Asian Journal of Research and Reports in Endocrinology

Protective Effect of Methanolic Extract of Vernonia amygdalina (Bitter-Leaf) on Alloxan-Induced Pancreatic Toxicity in Adult Wistar Rats

Christian Chiemeka Ozor

Department of Anatomy, Faculty of Basic Medical Sciences, College of Medicine, Enugu State University of Science and Technology (ESUT), Enugu State, Nigeria.

Frank Onyedikachi Anioba

Department of Anatomy, Faculty of Basic Medical Sciences, College of Medicine, Enugu State University of Science and Technology (ESUT), Enugu State, Nigeria.

Chiadikobi Lawrence Ozoemena *

Department of Anatomy, Faculty of Basic Medical Sciences, College of Medicine, Enugu State University of Science and Technology (ESUT), Enugu State, Nigeria.

*Author to whom correspondence should be addressed.

---

Abstract

The pancreas is a glandular organ endowed with two main functions; an exocrine function that assists digestion and an endocrine function that modulates blood glucose concentration. Pancreatic Beta-cells produce insulin to regulate/lower blood glucose concentration, driving them into cells to be utilized. Distortions of the architecture of pancreatic Beta-cells lead to function loss; thus, preservation/restoration of their cyto-architecture would sustain their functions. Vernonia amygdalina is a vital plant acknowledged widely for its antioxidant and anti-diabetic effects; however, its impact on the pancreas has received little attention. This study is premeditated to examine the effects of the methanolic leaf extracts of Vernonia amygdalina (MLEVA) on the cyto-architecture of the pancreas in Alloxan-induced pancreatic toxicity. Thirty (30) adult male Wistar rats were grouped into 6 (A-F) (n=5). Group A, the control group, received feed and water only. Group B received a single intra-peritoneal injection of 150mg/kg of Alloxan. Groups C, D, and E received single intra-peritoneal injections of 150mg/kg of Alloxan and were then treated with 200, 300, and 400 mg/kg/day of the MLEVA, respectively. Group F received a single intra-peritoneal injection of 150mg/kg of Alloxan and was orally treated with 100mg/kg/day of Vitamin E. The experiment lasted 16 days before sacrificing the animals via a median incision on the abdominal cavity, under ketamine (100mg/ml), as anaesthesia, 24 hours after their last treatment. The pancreas was carefully and rapidly processed for routine H & E staining. This study recorded a 28% mortality rate after Alloxan administration, leading to degenerative histo-pathological changes in the pancreas' endocrine and exocrine compartments. 300mg/kg of MLEVA demonstrated a marked regenerative effect on the pancreas. The MLEVA is a promising agent for managing Diabetes, evidenced by its protective and therapeutic effect on Alloxan-induced pancreatic toxicity.

Keywords: Vernonia amygdalina, MLEVA, pancreatic toxicity, alloxan, diabetes mellitus

---

---

---

References

Vasudeva N, Mishra S. Inderbir Singh's Textbook of Human Histology (Seventh edition): 2014; 298-302.

Marron P, Leslie R, Henri-Jean G, Chaim J, Manuel R, Maria T, Martinez L, Wei-Ping T, Yongsoo P, Zhi-Xing Z. Insulin-dependent diabetes mellitus (IDDM) is associated with CTLA4 polymorphisms in multiple ethnic groups. Human Molecular Genetics. 1997; 6(8):1275–1282.

Wiberg E. Pancreatic acinar atrophy in German shepherd dogs and rough‐coated Collies. Etiopathogenesis, diagnosis and treatment. A review, Veterinary Quarterly. 2004; 26(2):61-75.

Farombi O, Owoeye O. Antioxidant and chemoprotective properties of Vernonia amygdalina and Garcinia biflavonoid. Int J Environ Res Public Health. 2011;8(6): 2533-2555

Ezuruike U, Prieto J. The use of plants in the traditional management of Diabetes in Nigeria: Pharmacological and toxicological considerations. Journal of Ethnopharmacology. 2014;155:857–924

Biser A. Really wild remedies-medicinal plants used by animals. Zooger. 1998;27(1):80-84.

Akah A, Okafor L. Blood sugar lowering effect of Vernonia amygdalina Del, in an experimental rabbit model. Phytotherapy Research. 1992; 6:171–173.

Nimenbo-Uadia R. Effect of Vernonia amygdalina in alloxan-induced diabetic albino rats. Journal Med Lab Sci. 2003; 12(1):25-31.

Gyang S, Nyam D, Sokomba E. Hypoglycemic activity of Vernonia amygdalina (Chloroform extract) in normoglycaemic and Alloxan-induced hyperglycemic rats. Journal of Pharmacy and Bioresources. 2005;1(1):61-66.

Fatmawaty Y, Nina H, Muhammad H, Vivitri P. Antidiabetic and Antioxidant Potential of Vernonia amygdalina Leaf Extract in Alloxan-Induced Sprague-Dawley Rats. OnLine Journal of Biological Sciences. 2020;20(4):190-200.

Sano T, Kiyokazu O, Tetsuro M, Isao N. Giant Mitochondria in Pancreatic Acinar Cells of Alloxan-Induced Diabetic Rats. Toxicologic Pathology. 2010;38:658-665.

Lenzen S. The mechanisms of Alloxan- and streptozotocin-induced Diabetes, Diabetologia. 2008; 51(2): 216–226.

Szkudelski T. The Mechanism of Alloxan and Streptozotocin Action in B Cells of the Rat Pancreas. Physiol Res. 2001;50(6): 536-46.

Heikkila E, Cabbat S. Protection against Alloxan-induced Diabetes in mice by the hydroxyl radical scavenger dimethylurea. Eur J Pharmacol.1978;52:57–60.

