Gomisin D alleviates podocyte injury in diabetic kidney disease by inhibiting oxidative stress via activation of the GSK3β (Ser9)/Nrf2/ARE signaling pathway

Abstract

Objective: To explore whether gomisin D, a lignan derived from Schisandra chinensis, protects against podocyte injury in diabetic kidney disease and to elucidate its underlying mechanisms. Methods: Network pharmacology was employed to analyze the potential interactions between gomisin D and podocyte injury-related targets in diabetic nephropathy. A high glucose-induced injury model of mouse podocyte MPC5 cells was established, and the effect of gomisin D on podocyte injury was subsequently evaluated by measurement of reactive oxygen species levels and biochemical markers and Western blotting. Molecular docking and further molecular biology assays were conducted to explore the interaction between gomisin D and the target protein. Results: Gomisin D inhibited high glucose-induced oxidative stress, improved cell viability, and preserved the expression of podocin and synaptopodin. Network pharmacology identified glycogen synthase kinase-3β (GSK3β) as a key target. Molecular docking predicted a potential interaction between gomisin D and GSK3β, which was further validated by drug affinity responsive target stability and cellular thermal shift assay, confirming direct binding in live cells. Mechanistically, gomisin D enhanced the phosphorylation of GSK3β at Ser9, promoted the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2), and upregulated levels of downstream antioxidant proteins heme oxygenase-1 and NAD(P)H quinone oxidoreductase 1. Furthermore, forced expression of the constitutively active GSK3β mutant S9A decreased GSK3β (Ser9) phosphorylation and reversed gomisin D-induced upregulation of these antioxidant markers. Conclusions: Gomisin D mitigates high glucose-induced podocyte injury in vitro by inhibiting oxidative stress via activation of the GSK3β(Ser9)/Nrf2/ARE signaling pathway.

Keywords

Gomisin D / Diabetic kidney disease / Podocyte injury / GSK3β / Nrf2/ARE pathway / Oxidative stress

References

[1] Rayego-Mateos S, Rodrigues-Diez RR, Fernandez-Fernandez B, Mora-Fernández C, Marchant V, Donate-Correa J, et al. Targeting inflammation to treat diabetic kidney disease: The road to 2030. Kidney Int 2023; 103(2): 282-296. doi: 10.1016/j.kint.2022.10.030.

[2] Tan HB, Zhao Q, Chen L. Penehyclidine hydrochloride suppresses inflammation response and reduces podocyte injury in diabetic nephropathy by targeting fibrinogen-like protein 2. Int Immunopharmacol 2022; 107. doi: 10.1016/j.intimp.2022.108680.

[3] Zhang X, Zhang J, Ren Y, Sun R, Zhai X. Unveiling the pathogenesis and therapeutic approaches for diabetic nephropathy: Insights from panvascular diseases. Front Endocrinol (Lausanne) 2024; 15. doi: 10.3389/fendo.2024.1368481.

[4] Li X, Zhang Y, Xing X, Li M, Liu Y, Xu A, et al. Podocyte injury of diabetic nephropathy: Novel mechanism discovery and therapeutic prospects. Biomed Pharmacother 2023; 168. doi: 10.1016/j.biopha.2023.115670.

[5] Tuttle KR, Agarwal R, Alpers CE, Bakris GL, Brosius FC, Kolkhof P, et al. Molecular mechanisms and therapeutic targets for diabetic kidney disease. Kidney Int 2022; 102(2): 248-260. doi: 10.1016/j.kint.2022.05.012.

[6] Liu X, Jiang L, Zeng H, Gao L, Guo S, Chen C, et al. Circ-0000953 deficiency exacerbates podocyte injury and autophagy disorder by targeting Mir665-3p-Atg4b in diabetic nephropathy. Autophagy 2024; 20(5): 1072-1097. doi: 10.1080/15548627.2023.2286128.

[7] Liu XJ, Hu XK, Yang H, Gui LM, Cai ZX, Qi MS, et al. A review of traditional Chinese medicine on treatment of diabetic nephropathy and the involved mechanisms. Am J Chin Med 2022; 50(7): 1739-1779. doi: 10.1142/s0192415x22500744.

[8] Tang G, Li S, Zhang C, Chen H, Wang N, Feng Y. Clinical efficacies, underlying mechanisms and molecular targets of Chinese medicines for diabetic nephropathy treatment and management. Acta Pharm Sin B 2021; 11(9): 2749-2767. doi: 10.1016/j.apsb.2020.12.020.

[9] Peng L, Chen X, Cai H, Tang Y, Chen Q, Zhou F. Wenyang Zhenshuai Granules inhibits cardiomyocyte apoptosis in chronic heart failure by regulating p38 MAPK signaling pathway through exosomal miR-155. Front Pharmacol 2025; 16. doi: 10.3389/fphar.2025.1538091.

[10] Wang JX, Lu Q, Chen QY, Liao Y, Yi Q. Effect of Wenyang Zhenshuai Granules on autophagy and apoptosis of myocardial cells in septic rats via regulating miR-132-3p/UCP2 expression. Zhongguo Zhong Yao Za Zhi 2023; 48(11): 3066-3073. doi: 10.19540/j.cnki.cjcmm.20230216.701.

