Diabetic Nephropathy Treatment by Zn Supplementation in a Murine Model of Type 1 Diabetes Mellitus: Potential Role of Nrf2 Signaling Pathway

XU Lianhong, SUN Weixia, LIU Wei, LI Yanbo

Abstract

To evaluate the renoprotective effects of zinc (Zn) supplementation in diabetes kidney disease (DKD) and to explore its impact on the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway.

Methods 

 A total of 12 male OVE26 mice (spontaneous type 1 diabetes mellitus mice) aged 3 months and weighing approximately 24-27 g were selected and randomly assigned to a diabetes mellitus (DM) group and a zinc-treated DM (DM/Zn) group (n = 6 each). In addition, 12 age-matched male FVB mice weighing approximately 27-30 g were selected and randomly assigned to a non-diabetic control (Ctrl) group and a zinc-treated (Zn) group (n = 6 each). Mice in the DM/Zn and Zn groups were given zinc supplementation for 3 months, with each mouse receiving 5 mg/kg of zinc sulfate by gavage every other day. Mice in the DM and Ctrl groups were given the same volume of normal saline. At the end of the experiment, the albumin-to-creatinine ratio (ACR) in urine was used as an indicator to evaluate renal function. Sirius red staining was performed to assess renal fibrosis in each group of mice. Western blotting was performed to determine the expression of fibrotic growth factors, including connective tissue growth factor (CTGF) and transforming growth factor-β1 (TGF-β1), in renal tissue, and the protein expression of Nrf2, an antioxidant substance, and the protein expression levels of its downstream targets, including NAD(P)H quinone dehydrogenase 1 (NQO1), heme oxygenase 1 (HO-1), superoxide dismutase (SOD)-1, SOD-2, and catalase (CAT).

Results 

1) Compared to the Ctrl group, the urinary protein secretion levels of mice in the DM group exhibited progressive increase. After 3 months of zinc supplementation treatment, the urinary protein secretion levels of mice in the DM/Zn group decreased Compared to that of mice in the DM group, and the difference was statistically significant (P < 0.05). 2) Compared to that in the Ctrl group, the collagen deposition in the renal tissues of mice in the DM group increased, and the difference was statistically significant (P < 0.05), while no obvious change was observed in mice in the DM/Zn group. Compared to the Ctrl group, mice in the DM group exhibited increased expression levels of CTGF and TGF-β1 in the renal tissues, but the expression levels decreased after zinc supplementation treatment, with the differences being statistically significant (P < 0.05). 3) Compared to that of the Ctrl group, the expression level of Nrf2 in the renal tissues of mice in the Zn and DM groups increased, and the level of Nrf2 in the renal tissues of mice in the DM/Zn group showed a further increase, with the differences being statistically significant (P < 0.05). 4) Compared to those of the Ctrl group, the protein expression levels of Nrf2 downstream target genes, including NQO1 and HO-1, in the renal tissues of mice in the Zn group increased, and the levels of NQO1 and HO-1 in the renal tissues of mice in the DM/Zn group showed a further increase, with the differences being statistically significant (P < 0.05). Compared to those of the mice in the Ctrl group, the protein expressions of Nrf2 downstream target genes, including SOD-1, SOD-2, and CAT of in the renal tissues of the mice in the Zn group increased, while the expression levels of SOD-1, SOD-2, and CAT in the renal tissues of the mice in the DM group decreased, with the differences being statistically significant (P < 0.05). Zn supplementation could completely inhibit these changes (P < 0.05).

Conclusions 

Zn supplementation has therapeutic effects on DKD and mitigates T1DM-induced renal dysfunction and oxidative injury in mice, which may be associated with the activation of the Nrf2 antioxidant signaling pathway.

 

Keywords: Zn, Diabetic nephropathy, Antioxidant substance, Nrf2

 

Full Text:

PDF


References


JIN Q, LIU T, QIAO Y, et al. Oxidative stress and inflammation in diabetic nephropathy: role of polyphenols. Front Immunol, 2023, 14: 1185317. doi: 10.3389/fimmu.2023.1185317.

ABD EL-KHALIK S R, NASIF E, ARAKEEP H M, et al. The prospective ameliorative role of zinc oxide nanoparticles in STZ-induced diabetic nephropathy in rats: Mechanistic targeting of autophagy and regulating Nrf2/TXNIP/NLRP3 inflammasome signaling. Biol Trace Elem Res, 2022, 200(4): 1677-1687. doi: 10.1007/s12011-021-02773-4.

OTHMAN M S, HAFEZ M M, ABDEL MONEIM A E. The potential role of zinc oxide nanoparticles in microRNAs dysregulation in STZ-induced type 2 diabetes in rats. Biol Trace Elem Res, 2019, 197(2): 606-618. doi: 10. 1007/s12011-019-02012-x.

HOSSEINI R, MONTAZERIFAR F, SHAHRAKI E, et al. The effects of zinc sulfate supplementation on serum copeptin, C-reactive protein and metabolic markers in zinc-deficient diabetic patients on hemodialysis: A randomized, double-blind, placebo-controlled trial. Biol Trace Elem Res, 2022, 200(1): 76-83. doi: 10.1007/s12011-021-02649-7.