Fisher J. Drugs and chemicals that produce Diabetes. Trends Pharmacol Sci. 1985;6:72-5.

Jorns A, Tiedge M, Lenzen S, Munday R. Effect of superoxide dismutase, catalase, chelating agents, and free radical scavengers on the toxicity of Alloxan to isolated pancreatic islets in vitro. Free Radic Biol Med. 1999;26:1300–4.

Mir S, Darzi M. Histopathological abnormalities of prolonged Alloxan-induced diabetes mellitus in rabbits. International Journal of Experimental Pathology. 2009;90:66–73.

Scarpelli G. Toxicology of the pancreas. Toxicol Appl Pharmacol. 1989;101:543–54.

Cheekati R, Rao S, Vijayaraghavan R. A histological study of Alloxan-induced Diabetes on experimental male Wistar rats. Natl J Physiol Pharm Pharmacol. 2017; 7(12):1329-1334.

Alvarez C, Lopez A. Effect of Alloxan diabetes on exocrine pancreatic secretion in the anaesthetized rabbit. Int J Pancreatol. 1989;5:229–38.

Tiscornia M, Janowitz D, Dreiling A. The effect of Alloxan upon canine exocrine pancreatic secretion. Am J Gastroenterol. 1968;49:328–40.

Atangwho IE, Ebong PE, Egbung E, Obi AU. Extract of Vernonia amygdalina Del.(African Bitter Leaf) Can Reverse Pancreatic Cellular Lesion after Alloxan Damage in the Rat. Australian J of Basic and Appl Sci. 2010; 4(5):711-716.

Oyedeji O, Bolarinwa F, Akintola M. Effect of methanol extract of Vernonia amygdalina on haematological and plasma biochemical parameters in male albino rats. Int. J. Dent. Med. Sci. 2013;3(5): 64-67.

Adefisayo M, Rufus O, Akomolafe S, Akinsomisoye K, Alabi L, Ogundipe G, Kehinde P. Protective Effects of Methanolic Extract of Vernonia amygdalina (del.) Leaf on Aspirin-Induced Gastric Ulceration and Oxidative Mucosal Damage in a Rat Model of Gastric Injury. Dose-Response: An International Journal. 2018; 3:1-11.

Bacevic M, Eric R, Regis R, Pierre D, France L. Practical considerations for reducing mortality rates in Alloxan-induced diabetic rabbits. Heliyon. 2020; 6(e04103):1-7.

Akhtar S, Khan A, Malik T. Hypoglycaemic activity of Alpinia galangal rhizome and its extracts in rabbits, Fitoterapia. 2002;73(7–8):623–628.

Díez R, García J, Diez J, Sierra M, Sahagún M, Calle P. The hypoglycemic and hypolipidemic potential of a high fibre diet in healthy versus diabetic rabbits, Bio. Med. Res. Int; 2013;1–8.

Alam S, Khan H, Sirhindi G, Khan S. Alloxan-induced Diabetes in rabbits. Pakistan J. Pharmacol. 2005;22(2): 41–45.

O'Loughlin A, Kulkarni M, Vaughan E, Creane M, Liew A, Dockery P. Autologous circulating angiogenic cells treated with osteopontin and delivered via a collagen scaffold enhance wound healing in the Alloxan-induced diabetic rabbit ear ulcer model. Stem Cell Res. Ther. 2013;4(6): 158.

El-Esawy H, Alghamdy N, El-Askary A, Elsayed M. Histopathological Evaluation Of The Pancreas Following Administration Of Paricalcitol In Alloxan- Induced Diabetic Wistar Rats. World Journal of Pharmacy and Pharmaceutical Sciences. 2016; 5(3):1-10.

Takemoto K, Wakana D, Noriyoshi M. Protective effect of vitamin E against Alloxan-induced mouse hyperglycemia. Biochimica et Biophysica Acta. 2016;1862:647–650.

Malek M, Ali Noorafshan H, Hamid R, Mohammad A, Monireh M, Morteza A, Maryam H. Effects of vitamin E on ovarian tissue of rats following treatment with p-nonylphenol: A stereological study. Iranian Journal of Reproductive Medicine. 2010; 8(1):1-9.

Nwanjo U. Efficacy of aqueous leaf extract of Vernonia amygdalina on plasma lipoprotein and oxidative status in diabetic rat models. Niger J Physiol Sci. 2005; 20(1-2):39-42.

Abraham O. Effects of Vernonia amygdalina and chlorpropamide on blood glucose. Med J Islamic World Acad Sci. 2007;16:115-119.

Ebong E, Atangwho J, Eyong U, Egbung E. The anti-diabetic efficacy of combined extracts from two continental plants: Vernonia amygdalina (A. Juss) and Vernonia amygdalina (del.) (African bitter leaf). Am J Biochemist Biotechnol. 2008; 4(3):239-244.

Uchenna O, Chinwe O, John O, Ijeoma E. Hypoglycemic indices of Vernonia amygdalina on postprandial blood glucose concentration of healthy humans. Afr J Biotechnol. 2008;7:4581-4585.

Erasto P, Grierson D, Afolayan J. Antioxidant constituents in Vernonia amygdalina leaves. Pharm Biol. 2007; 45(3):195-199.

Oboh G, Raddatz H, Helen T. Antioxidant properties of polar and non-polar extracts of some tropical green leafy vegetables. J Sci Food Agric. 2008; 88(14): 2486-2492.

Fasakin F, Aluko C. Antioxidant properties of chlorophyll enriched and chlorophyll depleted polyphenolic fractions from leaves of Vernonia amygdalina and Gongronema latifolium. Food Res Int. 2011; 44(8):2435-2441.