2)a (0.5±0.1)ab (4.3±0.4)bc (0.9±0.1)a 1000 0.8±0.2a 1.6±0.3a 2.2±0.9a 0.9±0.2a 6.7±1.3ab 1.6±0.1a (0.5±0.1)a (1.0±0.3)a (1.3±0.6)a (0.5±0.1)ab (4.0±0.6)bc (1.0±0.2)a

Results are expressed as Mean ± Standard Deviation (SD) for five rats per group. In parenthesis are mean percentage organ to body weight. Values that share a superscripted letter down the column are not significantly different at p≤ 0.05 by ANOVA and Tukey’s post ANOVA test.

Effect of Orally Administered Aqueous Extracts of U. Massaica On Hematological Parameters of Swiss Albino Rats

Except the decrease in Mean Corpuscular Volume (MCV) at the dose level of 300 mg/kg body weight, orally administered U. massaicain significantly affected other red blood cells indices and platelets of the treated rats relative to the control (p > 0.05)(Table 4).

Table 4: Effects of Orally Administered High Non-Therapeutic doses of Aqueous Extract of U. Massaica on the Red Blood Cells Indices and Platelets of Rats Red Blood Cells Parameters and Platelets Treatment RBC Hb (g/dL) PCV (%) MCV (fL) MCH (pg) MCHC PLT (×106/L) (g/dL) (×103/L) Control 6.1±1.1a 12.2±0.8a 41.5±9.0a 67.7±3.5a 20.5±4.5a 30.4±7.1a 632±217a (oral) Extracts dose (mg/kg body weight) 300 (oral) 7.3±0.3a 13.5±0.8a 39.98±2.4a 54.6±2.1b 18.5±0.5a 33.9±0.6a 615±240a 448.14 7.3±0.6a 13.3±1.8a 49.1±2.8a 67.2±4.8a 18.4±3.1a 27.1±4.6a 770±244a (oral) 669.4 7.1±0.3a 13.8±0.7a 42.8±4.6a 60.6±5.8ab 19.6±1.6a 32.5±3.3a 759±167a (oral) 1000 (oral) 7.2±0.9a 14.1±1.3a 42.0±4.3a 60.1±3.5ab 19.7±1.8a 33.5±1.3a 647±170a

Results are expressed as Mean ± Standard Deviation (SD) for five rats per group. Values that share a superscripted letter down the column are not significantly different at p≤ 0.05 by ANOVA and Tukey’s post hoc test. RBC = red blood cell count, Hb = hemoglobin, PCV = packed red cell volume, MCV = mean cell volume, MCH = mean corpuscular hemoglobin, MCHC = mean corpuscular hemoglobin concentration, PLT= platelets. www.tjprc.org [email protected] 40 Njoroge GK, Njagi ENM, Gikonyo NK & Piero MN

All four doses of U. massaica orally administered into rats in this study did not have significant difference (p > 0.05) effects on the total white blood cells count (WBC) and differential counts compared to the controls. (Table 5)

Table 5: Effects of Orally Administered High Non-Therapeutic doses of Aqueous Extract of U. Massaica on White Blood Cells and Differential White Cell Count of Rats Treatment WBC Differential leucocytes count (x109/L ) Dose (x109/L) Neutrophils Lymphocytes Monocytes Eosinophils Basophils Control 3.3±1.3a 0.9±0.7a 1.9±1.1a 0.3±0.2a 0.3±0.2a 0.1±0.0a (oral) Extract dose (mg/kg body weight) 300 (oral) 4.3±0.6a 1.5±0.7a 2.5±1.5a 0.2±0.1a 0.4±0.4a 0.0±0.0a 448.14 3.4±0.5a 1.3±0.4a 1.4±0.9a 0.2±0.2a 0.4±0.4a 0.1±0.0a (oral) 669.4 4.2±1.8a 1.2±0.7a 2.5±1.4a 0.2±0.1a 0.3±0.2a 0.0±0.0a (oral) 1000 (oral) 4.1±1.8a 1.1±0.9a 2.5±1.2a 0.3±0.2a 0.2±0.2a 0.1±0.0a

Results are expressed as Mean ± Standard Deviation (SD) for five rats per group. Values that share a superscripted letter down the column are not significantly different at p≤ 0.05 by ANOVA and Tukey’s post hoc test. WBC = white blood cells count.