WU H, KONG L, TAN Y, et al. C66 ameliorates diabetic nephropathy in mice by both upregulating NRF2 function via increase in miR-200a and inhibiting miR-21. Diabetologia, 2016, 59(7): 1558-1568. doi: 10.1007/s00125-016-3958-8.

NIE P, LOU Y, BAI X, et al. Influence of zinc levels and Nrf2 expression in the clinical and pathological changes in patients with diabetic nephropathy. Nutr Diabetes, 2022, 12(1): 37. doi: 10.1038/s41387-022-00212-4.

AHMED MUSTAFA Z, HAMED Ali R, ROSTUM Ali D, et al. The combination of ginger powder and zinc supplement improves the fructose-induced metabolic syndrome in rats by modulating the hepatic expression of NF-kappaB, mTORC1, PPAR-alpha SREBP-1c, and Nrf2. J Food Biochem, 2021, 45(1): e13546. doi: 10.1111/jfbc.13546.

ZHENG S, NOONAN W T, METREVELI N S, et al. Development of late-stage diabetic nephropathy in OVE26 diabetic mice. Diabetes, 2004, 53(12): 3248-3257. doi: 10.2337/diabetes.53.12.3248.

WANG J, SONG Y, ELSHERIF L, et al. Cardiac metallothionein induction plays the major role in the prevention of diabetic cardiomyopathy by zinc supplementation. Circulation, 2006, 113(4): 544-554. doi: 10.1161/CIRCULATIONAHA.105.537894.

LIN Q Y, LI M J, LIANG D, et al. Research progress of Nrf2 in oxidative stress-induced renal fibrosis in diabetic kidney disease. Chin J Diabetes, 2023, 31(8): 627-631. doi: 10.3969/j.issn.1006-6187.2023.08.013.

SUN H J, WU Z Y, CAO L, et al. Hydrogen sulfide: recent progression and perspectives for the treatment of diabetic nephropathy. Molecules, 2019, 24(15): 2857. doi: 10.3390/molecules24152857.

BAPPUTTY R, TALAHALLI R, ZARINI S, et al. Montelukast prevents early diabetic retinopathy in mice. Diabetes, 2019, 68(10): 2004-2015. doi: 10.2337/db19-0026.

JIN B, CHEN Y, WANG J, et al. Costunolide alleviates hyperglycaemia-induced diabetic cardiomyopathy via inhibiting inflammatory responses and oxidative stress. J Cell Mol Med, 2023, 27(6): 831-845. doi: 10.1111/jcmm.17686.

ROBERTSON R P. Nrf2 and antioxidant response in animal models of type 2 diabetes. Int J Mol Sci, 2023, 24(4): 3082. doi: 10.3390/ijms24043082.

DODSON M, SHAKYA A, ANANDHAN A, et al. NRF2 and diabetes: The good, the bad, and the complex. Diabetes, 2022, 71(12): 2463-2476. doi: 10.2337/db22-0623.

NEGI C K, JENA G. Jena, Nrf2, a novel molecular target to reduce type 1 diabetes associated secondary complications: the basic considerations. Eur J Pharmacol, 2019, 843: 12-26. doi: 10.1016/j.ejphar.2018.10.026.

DING Q, SUN B, WANG M, et al. N-acetylcysteine alleviates oxidative stress and apoptosis and prevents skeletal muscle atrophy in type 1 diabetes mellitus through the NRF2/HO-1 pathway. Life Sci, 2023, 329: 121975. doi: 10.1016/j.lfs.2023.121975.

TAN Y, ICHIKAWA T, LI J, et al. Diabetic downregulation of Nrf2 activity via ERK contributes to oxidative stress-induced insulin resistance in cardiac cells in vitro and in vivo. Diabetes, 2011, 60(2): 625-633. doi: 10. 2337/db10-1164.

LIU X, CHEN C H. Advances of nuclear factor erythroid 2 related factor 2 regulation of cell homeostasis in diabetic kidney disease. J Nephrol Dialy Transplant, 2024, 33(2): 155-159. doi: 10.3969/j.issn.1006-298X.2024.02.011.

YANG F, LI B, DONG X, et al. The beneficial effects of zinc on diabetes-induced kidney damage in murine rodent model of type 1 diabetes mellitus. J Trace Elem Med Biol, 2017, 42: 1-10. doi: 10.1016/j.jtemb.2017. 03.006.

LV Y, JIANG H, LI S, et al. Sulforaphane prevents chromium-induced lung injury in rats via activation of the Akt/GSK-3beta/Fyn pathway. Environ Pollut, 2020, 259: 113812. doi: 10.1016/j.envpol.2019.113812.

MANCUSO F, ARATO I, BELLUCCI C, et al. Zinc restores functionality in porcine prepubertal Sertoli cells exposed to subtoxic cadmium concentration via regulating the Nrf2 signaling pathway. Front Endocrinol (Lausanne), 2023, 14: 962519. doi: 10.3389/fendo.2023. 962519.


Refbacks

  • There are currently no refbacks.