[11] Szopa A, Dziurka M, Warzecha A, Kubica P, Klimek-Szczykutowicz M, Ekiert H. Targeted lignan profiling and anti-inflammatory properties of Schisandra rubriflora and Schisandra chinensis extracts. Molecules 2018; 23(12): 3103. doi: 10.3390/molecules23123103.

[12] Zhang M, Liu M, Xiong M, Gong J, Tan X. Schisandra chinensis fruit extract attenuates albuminuria and protects podocyte integrity in a mouse model of streptozotocin-induced diabetic nephropathy. J Ethnopharmacol 2012; 141(1): 111-118. doi: 10.1016/j.jep.2012.02.007.

[13] Zhang H, Chen R, Xu C, Zhang G, Guan Y, Feng Q, et al. An integrated approach to discriminate the quality markers of traditional Chinese medicine preparation based on multi-dimensional characteristic network: Shenqi Jiangtang Granule as a case. J Ethnopharmacol 2021; 278. doi: 10.1016/j.jep.2021.114277.

[14] Zheng X, Feng F, Jiang X, Qiu J, Cai X, Xiang Z. A rapid UPLC-MS method for quantification of gomisin D in rat plasma and its application to a pharmacokinetic and bioavailability study. Molecules 2019; 24(7): 1403. doi: 10.3390/molecules24071403.

[15] Sun LN, Liu XC, Chen XJ, Guan GJ, Liu G. Curcumin attenuates high glucose-induced podocyte apoptosis by regulating functional connections between caveolin-1 phosphorylation and ROS. Acta Pharmacol Sin 2016; 37(5): 645-655. doi: 10.1038/aps.2015.159.

[16] Chen M, Fang Y, Ge Y, Qiu S, Dworkin L, Gong R. The redox-sensitive GSK3β is a key regulator of glomerular podocyte injury in type 2 diabetic kidney disease. Redox Biol 2024; 72: 103127. doi: 10.1016/j.redox.2024.103127.

[17] Wang R, Liu F, Chen P, Li S, Gu Y, Wang L, et al. Gomisin D alleviates liver fibrosis through targeting PDGFRβ in hepatic stellate cells. Int J Biol Macromol 2023; 235. doi: 10.1016/j.ijbiomac.2023.123639.

[18] Huang L, Li G, Zhang Y, Zhuge R, Qin S, Qian J, et al. Small-molecule targeting BCAT1-mediated BCAA metabolism inhibits the activation of SHOC2-RAS-ERK to induce apoptosis of Triple-negative breast cancer cells. J Adv Res 2025; 75: 723-738. doi: 10.1016/j.jare.2024.10.021.

[19] Sagoo MK, Gnudi L. Diabetic nephropathy: An overview. Methods Mol Biol 2020; 2067: 3-7. doi: 10.1007/978-1-4939-9841-8_1.

[20] Selby NM, Taal MW. An updated overview of diabetic nephropathy: Diagnosis, prognosis, treatment goals and latest guidelines. Diabetes Obes Metab 2020; 22 (Suppl 1): 3-15. doi: 10.1111/dom.14007.

[21] Martinez Leon V, Hilburg R, Susztak K. Mechanisms of diabetic kidney disease and established and emerging treatments. Nat Rev Endocrinol 2026; 22(1): 21-35. doi: 10.1038/s41574-025-01171-3.

[22] Liu Q, Guo S, Tong Y, Qiu L, Zhuang B, Wang J, et al. Renal lipotoxicity in diabetic and obesity-associated kidney diseases: Molecular mechanisms and therapeutic targeting. Diabetes Obes Metab 2026; 28(1): 60-82. doi: 10.1111/dom.70220.

[23] Zhang M, Tatar M, Gong R. Glycogen synthase kinase 3β: A key player in progressive chronic kidney disease. Clin Sci (Lond) 2025; 139(12): 605-625. doi: 10.1042/cs20245219.

[24] Adelusi TI, Du L, Hao M, Zhou X, Xuan Q, Apu C, et al. Keap1/Nrf2/ARE signaling unfolds therapeutic targets for redox imbalanced-mediated diseases and diabetic nephropathy. Biomed Pharmacother 2020; 123. doi: 10.1016/j.biopha.2019.109732.

[25] Attri M, Kuwar OK. The KEAP1-Cullin3-RBX1-Nrf2 axis in redox homeostasis: Molecular mechanisms, pathophysiological roles, and precision therapeutic opportunities. Mol Neurobiol 2025; 63(1): 15. doi: 10.1007/s12035-025-05313-6.

[26] Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 2009; 284(20): 13291-13295. doi: 10.1074/jbc.R900010200.

[27] Chen L, Chou CL, Knepper MA. A comprehensive map of mRNAs and their isoforms across All 14 renal tubule segments of mouse. J Am Soc Nephrol 2021; 32(4): 897-912. doi: 10.1681/asn.2020101406.

[28] Hurcombe JA, Hartley P, Lay AC, Ni L, Bedford JJ, Leader JP, et al. Podocyte GSK3 is an evolutionarily conserved critical regulator of kidney function. Nat Commun 2019; 10(1): 403. doi: 10.1038/s41467-018-08235-1.

