Suitability of the Neonatal Streptozotocin Diabetes Model and Folklore Therapeutic use of Low-Dose Neem Leaf Extract to Treat Hyperglycemia Associated with Type 2 Diabetes Mellitus

http://www.doi.org/10.26538/tjnpr/v8i1.30

Authors

  • Garsha A. McCalla Department of Basic Medical Sciences, Faculty of Medical Sciences, University of the West Indies, Mona, Kingston 7, Jamaica.
  • Paul D. Brown Department of Basic Medical Sciences, Faculty of Medical Sciences, University of the West Indies, Mona, Kingston 7, Jamaica.

Keywords:

type 2 diabetes mellitus, hyperglycemia, streptozotocin, neonatal rat model, Neem leaf extract

Abstract

Neem leaf has been used in folklore medicine to combat diabetes mellitus and other ailments. Difficulty finding suitable type 2 diabetes mellitus (T2DM) models led to the exploration of the neonatal streptozotocin (nSTZ) model to investigate the impact of low-dose neem leaf extract (NLE) on T2DM-associated hyperglycemia. Following ethical approval, two- and three-day-old neonatal rat pups (n=66) received 60 mg/kg STZ intraperitoneally while normal controls (n=9) received an equivalent volume of citrate buffer. Weaned animals received free access to chow and water and a constant 12 hour on/12 hour off light cycle. Tail vein blood glucose was assessed weekly following an eight-hour fast, using Accu Chek Advantage glucometer. Animals received tap water (Normal and Diabetic Control) or 0.8 % NLE (Diabetic Treated) for six weeks. The results were expressed as means ± standard error of the mean using the Origin Pro 7.0 software. The significance of the mean difference between groups was determined with the IBM SPSS for Windows software version 23 via the paired samples T-test. Statistical significance of p<0.05 was used. The mortality was 74% pre-STZ (32.6% post-STZ) with 40.9% or 81.8% success (of STZ-injected and STZ-survivors), respectively. Fasting blood glucose was significantly (p<0.05) elevated in the diabetic groups and was not alleviated by low-dose NLE. Because of the variability in mortality and success rates, further investigation is warranted for better reproducibility of the nSTZ T2DM model with minimal mortality. Furthermore, low-dose therapeutic NLE does not positively impact hyperglycemia and should not replace clinically prescribed medicines.

References

American Diabetes Association. Classification and diagnosis of diabetes: standards of medical care in diabetes - 2020. Diabetes Care 2020; 43(Suppl 1): S14-31. https://doi.org/10.2337/dc20-S002

Barragán-Bonilla MI, Mendoza-Bello JM, Aguilera P, Parra-Rojas I, Illades-Aguiar B, Ramírez M, Espinoza-Rojo M. Combined administration of streptozotocin and sucrose accelerates the appearance of type 2 diabetes symptoms in rats. J Diabetes Res 2019; 3: 1-12. https://doi.org/10.1155/2019/3791061

McCalla G. Mortality and success rates of the neonatal rat model of type 2 diabetes mellitus. Endocrinol Diabetes Res 2021; 7: 10. https://www.scitechnol.com/peer-review/mortality-and-success-rates-of-the-neonatal-rat-model-of-type-2-diabetes-mellitus-mfTW.php?article_id=18578

Furman BL. Streptozotocin-induced diabetic models in mice and rats. Curr Protoc 2021; 1: e78. https://doi.org/10.1002/cpz1.78

Anthraa AH. Impact of aqueous extract of neem leaves in lowering blood glucose and lipid profile in STZ induced diabetes mellitus mice. Iraqi J Agric Sci 2022; 53(5): 977-984. https://jcoagri.uobaghdad.edu.iq/index.php/intro/article/view/1611

Rinaldi F, Hanieh PN, Maurizi L, Longhi C, Uccelletti D, Schifano E, Del Favero E, Cantù L, Ricci C, Ammendolia MG, Paolino D, Froiio F, Marianecci C, Carafa M. Neem oil or almond oil nanoemulsions for vitamin E delivery: from structural evaluation to in vivo assessment of antioxidant and anti-inflammatory activity. Int J Nanomedicine 2022; 17: 6447-6465. https://doi.org/10.2147/IJN.S376750

Achi NK, Onyeabo C, Nnate DA, Ekeleme-Egedigwe CA, Kalu IK, Chibundu IC, Wokoma GC. Therapeutic effects of Azadirachta indica A.Juss. leaves in malaria-induced male Wistar rats. J Pharm Pharmacogn Res 2018; 6(3): 191–204. http://jppres.com/jppres/pdf/vol6/jppres17.304_6.3.191.pdf

McCalla G, Parshad O, Brown PD, Gardner MT. Beta cell regenerating potential of Azadirachta indica (Neem) extract in diabetic rats. W Indian Med J 2015; 65(1): 13-17. https://doi.org/10.7727/wimj.2014.224

Parshad O, Singh P, Gardner M, Fletcher C, Rickards E, Choo-Kang E. Effect of aqueous neem (Azadirachta indica) on testosterone and other blood constituents in male rats. A pilot study. West Indian Med J 1994; 43: 71−4.

