Sex-based differences in streptozotocin-induced type 2 diabetes rat models

Abstract

Background: Diabetic foot ulcers (DFUs) are a severe complication of diabetes mellitus, leading to chronic infections, amputations, and increased mortality despite existing treatments. More effective therapeutics are urgently needed, and animal models provide a critical platform for investigating the molecular mechanisms underlying DFUs. Streptozotocin (STZ)—administered at a low dose to induce type 2 diabetes (T2D) and at a high dose to induce type 1 diabetes—is commonly used in mice and rats in DFU research. Aim: The objective of this study is to highlight the importance of including male and female rats in DFU research. Methods: Both male and female Sprague-Dawley rats, 6-8 weeks old, were fed a high-fat diet for 9 weeks. STZ (25 mg/kg intraperitoneally) was administered weekly from the 6^th week to induce T2D. Results: Female rats required a higher dose of STZ compared to male rats. The induction of T2D correlated positively with weight gain, which was greater in males than in females. Conclusion: The findings suggest that in addition to gender and weight, other factors may influence the induction of T2D in rats. Most studies in the literature do not use both sexes in DFU research. The distinct responses to STZ and weight gain observed emphasize the need to include both sexes and employ a more detailed approach in preclinical studies to enhance the understanding of DFU wound healing and translate the findings into potential treatments. Relevance for patients: The multifactorial effect on diabetes development, which differs in males and females, suggests the need to consider etiological, physiological, and demographic factors, such as body weight, gender, age, and body mass index, in the prevention and treatment of diabetes. This will also help in planning the individualized treatment for DFUs.

Keywords

Diabetic foot ulcer / Animal model / Type 2 diabetes / Streptozotocin / Gender

Funding

The research was supported by a grant (25270C) from GMS Capitals LLC.

Conflict of interest

Vikrant Rai is an Editorial Board Member of this journal but was not in any way involved in the editorial and peer-review process conducted for this paper, directly or indirectly. Separately, other authors declared that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper.

References

[1] Wang X, Yuan CX, Xu B, Yu Z. Diabetic foot ulcers: Classification, risk factors and management. World J Diabetes 2022; 13:1049-1065. doi: 10.4239/wjd.v13.i12.1049

[2] Volmer-Thole M, Lobmann R. Neuropathy and diabetic foot syndrome. Int J Mol Sci 2016; 17:917. doi: 10.3390/ijms17060917

[3] Armstrong DG, Tan TW, Boulton AJM, Bus SA. Diabetic foot ulcers: A review. JAMA 2023; 330:62-75. doi: 10.1001/jama.2023.10578

[4] Rai V, Le H, Agrawal DK. Novel mediators regulating angiogenesis in diabetic foot ulcer healing. Can J Physiol Pharmacol. 2023; 101:488-501. doi: 10.1139/cjpp-2023-0193

[5] Guo Q, Ying G, Jing O, et al. Influencing factors for the recurrence of diabetic foot ulcers: A meta-analysis. Int Wound J. 2023; 20:1762-1775. doi: 10.1111/iwj.14017

[6] Takahashi Y, Fukusato T. Animal models of liver diseases. In: Animal Models for the Study of Human Disease. Netherlands: Elsevier; 2017. p. 313-339. doi: 10.1016/B978-0-12-809468-6.00013-9

[7] Rai V, Agrawal DK. Male or female sex: Considerations and translational aspects in diabetic foot ulcer research using rodent models. Mol Cell Biochem. 2023; 478:1835-1845. doi: 10.1007/s11010-022-04642-7

[8] Rai V, Moellmer R, Agrawal DK. Clinically relevant experimental rodent models of diabetic foot ulcer. Mol Cell Biochem. 2022; 477:1239-1247. doi: 10.1007/s11010-022-04372-w

[9] Tan MLL, Chin JS, Madden L, Becker DL. Challenges faced in developing an ideal chronic wound model. Expert Opin Drug Discov. 2023; 18:99-114. doi: 10.1080/17460441.2023.2158809

[10] Sanapalli BKR, Yele V, Singh MK, Thaggikuppe Krishnamurthy P, Karri V. Preclinical models of diabetic wound healing: A critical review. Biomed Pharmacother. 2021; 142:111946. doi: 10.1016/j.biopha.2021.111946

[11] Furman BL. Streptozotocin-induced diabetic models in mice and rats. Curr Protoc Pharmacol. 2015; 70:5.47.41-5.47.20. doi: 10.1002/0471141755.ph0547s70