[29] Ahmad F, Gupta A, Marzook H, Woodgett JR, Saleh MA, Qaisar R. Natural compound screening predicts novel GSK-3 isoform-specific inhibitors. Biochimie 2024; 225: 68-80. doi: 10.1016/j.biochi.2024.05.002.

[30] Wang Y, Wang H, Li Y. Protocatechuic aldehyde ameliorates high glucose-induced podocyte injury by attenuating inflammation, oxidative stress, and apoptosis via suppression of endoplasmic reticulum stress through the GSK3β/Nrf2 pathway. Front Cell Dev Biol 2025; 13. doi: 10.3389/fcell.2025.1693955.

[31] Luty RS, Al-Zubaidy AA, Malik AS, Ridha-Salman H, Abbas AH. Protective effect of orientin on diabetic nephropathy in rat models of high-fat diet and streptozotocin-induced diabetes. Naunyn Schmiedebergs Arch Pharmacol 2025; 398(8): 10769-10784. doi: 10.1007/s00210-025-03949-8.

[32] Meng Y, Hou Y, Zhang R, Guo Z, Zhang Z, Li J, et al. Jinlida ameliorates diabetic kidney disease via gut microbiota-dependent production of pyridoxamine targeting renal AGEs/RAGE and TGF-β pathways. Phytomedicine 2026; 150. doi: 10.1016/j.phymed.2025.157744.

[33] Jiang H, Hu XW, Deng X, Huang XJ, Chen YL, Yang YF, et al. Liuwei Dihuang Pills ameliorate renal injury in experimental type 2 diabetes mellitus rat by regulating host-gut microbiota interaction. Front Pharmacol 2025; 16. doi: 10.3389/fphar.2025.1715600.

[34] Huang L, Wu W, Wang X. Analysis of the microecological mechanism of diabetic kidney disease based on the theory of “Gut-Kidney Axis”: A systematic review. Open Life Sci 2024; 19(1). doi: 10.1515/biol-2022-0909.

[35] Jiang H, Wang X, Zhou W, Huang Z, Zhang W. Gut microbiota dysbiosis in diabetic nephropathy: Mechanisms and therapeutic targeting via the gut-kidney axis. Front Endocrinol (Lausanne) 2025; 16. doi: 10.3389/fendo.2025.1661037.

[36] Mei M, Sun H, Zhang K, Zhang F, Sun S, Zhang Y. Phellodendron amurense Rupr. polysaccharides protects against diabetic nephropathy via alteration of PI3K/GSK-3β/Nrf2/TGF-β/Smad signaling pathway and gut microbiota. Nat Prod Bioprospect 2026; 16(1): 13. doi: 10.1007/s13659-025-00566-z.

[37] Yan ZJ, Zhang L, Han XY, Kang Y, Liu SM, Ma TP, et al. Alpiniae oxyphyllae Fructus ameliorates renal lipid accumulation in diabetic kidney disease via activating PPARα. Asian Pac J Trop Biomed 2025; 15(1): 11-23.

[38] Zhou Y, Wei L, Hu L, Han S, Zhang R, Cao Y. Activation of the Lp-PLA2/LPC axis triggers endothelial ferroptosis to drive diabetic kidney disease. Free Radic Biol Med 2025; 241: 818-828. doi: 10.1016/j.freeradbiomed.2025.09.036.

[39] Ma DY, Liu JX, Wang LD, Zhi XY, Luo L, Zhao JY, et al. GSK-3β-dependent Nrf2 antioxidant response modulates ferroptosis of lens epithelial cells in age-related cataract. Free Radic Biol Med 2023; 204: 161-176. doi: 10.1016/j.freeradbiomed.2023.04.022.

[40] Li X, Qian J, Xu J, Bai H, Yang J, Chen L. NRF2 inhibits RSL3 induced ferroptosis in gastric cancer through regulation of AKR1B1. Exp Cell Res 2024; 442(1). doi: 10.1016/j.yexcr.2024.114210.

[41] Duan J, Pei F, Miao J, Liu S, Tan L, Lu M, et al. Swietenine improved the progression of diabetic nephropathy through inhibiting ferroptosis via activating Akt/GSK-3β/Nrf2 signaling pathway. J Ethnopharmacol 2025; 349. doi: 10.1016/j.jep.2025.119981.

[42] Hu J, Zhang Y, Zhang Y, Shi N, Miu Y, Huang J, et al. Bergenin inhibits ferritinophagy and ferroptosis in cisplatin-induced acute kidney injury by activating the p-GSK3β/Nrf2/PPARγ pathway. Int Immunopharmacol 2025; 147. doi: 10.1016/j.intimp.2024.114004.

[43] Zhao W, Jin G, Sun W, Wu C, Yang Q, Xue L, et al. Empagliflozin alleviates type 2 diabetic renal fibrosis by inhibiting SLC7A7-mediated ferroptosis. Diabetol Metab Syndr 2025; 17(1): 329. doi: 10.1186/s13098-025-01902-6.