Bonner-Weir S, Trent DF, Honey RN, Weir GC. Responses of neonatal rat islets to streptozotocin: limited β-cell regeneration and hyperglycemia. Diabetes 1981; 30: 64-9. https://doi.org/10.2337/diab.30.1.64

Kunz L, Richter JS, Mayerhofer A. The adenosine 5'-triphosphate-sensitive potassium channel in endocrine cells of the human ovary: role in membrane potential generation and steroidogenesis. J Clin Endocrinol Metabol 2006; 91(5): 1950-5. https://doi.org/10.1210/jc.2005-2269

Sridevi S, Chary MG, Krishna DR, Diwan PV. Pharmacodynamic evaluation of transdermal drug delivery system of glibenclamide in rats. Indian J Pharmacol 2000; 32: 309-12.

Opazo JC., Soto-Gamboa M, Bozinovic F. Blood glucose concentration in caviomorph rodents. Comp Biochem Physiol A 2004; 137: 57-64. https://doi.org/10.1016/j.cbpb.2003.09.007

Guo XX, Wang Y, Wang K, Ji BP, Zhou F. Stability of a type 2 diabetes rat model induced by high-fat diet feeding with low-dose streptozotocin injection. J Zhejiang Univ Sci B 2018; 19(7): 559-69. https://doi.org/10.1631/jzus.B1700254

Elamin NMH, Fadlalla IMT, Omer SA, Ibrahim HAM. Histopathological alteration in STZ-Nicotinamide diabetic rats, a complication of diabetes or a toxicity of STZ? Int J Diabetes Clin Res 2018; 5(3): 91-98. https://doi.org/10.23937/2377-3634/1410091

Williams M, Asemota HN, Williams E, McGrowder D, Nwokocha C, Voutchkov M, Francis R. The effect of a combination treatment of biomagnetic therapy and a low glycemic influenced diet on nonfasting blood glucose levels in type 2 diabetic rats. Diabetes Manag 2018; 8: 74-81.

Portha B, Picon L, Rosselin G. Chemical diabetes in the adult rat as the spontaneous evolution of neonatal diabetes. Diabetologia 1997; 17: 371-7. https://doi.org/10.1007/BF01236272

Li L, Yi Z, Seno M, Kojima I. Activin A and betacellulin: effect on regeneration of pancreatic beta-cells in neonatal streptozotocin-treated rats. Diabetes 2004; 53(3): 608-15. https://doi.org/10.2337/diabetes.53.3.608

Fang J-Y, Lin C-H, Huang T-H, Chuang S-Y. In vivo rodent models of type 2 diabetes and their usefulness for evaluating flavonoid bioactivity. Nutrients 2019; 11(3): 530-52. https://doi.org/10.3390/nu11030530

Arulmozhi DK, Veeranjaneyulu A, Bodhankar SL. Neonatal streptozotocin-induced rat model of type 2 diabetes mellitus: a glance. Indian J Pharmacol 2004; 36: 217-21. https://www.ijp-online.com/text.asp?2004/36/4/217/11145

Abdel Moaty AA, El-Kholie EA, Adarous RA. The anti-diabetic effect of neem leaves (Azadirachta indica) in alloxan-induced diabetic rats. J Home Econ 2022; 32(2): 19-31

Yarmohammadi F, Mehri S, Najafi N, Salar Amoli S, Hosseinzadeh H. The protective effect of Azadirachta indica (neem) against metabolic syndrome: A review. Iran J Basic Med Sci 2012; 24: 280-92. https://doi.org/10.22038/ijbms.2021.48965.11218

Jayakumar K, Srinivasan M, Ramesh N, Sachan A, Umesh M, Narayana K. Effect of neem leaf extract on feed intake and body weight in rats. Indian Vet J 2002; 79: 732-3.

