Investigating the protective effect of zinc supplementation on streptozotocin-induced pancreatic beta-cell damage via the HMGB1/TLR2/MAPK pathway

Articles in Press

Document Type : Original Article

Authors

1 Faculty of Health Sciences, Department of Nutrition and Dietetics, Demiroglu Bilim University, Istanbul, Türkiye

2 Faculty of Hamidiye Health Sciences, Department of Nutrition and Dietetics, University of Health Sciences, Istanbul, Türkiye

3 Faculty of Medicine, Department of Physiology, Adiyaman University, Adiyaman, Türkiye

4 Health Science Institute, Translational Medicine, Cukurova University, Adana, Türkiye

10.22038/ijbms.2026.92275.19914

Abstract

Objective(s): This study investigated the protective effect of zinc supplementation on insulin release via modulation of the High Mobility Group Box 1 (HMGB1)/Toll-like receptor 2 (TLR2)/mitogen-activated protein kinase (MAPK) signaling pathway in streptozotocin (STZ)-induced pancreatic β-cell injury, a commonly used model of type 1 diabetes.
Materials and Methos: Human pancreatic β-cells (1.1B4) were divided into five groups: Control, Diabetes, Diabetes + Zinc, Diabetes + Box A, and Diabetes + Zinc + Box A. Diabetes was induced with STZ (20 mM, four hours, type-1), followed by treatment with zinc (30 μM) and/or Box A (100 ng/l). Cellular oxidative stress, apoptosis, mitochondrial membrane potential (MMP), cell viability, and insulin secretion were measured. Protein expressions of HMGB1, TLR2/4, extracellular signal-regulated kinases 1/2 (ERK1/2), and c-Jun N-terminal kinase (JNK) were analyzed using western blotting.
Results: Zinc supplementation restored MMP and total antioxidant status (TAS), reduced apoptotic cell death, and enhanced TLR2 and ERK1/2 expression suppressed by STZ. Combined zinc and Box A treatment markedly increased insulin secretion under both hypo- and hyperglycemic conditions.
Conclusion: Zinc exerts protective effects against STZ-induced β-cell damage, likely through modulation of the HMGB1/TLR2/MAPK axis. These findings highlight the therapeutic potential of zinc supplementation in protecting pancreatic β-cell function in diabetes mellitus.