Effect of Orally Administered High Non-Therapeutic Doses of Aqueous Extracts of U. Massaica on Biochemical Parameters of Swiss Albino Rats

Orally administered U. massaica at doses of 300, 448.14, 669.4 and 1000 mg/kg body weight did not affect lactate dehydrogenase (LDH) and amylase differently (p > 0.05) compared to the control. However, orally administered dose level of 1000 mg/kg body weight of the plant extract increased (p < 0.05) creatine phosphokinase (CK) and decreased (p < 0.05) lipase levels relative to control (Table 6).

Table 6: Effects of orally administered high non-therapeutic doses of aqueous extract of U. massaica on cardiac and pancreatic function tests of rats Treatment Cardiac Pancreatic LDH (IU/L) CK (IU/L) AMYLASE (IU/L) LIPASE (IU/L) Control 1517±814a 909±447b 1346±409a 15.5±1.0a Extract dose (mg/kg body weight) 300 1249±313a 2145±1089ab 1648±804a 13.8±0.8b 448.14 1665±798a 2730±1525ab 1639±321a 13.8±0.5b 669.4 1626±432a 2028±1573ab 1423±229a 14.2±0.8ab 1000 932±305a 3585±350a 1344±366a 13.5±0.6b

Results are expressed as Mean ± Standard Deviation (SD) for five rats per group. Values that share a superscripted letter down the column are not statistically different at p≤ 0.05 by ANOVA and Tukey’s post hoc test. LDH = lactate dehydrogenase and CK = creatine phosphokinase. IU/L = international units per litre.

Table 7 presents the effect on liver function tests upon orally administering the specified four doses of U. massaica. Albumin levels were decreased (p < 0.05) by orally administered dose level of 300mg/kg body weight of the plant extract compared to the control. Other biomarkers of liver function (AST, ALT, ALP, GGT, TP) were insignificantly affected (p > 0.05) relative to the control.

Impact Factor (JCC): 7.8387 NAAS Rating: 4.14 Invivo Glycemic and Toxicity Effects of Aqueous Extracts of Urtica Massaica 41

Table 7: Effects of Orally Administered High Non-Therapeutic doses of Aqueous Extract of U. Massaica on liver Function Tests of Rats Liver Function Test Analytes Treatment AST (IU/L) ALT (IU/L) ALP (IU/L) GGT (IU/L) TP (g/L) ALB (g/L) Control (oral) 415±191a 185±44a 282.5±142a 3.5±2a 69.7±16.3a 34±7.5a Extract dose (mg/kg body weight) 300 (oral) 506±294a 160±71a 153±49a 3.6±2a 65±2.0a 23.2±2.8b 448.14 (oral) 639±271a 180±65a 180±25a 6.2±3a 69.8±3.4a 25.2±3.9ab 669.4 (oral) 717±260a 172±75a 240±63a 6.2±3a 68.2±4.6a 25.4±3.7ab 1000 (oral) 371±91a 141±32a 173±73a 4.0±2a 67.3±2.1a 24.8±2.9ab

Results are expressed as Mean ± Standard Deviation (SD) for five rats per group. Values that share a superscripted letter down the column are not significantly different at p≤ 0.05 by ANOVA and Tukey’s post hoc test. AST = aspartate aminotransferase, ALT = alanine aminotransferase, ALP = alkaline phosphatase, GGT = gamma glutamyl transferase, TP = total protein and ALB = albumin. IU/L= international units per litre.

On evaluation of renal function, the results indicate that orally administered extract of U. massaica at 300, 448.14, 669.5 and 1000 mg/kg body weight doses did not affect urea and sodium levels of the treated rats significantly different (p > 0.05) compared to control rats. The four doses reduced creatinine and potassium levels compared to control (Table 8).

Table 8: Effects of Orally Administered High Non-Therapeutic doses of Aqueous Extract of U. Massaica on Rats Renal Function Tests Treatment Renal Function Test Analytes UREA (mmol/L) Creatinine (µmol/L) Sodium Potassium (mmol/L) (mmol/L) Control (oral) 11.1±2.1a 105.8±18.6a 155.5±1.3a 22.0±7.5a Extract dose (mg/kg body weight) 300 12.0±0.7a 65.4±5.9b 148.1±0.6a 8.1±1.0b 448.14 13.1±0.6a 74.4±11.3b 154.6±7.7a 10.3±2.7b 669.4 10.8±1.4a 64.2±6.9b 150.8±0.9a 11.1±1.9b 1000 11.0±1.7a 66.0±6.2b 150.3±1.5a 10.7±0.6b

Results are expressed as Mean ± Standard Deviation (SD) for five rats per group. Values that share a superscripted letter down the column are not significantly different at p≤ 0.05 by ANOVA and Tukey’s post hoc test.