[12] Diaz A, Lopez-Grueso R, Gambini J, et al. Sex differences in age-associated type 2 diabetes in rats-role of estrogens and oxidative stress. Oxid Med Cell Longev. 2019; 2019:6734836. doi: 10.1155/2019/6734836

[13] Yan H, Yang W, Zhou F, et al. 2019; 68:291-304. doi: 10.2337/db18-0638

[14] Pratchayasakul W, Chattipakorn N, Chattipakorn SC. Estrogen restores brain insulin sensitivity in ovariectomized non-obese rats, but not in ovariectomized obese rats. Metabolism. 2014; 63:851-859. doi: 10.1016/j.metabol.2014.03.009

[15] Kawakami M, Yokota-Nakagi N, Uji M, et al. Estrogen replacement enhances insulin-induced AS160 activation and improves insulin sensitivity in ovariectomized rats. Am J Physiol Endocrinol Metab. 2018; 315:E1296-E1304. doi: 10.1152/ajpendo.00131.2018

[16] Pratchayasakul W, Chattipakorn N, Chattipakorn SC. Effects of estrogen in preventing neuronal insulin resistance in hippocampus of obese rats are different between genders. Life Sci. 2011; 89:702-707. doi: 10.1016/j.lfs.2011.08.011

[17] Maric I, Krieger JP, van der Velden P, et al. Sex and species differences in the development of diet-induced obesity and metabolic disturbances in rodents. Front Nutr 2022; 9:828522. doi: 10.3389/fnut.2022.828522

[18] Gheibi S, Kashfi K, Ghasemi A. A practical guide for induction of type-2 diabetes in rat: Incorporating a high-fat diet and streptozotocin. Biomed Pharmacother. 2017; 95:605-613. doi: 10.1016/j.biopha.2017.08.098

[19] Ghasemi A, Jeddi S. Streptozotocin as a tool for induction of rat models of diabetes: A practical guide. EXCLI J. 2023; 22:274-294. doi: 10.17179/excli2022-5720

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

[21] Akinlade OM, Owoyele BV, Soladoye AO. Streptozotocin-induced type 1 and 2 diabetes in rodents: A model for studying diabetic cardiac autonomic neuropathy. Afr Health Sci. 2021; 21:719-727. doi: 10.4314/ahs.v21i2.30

[22] Zhang M, Lv XY, Li J, Xu ZG, Chen L. The characterization of high-fat diet and multiple low-dose streptozotocin induced type 2 diabetes rat model. Exp Diabetes Res. 2008; 2008:704045. doi: 10.1155/2008/704045

[23] Melhorn SJ, Krause EG, Scott KA, et al. Acute exposure to a high-fat diet alters meal patterns and body composition. Physiol Behav. 2010; 99:33-39. doi: 10.1016/j.physbeh.2009.10.004

[24] Uckun FM, Orhan C, Tuzcu M, et al. RJX improves wound healing in diabetic rats. Front Endocrinol (Lausanne). 2022; 13:874291. doi: 10.3389/fendo.2022.874291

[25] Wang J, Zhao X, Tian G, Liu X, Gui C, Xu L. Down-regulation of miR-138 alleviates inflammatory response and promotes wound healing in diabetic foot ulcer rats via activating PI3K/AKT pathway and hTERT. Diabetes Metab Syndr Obes. 2022; 15:1153-1163. doi: 10.2147/DMSO.S359759

[26] Qi L, Ahmadi AR, Huang J, et al. Major improvement in wound healing through pharmacologic mobilization of stem cells in severely diabetic rats. Diabetes. 2020; 69:699-712. doi: 10.2337/db19-0907

[27] Sun X, Wang X, Zhao Z, Chen J, Li C, Zhao G. Paeoniflorin accelerates foot wound healing in diabetic rats though activating the Nrf2 pathway. Acta Histochem. 2020; 122:151649. doi: 10.1016/j.acthis.2020.151649

[28] Cheng KY, Lin ZH, Cheng YP, et al. Wound healing in streptozotocin-induced diabetic rats using atmospheric-pressure argon plasma jet. Sci Rep. 2018; 8:12214. doi: 10.1038/s41598-018-30597-1

[29] Yang P, Pei Q, Yu T, et al. Compromised wound healing in ischemic type 2 diabetic rats. PLoS One. 2016; 11:e0152068. doi: 10.1371/journal.pone.0152068

[30] Karalashvili L, Chakhunashvili D, Kakabadze M, Paresishvili T, Kakabadze Z. Treatment of a non-healing oral wound in diabetic-induced rats. Explor Immunol. 2023; 3:565-573.