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Main Subjects

References

1. Abel ED, Gloyn AL, Evans-Molina C, Joseph JJ, Misra S, Pajvani UB, et al. Diabetes mellitus—Progress and opportunities in the evolving epidemic. Cell 2024; 187:3789–3820.
2. Lin X, Xu Y, Pan X, Xu J, Ding Y, Sun X, et al. Global, regional, and national burden and trend of diabetes in 195 countries and territories: an analysis from 1990 to 2025. Sci Rep 2020; 10:14790.
3. Cole JB, Florez JC. Genetics of diabetes mellitus and diabetes complications. Nat Rev Nephrol 2020; 16:377–390.
4. Eizirik DL, Pasquali L, Cnop M. Pancreatic β-cells in type 1 and type 2 diabetes mellitus: Different pathways to failure. Nat Rev Endocrinol 2020; 16:349–362.
5. Kim YK, Walters JA, Moss ND, Wells KL, Sheridan R, Miranda JG, et al. Zinc transporter 8 haploinsufficiency protects against beta cell dysfunction in type 1 diabetes by increasing mitochondrial respiration. Mol Metab 2022; 66:101632.
6. Ahmad R, Shaju R, Atfi A, Razzaque MS. Zinc and diabetes: A connection between micronutrient and metabolism. Cells 2024; 13:1359.
7. Chausmer AB. Zinc, insulin and diabetes. J Am Coll Nutr 1998; 17:109–115.
8. Huang Q, Du J, Merriman C, Gong Z. Genetic, functional, and immunological study of ZnT8 in diabetes. Int J Endocrinol 2019; 2019:1524905.
9. Janko C, Filipović M, Munoz LE, Schorn C, Schett G, Ivanović-Burmazović I, et al. Redox modulation of HMGB1-related signaling. Antioxid Redox Signal 2014; 20:1075–1085.
10. Lei XG, Vatamaniuk MZ. Two tales of antioxidant enzymes on β cells and diabetes. Antioxid Redox Signal 2011; 14:489–503.
11. Mousavi SM, Hajishafiee M, Clark CC, do Nascimento IJB, Milajerdi A, Amini MR, et al. Clinical effectiveness of zinc supplementation on the biomarkers of oxidative stress: A systematic review and meta-analysis of randomized controlled trials. Pharmacol Res 2020; 161:105166.
12. Ohly P, Dohle C, Abel J, Seissler J, Gleichmann H. Zinc sulphate induces metallothionein in pancreatic islets of mice and protects against diabetes induced by multiple low doses of streptozotocin. Diabetologia 2000; 43:1020–1030.
13. Kwak MS, Jung SF, Park IH, Shin J-S. The redox-sensitive protein HMGB1: Intracellular and extracellular roles. Exp Mol Med 2026:345–356.
14. Chen X, Ma J, Kwan T, Stribos EG, Messchendorp AL, Loh YW, et al. Blockade of HMGB1 attenuates diabetic nephropathy in mice. Sci Rep 2018; 8:8319.
15. Chung H, Hong SJ, Choi SW, Koo JY, Kim M, Kim H-J, et al. High mobility group box 1 secretion blockade results in the reduction of early pancreatic islet graft loss. Biochem Biophys Res Commun 2019; 514:1081–1086.
16. Matsuoka N, Itoh T, Watarai H, Sekine-Kondo E, Nagata N, Okamoto K, et al. High-mobility group box 1 is involved in the initial events of early loss of transplanted islets in mice. J Clin Invest 2010; 120:735–743.
17. Itoh T, Nitta T, Nishinakamura H, Kojima D, Mera T, Ono J, et al. HMGB1-mediated early loss of transplanted islets is prevented by anti–IL-6R antibody in mice. Pancreas 2015; 44:166–171.
18. Li M, Song L, Gao X, Chang W, Qin X. Toll-like receptor 4 on islet β cells senses expression changes in high-mobility group box 1 and contributes to the initiation of type 1 diabetes. Exp Mol Med2012; 44:260–267.
19. Siddiqui SS, Dhar C, Sundaramurthy V, Sasmal A, Yu H, Bandala-Sanchez E, et al. Sialoglycan recognition is a common connection linking acidosis, zinc, and HMGB1 in sepsis. Proc Natl Acad Sci U S A 2021; 118:e2018090118.
20. Dubey P, Thakur V, Chattopadhyay M. Role of minerals and trace elements in diabetes and insulin resistance. Nutrients 2020; 12:1864.
21. Bjørklund G, Dadar M, Pivina L, Doşa MD, Semenova Y, Aaseth J. The role of zinc and copper in insulin resistance and diabetes mellitus. Curr Med Chem 2020; 27:6643–6657.
22. Jansen J, Karges W, Rink L. Zinc and diabetes—clinical links and molecular mechanisms. J Nutr Biochem 2009; 20:399–417.
23. Feng Y, Qiu W-L, Yu X-X, Zhang Y, He M-Y, Li L-C, et al. Characterizing pancreatic β-cell heterogeneity in the streptozotocin model by single-cell transcriptomic analysis. Mol Metab 2020; 37:100982.
24. Güven C, Taşkın E, Yumrutaş Ö, Şener LT, Dal F, Ahbab M, et al. The investigation of antidiabetic effects of Leontice leontopetalum extract on human pancreatic β cell lines (1.1 B4) treated with streptozotocin. Turk J Agric Food Sci Technol 2018; 6:792–798.
25. Saini KS, Thompson C, Winterford CM, Walker NI, Cameron DP. Streptozotocin at low doses induces apoptosis and at high doses causes necrosis in a murine pancreatic ß cell line, INS‐1. Biochem Mol Biol Int 1996; 39:1229–1236.
26. Nygaard SB, Larsen A, Knuhtsen A, Rungby J, Smidt K. Effects of zinc supplementation and zinc chelation on in vitro β-cell function in INS-1E cells. BMC Res Notes 2014; 7:84.
27. Lin F, Zhang W, Xue D, Zhu T, Li J, Chen E, et al. Signaling pathways involved in the effects of HMGB1 on mesenchymal stem cell migration and osteoblastic differentiation. Int J Mol Med 2016; 37:789–797.
28. Güven C, Dal F, Ahbab MA, Taskin E, Ahbab S, Cinar SA, et al. Low dose monoethyl phthalate (MEP) exposure triggers proliferation by activating PDX-1 at 1.1 B4 human pancreatic beta cells. Food Chem Toxicol 2016; 93:41–50.
