The anti-diabetic effect of naonoliposmal encapsulated hybrid extract on RIN-5F diabetic cell line

Document Type : Research Paper


Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran



Objective(s): This study investigates the effect of naonoliposmal encapsulated hybrid extract (NEHE) obtained from three plants (green tea, garlic, and chicory) in treating diabetes through expression of PPARgamma, BGK, and GLUT2 genes involved in diabetes and their relationship with miR-27a. 
Materials and Methods: Herbal extracts were encapsulated in liposome. RIN-5F cells were exposed to streptozotocin (STZ) for 24 hr and divided into five groups to induce the diabetes model. Three groups received 0.4, 1, and 2 mg/ml of NEHE, a positive control group received 5 µg/ml of metformin for 72 hr, and a negative control group was only treated with FBS. Then, they were subjected to an MTT assay to check the toxicity of the extract. An immunohistochemical test was performed to check the level of insulin expression in different groups. Real Time-PCR test was performed to check the expression of desired genes. 
Results: MTT assay showed that the NEHE extract had no toxicity on the tested cells. Also, the immunohistochemistry results showed that insulin expression NEHE group was significantly higher than that of the metformin group (P<0.001), indicating the antidiabetic effect of the extract. In all groups treated with the extract, especially with the maximum extract concentration, the expression level of BGK, PPARgamma, miR-27a, and GLUT2 genes was significantly higher than that of the untreated control group. Furthermore, there was no significant difference in the expression of BGK and PPARgamma genes in groups trated with either metformin or plant extract at concentration of 2 mg/ml groups. NEHE showed an improved effect in treating diabetes in a dose-dependent manner by increasing insulin secretion from pancreatic cells. 
Conclusion: As a glucose transporter and sensor, GLUT2 controls the balance between intracellular and extracellular glucose concentrations. When glucose-induced insulin secretion is impaired, garlic can increase the half-life of insulin, while green tea and chicory reduce insulin resistance through the miR-27a pathway.