[31] Begum F, Manandhar S, Kumar G, et al. Dehydrozingerone promotes healing of diabetic foot ulcers: A molecular insight. J Cell Commun Signal. 2023; 17:673-688. doi: 10.1007/s12079-022-00703-0

[32] Racine KC, Iglesias-Carres L, Herring JA, et al. The high-fat diet and low-dose streptozotocin type-2 diabetes model induces hyperinsulinemia and insulin resistance in male but not female C57BL/6J mice. Nutr Res 2024; 131:135-146. doi: 10.1016/j.nutres.2024.09.008

[33] Kim B, Kim YY, Nguyen PTT, Nam H, Suh JG. Sex differences in glucose metabolism of streptozotocin-induced diabetes inbred mice (C57BL/6J). Appl Biol Chem. 2020; 63:59. doi: 10.1186/s13765-020-00547-5

[34] Kim B, Park ES, Lee JS, Suh JG. Outbred mice with streptozotocin-induced diabetes show sex differences in glucose metabolism. Int J Mol Sci. 2023; 24:5210. doi: 10.3390/ijms24065210

[35] Iwase M, Kikuchi M, Nunoi K, et al. Sex differences in diabetes induced by neonatal streptozotocin treatment in spontaneously hypertensive rats. Diabetes Res Clin Pract. 1987; 4:61-65. doi: 10.1016/s0168-8227(87)80034-7

[36] Tanday N, Coulter-Parkhill A, Moffett RC, et al. Sex-based impact of pancreatic islet stressors in GluCreERT2/Rosa26- eYFP mice. J Endocrinol. 2023; 259:e230174. doi: 10.1530/JOE-23-0174

[37] Sprouse J, Sampath C, Gangula P. 17beta-estradiol suppresses gastric inflammatory and apoptotic stress responses and restores nNOS-mediated gastric emptying in streptozotocin (STZ)-induced diabetic female mice. Antioxidants (Basel). 2023; 12:758. doi: 10.3390/antiox12030758

[38] Tao Z, Cheng Z. Hormonal regulation of metabolism-recent lessons learned from insulin and estrogen. Clin Sci (Lond). 2023; 137:415-434. doi: 10.1042/CS20210519

[39] Gasparini SJ, Swarbrick MM, Kim S, et al. Androgens sensitise mice to glucocorticoid-induced insulin resistance and fat accumulation. Diabetologia. 2019; 62:1463-1477. doi: 10.1007/s00125-019-4887-0

[40] Rehman HU, Ullah K, Rasool A, et al. Comparative impact of streptozotocin on altering normal glucose homeostasis in diabetic rats compared to normoglycemic rats. Sci Rep. 2023; 13:7921. doi: 10.1038/s41598-023-29445-8

[41] Kaler LW, Neaves WB. The androgen status of aging male rats. Endocrinology. 1981; 108:712-719. doi: 10.1210/endo-108-2-712

[42] Turgeon JL, Waring DW. Androgen modulation of luteinizing hormone secretion by female rat gonadotropes. Endocrinology. 1999; 140:1767-1774. doi: 10.1210/endo.140.4.6642

[43] Carpene C, Stiliyanov Atanasov K, Les F, Mercader Barcelo J. Hyperglycemia and reduced adiposity of streptozotocin-induced diabetic mice are not alleviated by oral benzylamine supplementation. World J Diabetes. 2022; 13:752-764. doi: 10.4239/wjd.v13.i9.752

[44] Luo L, Liu M. Adipose tissue in control of metabolism. J Endocrinol. 2016;231:R77-R99. doi: 10.1530/JOE-16-0211

[45] Singla P, Bardoloi A, Parkash AA. Metabolic effects of obesity: A review. World J Diabetes. 2010; 1:76-88. doi: 10.4239/wjd.v1.i3.76

[46] Capponi L, Schmitz A, Thormann W, Theurillat R, Mevissen M. In vitro evaluation of differences in phase 1 metabolism of ketamine and other analgesics among humans, horses, and dogs. Am J Vet Res. 2009; 70:777-786. doi: 10.2460/ajvr.70.6.777

[47] Rebolledo-Solleiro D, Fernandez-Guasti A. Influence of sex and estrous cycle on blood glucose levels, body weight gain, and depressive-like behavior in streptozotocin-induced diabetic rats. Physiol Behav. 2018; 194:560-567. doi: 10.1016/j.physbeh.2018.06.033