29. Taskin E, Guven C, Kaya ST, Sahin L, Kocahan S, Degirmencioglu AZ, et al. The role of toll-like receptors in the protective effect of melatonin against doxorubicin-induced pancreatic beta cell toxicity. Life Sci 2019; 233:116704.
30. Leary N, Pembroke A, Duggan P. Single stable reagent (Arsenazo III) for optically robust measurement of calcium in serum and plasma. Clin Chem 1992; 38:904–908.
31.Janssen J, Helbing A, editors. Arsenazo III: an improvement of the routine calcium determination in serum. Eur J Clin Chem Clin Biochem 1991;29:197-201. 
32. Homsher R, Zak B. Spectrophotometric investigation of sensitive complexing agents for the determination of zinc in serum. Clin Chem 1985; 31:1310–1313.
33. Wang J, Niu Y, Zhang C, Chen Y. A micro-plate colorimetric assay for rapid determination of trace zinc in animal feed, pet food and drinking water by ion masking and statistical partitioning correction. Food Chem 2018; 245:337–345.
34. Alaaeldin R, Bakkar SM, Mohyeldin RH, Ali FE, Abdel-Maqsoud NMR, Fathy M. Azilsartan modulates HMGB1/NF-κB/p38/ERK1/2/JNK and apoptosis pathways during renal ischemia reperfusion injury. Cells 2023; 12:185.
35. Kaur N, Ruiz-Velasco A, Raja R, Howell G, Miller JM, Abouleisa RR, et al. Paracrine signal emanating from stressed cardiomyocytes aggravates inflammatory microenvironment in diabetic cardiomyopathy. Iscience 2022; 25.
36. Prasun P. Role of mitochondria in pathogenesis of type 2 diabetes mellitus. J Diabetes Metab Disord 2020; 19:2017–2022.
37. Nahdi AMA, John A, Raza H. Elucidation of molecular mechanisms of streptozotocin‐induced oxidative stress, apoptosis, and mitochondrial dysfunction in Rin‐5F pancreatic β‐cells. Oxid Med Cell Longev 2017; 2017:7054272.
38. Yang J, Cherian MG. Protective effects of metallothionein on streptozotocin-induced diabetes in rats. Life Sci 1994; 55:43–51.
39. Li J, Chen L, Zhang Y, Zhang WJ, Xu W, Qin Y, et al. TLR4 is required for the obesity-induced pancreatic beta cell dysfunction. Acta Biochim Biophys Sin 2013; 45:1030–1038.
40. Liu Y, Tian Y, Dai X, Liu T, Zhang Y, Wang S, et al. Lycopene ameliorates islet function and down-regulates the TLR4/MyD88/NF-κB pathway in diabetic mice and Min6 cells. Food Funct 2023; 14:5090–5104.
41. Yoo Y-M, Park YC. Streptozotocin-induced autophagy reduces intracellular insulin in insulinoma INS-1E cells. DNA Cell Biol 2018; 37:160–167.
42. Yavuz O, Dincel GC, Yildirim S, El-Ashram S, AlOlayan E. Impact of apoptosis and oxidative stress on pancreatic beta cell pathophysiology in streptozotocin-induced Type 1 diabetes mellitus. Tissue Cell 2024; 91:102552.
43. Jo WS, Kim SD, Jeong SK, Oh SJ, Baik JS, Seo JA, et al. Toll-like receptor 2-mediated ERK activation significantly upregulates interleukin-6 expression in M2-polarized macrophages. Turkish J Biochem 2024; 49:748–756.
44. Sepehri Z, Kiani Z, Nasiri AA, Kohan F. Toll-like receptor 2 and type 2 diabetes. Cell Mol Biol Lett 2016; 21:2.
45. Xiong L, Zhevlakova I, West XZ, Gao D, Murtazina R, Horak A, et al. TLR2 regulates hair follicle cycle and regeneration via BMP signaling. Elife 2024; 12:RP89335.
46. Ferrer R, Soria B, Dawson CM, Atwater I, Rojas E. Effects of Zn2+ on glucose-induced electrical activity and insulin release from mouse pancreatic islets. Am J Physiol 1984; 246:C520–C527.
47. Ghafghazi T, McDaniel ML, Lacy PE. Zinc-induced inhibition of insulin secretion from isolated rat islets of Langerhans. Diabetes 1981; 30:341–345.
48. Jiang Y, Steinle JJ. HMGB1 inhibits insulin signalling through TLR4 and RAGE in human retinal endothelial cells. Growth Factors 2018; 36:164–171.
49. Zhang J, Chen L, Wang F, Zou Y, Li J, Luo J, et al. Extracellular HMGB1 exacerbates autoimmune progression and recurrence of type 1 diabetes by impairing regulatory T cell stability. Diabetologia 2020; 63:987–1001.
50. Zhao Y, Tan Y, Dai J, Li B, Guo L, Cui J, et al. Exacerbation of diabetes-induced testicular apoptosis by zinc deficiency is most likely associated with oxidative stress, p38 MAPK activation, and p53 activation in mice. Toxicol Lett 2011; 200:100–106.
51. Kaneto H, Xu G, Fujii N, Kim S, Bonner-Weir S, Weir GC. Involvement of c-Jun N-terminal kinase in oxidative stress-mediated suppression of insulin gene expression. J Biol Chem 2002; 277:30010–30018.
52. Jiao J, Dou L, Li M, Lu Y, Guo H-B, Man Y, et al. NADPH oxidase 2 plays a critical role in dysfunction and apoptosis of pancreatic β-cells induced by very low-density lipoprotein. Mol Cell Biochem 2012; 370:103–113.
53. Guzmán-Ruiz R, Ortega F, Rodríguez A, Vázquez-Martínez R, Díaz-Ruiz A, Garcia-Navarro S, et al. Alarmin high-mobility group B1 (HMGB1) is regulated in human adipocytes in insulin resistance and influences insulin secretion in β-cells. Int J Obes 2014; 38:1545–1554.
54. Chung H, Hong SJ, Choi SW, Park C-G. The effect of preexisting HMGB1 within fetal bovine serum on murine pancreatic beta cell biology. Islets 2020; 12:1–8.
Iranian Journal of Basic Medical Sciences

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Available Online from 17 May 2026

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