Table 9 presents the composition of micronutrients in the aqueous extracts of Urtica massaica.

Table 9: Micronutrients Composition of Urtica Massaica Amino Acid Phytochemicals Minerals Asparagine Tannins calcium Threonine Flavanoids copper Glutamine Saponnins iron Alaninie Steroids potassium Valine Phylobatannins Sodium Isoleucine Magnesium Lysine Manganese Argenine Zinc Proline Phosphates Nitrates

DISCUSSIONS

This study was carried out to determine the invivo effects on glycemia by orally administering aqueous extracts of Urtica

www.tjprc.org [email protected] 42 Njoroge GK, Njagi ENM, Gikonyo NK & Piero MN massaica. Toxicity effects that could be attributed to the extracts administration were also evaluated. Results obtained in this study indicate that the aqueous extracts of Urtica massaica has in vivo blood sugar lowering properties. The glycemic decline was not significantly different between doses during the experimental period. Hence, glycemic effect was not dose dependent. Further, effect of orally administered treatments of Urtica massaicaon glycemia was similar to glibenclamide. Glibenclamide is insulinotropic i.e., acts by increasing insulin secretion from pancreatic beta cells which reduces blood glucose. In addition, glibenclamide also acts through reduction of serum glucagon levels in type II diabetes and an increase of insulin action on target tissues. The study results indicate a correlation of antihyperglycaemic activity of glibenclamide and the aqueous extracts of Urtica massaica. Thus, the mode of hypoglycemic action of the plant aqueous extract could be similar to that of glibenclamide.

Preliminary phytochemical screening indicated the presence of phenolic compounds in aqueous extracts of Urtica massaica. The extractshad tannins, flavonoids, saponins, steroids, cardiac glycosides and phylobatannins. The presence of the phytochemicals in plants extracts with antihyperglycemic properties has been documented (Malviyaet. al., 2010; Singh et al., 2012;Karau et al., 2012; Yogeshaet al., 2013; Ayanglaet al., 2016, Njoroge et al., 2017).

Oxidative stress is a consequence of prolonged hyperglycemia. This results to the micro and macrovascular complications experienced in diabetes such as cardiovascular diseases, neuropathy, nephropathy etc (Sarma, 2010; Shradha, 2010). With a diminished endogenous antioxidant systems, exogenous antioxidants plays a big role in scavenging free radicals and neutralizing their cytotoxic effects. Flavonoids are known potent antioxidants. The aqueous extracts of Urtica massaica in this study have been reported to have flavonoids and hence antioxidant activities. Coupled with their hypoglycemic effects, U. massaica has a great advantage while used as hypoglycemic agents than the conventional antidiabetic drugs (Obohet al., 2013; Yogeshaet al., 2014; Chauhan, 2014).

CONCLUSIONS AND RECOMMENDATIONS

This study determined invivo effects to glycemia and safety by orally administering aqueous extracts of Urtica massaica. The results obtained in this study have indicated that the aqueous plant extracts conferred antihyperglycaemic responses in rats. The aqueous plant extract is thus suitable in control of blood glucose in diabetes and the potential free radical scavenging phytochemicals are paramount in preventing diabetic associated complications. Further, the extracts have not exhibited any toxicity effects in all the four high non-therapeutic doses through oral administration. Thus, they are safe for long term human consumption. Nevertheless, it is recommended that the use of Urtica massaica for health benefits be enhanced on a proper regulatory. Post pre-clinical translational clinical studies on Human beings should be conducted to propagate use of Urtica massaicain medical practice. This equally opens the door for pharmaceutical formulations of convenient consumption forms of the plants extracts.

ACKNOWLEDMENTS

The study was funded by the Republic of Kenya Government National Commission for Science Technology and Innovation (NACOSTI/PHD/008).The entire Management and staff of the Kenyatta University Departments of Biochemistry, microbiology and Biotechnology as well as Pharmacognosy and pharmaceutical chemistry tirelessly contributed to the success of this study. Input from the staff of Physiology Department of the University of Nairobi and Kenyatta National Research, Teaching and Referral Hospital, Springs Healthcare are equally appreciated. Logistical and further financial support by Lynda Connections Limited in Nairobi, Kenya is highly appreciated.

Impact Factor (JCC): 7.8387 NAAS Rating: 4.14 Invivo Glycemic and Toxicity Effects of Aqueous Extracts of Urtica Massaica 43

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