1. Prevalence and incidence of type 2 diabetes and prediabetes CC Cowie, SS Casagrande, LS Geiss - 2021
2.What is type 2 diabetes? MD Hurtado, A Vella - Medicine, 2019 - Elsevier.
3. Kavishankar G, Lakshmidevi N, Murthy SM, Prakash H, Niranjana S. Diabetes and medicinal plants-A review. Int J Pharm Biomed Sci. 2011;2(3):65-80.
4. Chatterjee S, Khunti K, Davies MJ. Type 2 diabetes. Lancet. 2017;389(10085):2239-2251.
5. Magnuson MA. Glucokinase gene structure: functional implications of molecular genetic studies. Diabetes. 1990;39(5):523-527.
6. Magnuson M, Shelton K. An alternate promoter in the glucokinase gene is active in the pancreatic β cell. J Bio Chem. 1989;4215936:(27)2464.
7. Thorens B. GLUT2, glucose sensing and glucose homeostasis. Diabetologia. 2015;58(2):221-232.
8. Janani C, Kumari BR. PPAR gamma gene–a review. Diabetes  Metab Syndr. 2015;9(1):46-50.
9. Yu Y, Du H, Wei S, Feng L, Li J, Yao F, et al. Adipocyte-derived exosomal MiR-27a induces insulin resistance in skeletal muscle through repression of PPARγ. Theranostics. 2018;8(8):2171.
10. Mukai K, Nagai S, Ohara K. Kinetic study of the quenching reaction of singlet oxygen by tea catechins in ethanol solution. Free Radic Biol Med. 2005;39(6):752-761.
11. Samuel D, Bharali D, Mousa SA. The role of nanotechnology in diabetes treatment: Current and future perspectives. Int J Nanotechnol. 2011;1(8):53-65.
12. Chacko SM, Thambi PT, Kuttan R, Nishigaki I. Beneficial effects of green tea: a literature review. Chin Med. 2010;5(1):1-9.
13. Noroozi, P., Moazenzade, E, Banaei, Mechanism and clinical aspects of the effects of chicory on diabetes. Asian J Res Med Pharm Sci. 2017;1(4):1-11.
14. Hosseini A, Hosseinzadeh H. A review on the effects of Allium sativum (Garlic) in metabolic syndrome. J Endocrinol Invest. 2015;38(11):1147-1157.
15. Noudoost B, Noori N, Amo Abedini G, Gandomi H, Akhondzadeh Basti A, Jebeli Javan A, Ghadami F. Encapsulation of green tea extract in nanoliposomes and evaluation of its antibacterial, antioxidant and prebiotic properties. J Med Plants. 2015 10;14(55):66-78.
16. Fang JY, Hung CF, Hwang TL, Huang YL. Physicochemical characteristics and in vivo deposition of liposome-encapsulated tea catechins by topical and intratumor administrations. J Drug Targeting. 2005 1;13(1):19-27.
17. Bathina, S., N. Srinivas, and U.N. Das, Streptozotocin produces oxidative stress, inflammation and decreases BDNF concentrations to induce apoptosis of RIN5F cells and type 2 diabetes mellitus in Wistar rats. Biochemical and biophysical research communications, 2017; 486(2): 406-413.
18. Gundala NK, Naidu VG, Das UN. Arachidonic acid and lipoxinA4 attenuate streptozotocin-induced cytotoxicity to RIN5 F cells in vitro and type 1 and type 2 diabetes mellitus in vivo. Nutrition. 2017;35:61-80.
19. Jahanfar S, Gahavami M, Khosravi-Darani K, Jahadi M, Mozafari MR. Entrapment of rosemary extract by liposomes formulated by Mozafari method: Physicochemical characterization and optimization. Heliyon. 2021;7(12): 1-9.
20. Patanè G, Piro S, Rabuazzo AM, Anello M, Vigneri R, Purrello F. Metformin restores insulin secretion altered by chronic exposure to free fatty acids or high glucose: A direct metformin effect on pancreatic beta-cells. Diabetes. 2000 1;49(5):735-740.
21. Guillam MT, Hümmler E, Schaerer E, Wu JY, Birnbaum MJ, Beermann F, Schmidt A, Dériaz N, Thorens B. Early diabetes and abnormal postnatal pancreatic islet development in mice lacking Glut-2. Nature genetics. 1997 1;17(3):327-330.
22. Chen T, Zhang Y, Liu Y, Zhu D, Yu J, Li G, et al. MiR-27a promotes insulin resistance and mediates glucose metabolism by targeting PPAR-γ-mediated PI3K/AKT signaling. Aging (Albany NY). 2019;11(18):7510.
23. Damasceno DC, Netto AO, Iessi IL, Gallego FQ, Corvino SB, Dallaqua B, Sinzato YK, Bueno A, Calderon ID, Rudge MV. Streptozotocin-induced diabetes models: pathophysiological mechanisms and fetal outcomes. BioMed research international. 2014;2014:8190.
24. Uldry M, Ibberson M, Hosokawa M, Thorens B. GLUT2 is a high affinity glucosamine transporter. FEBS lett. 2002;524(1-3):199-203.
25.Matschinsky FM. Glucokinase, glucose homeostasis, and diabetes mellitus. Curr Diab Rep. 2005;5A(3):171-176.
26. Sung H-Y, Hong C-G, Suh Y-S, Cho H-C, Park J-H, Bae J-H, et al. Role of (−)-epigallocatechin-3-gallate in cell viability, lipogenesis, and retinol-binding protein 4 expression in adipocytes. Naunyn Schmiedeberg’s Arch Pharmacol. 2010;382(4):303-310. 
27. Hung P-F, Wu B-T, Chen H-C, Chen Y-H, Chen C-L, Wu M-H, et al. Antimitogenic effect of green tea (epigallocatechin gallate) on 3T3-L1 preadipocytes depends on the ERK and Cdk2 pathways. Am J Physiol Cell Physiol. 2005;288(5):C1094-C1108.
28. Wang CT, Chang HH, Hsiao CH, Lee MJ, Ku HC, Hu YJ, et al. The effects of green tea (epigallocatechin‐3‐gallate) on reactive oxygen species in 3T3‐L1 preadipocytes and adipocytes depend on the glutathione and 67 kDa laminin receptor pathways. Mol Nutr Food Res. 2009;53(3):349-360.
29. Eidi A, Eidi M, Esmaeili E. Antidiabetic effect of garlic (Allium sativum L.) in normal and streptozotocin-induced diabetic rats. Phytomed. 2006;13(9-10):624-629.
30. Street RA, Sidana J, Prinsloo G. Cichorium intybus: Traditional uses, phytochemistry, pharmacology, and toxicology. Evid Based Complement Alternat Med. 2013;2013:579319. 
31. Hou X, Tian J, Geng J, Li X, Tang X, Zhang J, et al. MicroRNA-27a promotes renal tubulointerstitial fibrosis via suppressing PPARγ pathway in diabetic nephropathy. Oncotarget. 2016;7(30):47760.
32. Goudappala P, Gowda CY, Kashinath R. Diallyl disulfide regulates purine metabolism and their metabolites in diabetes mellitus. Indian J Physiol Pharmacol. 2021;65(1):28-34.