Pingali U, Ali MA, Gundagani S, Nutalapati C. Evaluation of the effect of an aqueous extract of Azadirachta indica (Neem) leaves and twigs on glycemic control, endothelial dysfunction and systemic inflammation in subjects with type 2 diabetes mellitus - A randomized, double-blind, placebo-controlled clinical study. Diabetes Metab Syndr Obes 2020; 13: 4401-12. https://doi.org/10.2147/DMSO.S274378

El-Hawary ZM, Kholief TS. Biochemical studies on hypoglycemic agents (I) effect of Azadirachta indica leaf extract. Arch Pharm Res 1990; 13: 108–12. https://doi.org/10.1007/BF02857845

Khosla P, Bhanwra S, Singh J, Seth S, Srivastava RK. A study of hypoglycemic effects of Azadirachta indica (Neem) in normal and alloxan diabetic rabbits. Indian J Physiol Pharmacol. 2000; 44(1): 69-74.

Vasudeva Rao A, Ragha Sudha Madhuri V, Rajendra Prasad Y. Evaluation of the in vivo hypoglycemic effect of neem (Azadirachta indica A. Juss) fruit aqueous extract in normoglycemic rabbits. Res J Pharmaceut, Biol Chem Sci 2012; 3(1): 799-806.

Martínez N, Rodríguez Y, Salguero O, Requena D, Triana L, Perez-Ybarra L. A study of hypoglycemic effects of Azadirachta indica (neem) in human blood cells. Emir J Food Agric 2014; 26(7): 623-29. https://doi.org/10.9755/ejfa.v26i7.1819.

Mishra A, Dave N. Neem oil poisoning: case report of an adult with toxic encephalopathy. Indian J Crit Care Med. 2013; 17(5): 321-322. https://doi.org/10.4103/0972-5229.120330

Deng YX, Cao M, Shi DX, Yin ZQ, Jia RY, Xu J, Wang C, Lv C, Liang XX, He CL, Yang ZR, Zhao J. Toxicological evaluation of neem (Azadirachta indica) oil: acute and subacute toxicity. Environ Toxicol Pharmacol. 2013; 35(2): 240-246. https://doi.org/10.1016/j.etap.2012.12.015

Seriana I, Akmal M, Darusman D, Wahyuni S, Khairan K, Sugito S. Neem leaf (Azadirachta indica A. Juss) ethanolic extract on the liver and kidney function of rats. The Sci World J 2021; 2021: 7970424, 7 pages, https://doi.org/10.1155/2021/7970424

Lafta FM, Mohammed RK, Alhammer AH, Ahmed ME. Cytotoxic potential of neem (Azadirachta indica A. Juss) oil. Trop J Nat Prod Res 2023; 7(12): 5436-5440

Trivedi A, Ahmad R, Sahabjada S, Aparna Misra A. Effect of alkaline pH on cytotoxicity profile of neem (Azadirachta indica) ethanolic extract against human breast cancer cell line MDA-MB-231. Eur J Integr Med 2018; 24: 1-7. https://doi.org/10.1016/j.eujim.2018.10.004

Batra N, Kumar VE, Nambiar R, De Souza C, Yuen A, Le U, Verma R, Ghosh PM, Vinall RL. Exploring the therapeutic potential of Neem (Azadirachta Indica) for the treatment of prostate cancer: a literature review. Ann Transl Med 2022; 10(13): 754. https://doi:10.21037/atm-22-94

Nagini S, Palrasu M, Bishayee A. Limonoids from neem (Azadirachta indica A. Juss.) are potential anticancer drug candidates. Med Res Rev 2023; 1-40. https://doi:10.1002/med.21988

Folakemi AV, Abubakar MT, Olawale AS, Dahiru UM, Halima-Sadiya I. Cytogenotoxicity of aqueous Azadirachta indica A. Juss extracts on Nile tilapia – Oreochromis niloticus (Linnaeus, 1758) under static exposure. Trop J Nat Prod Res 2022; 6(7): 1159-1164. https://doi.org/10.26538

Published

2024-02-01

How to Cite

McCalla, G. A., & Brown, P. D. (2024). Suitability of the Neonatal Streptozotocin Diabetes Model and Folklore Therapeutic use of Low-Dose Neem Leaf Extract to Treat Hyperglycemia Associated with Type 2 Diabetes Mellitus: http://www.doi.org/10.26538/tjnpr/v8i1.30. Tropical Journal of Natural Product Research (TJNPR), 8(1), 5925–5931. Retrieved from https://tjnpr.org/index.php/home/article/view/3415