二甲双胍通过上调Nrf2表达抑制糖尿病肾小管上皮细胞铁死亡

doi:10.12092/j.issn.1009-2501.2026.01.004

摘要/Abstract

摘要：

目的: 探究二甲双胍通过促进核因子E2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）表达抑制糖尿病肾小管上皮细胞铁死亡的作用机制。方法: 8周龄DBA2/J雄性小鼠分为正常对照组、单纯二甲双胍（metformin，Met）组、糖尿病（DM）组、糖尿病（DM）+二甲双胍（Met）治疗组，每组6只。人肾小管上皮细胞（HK-2）被分为正常对照（5.5 mmmol/L，Ctrl）组、二甲双胍（Met）组、高糖（30 mmmol/L，HG）组和高糖+二甲双胍（HG+Met）组共4组。测定各组小鼠血肌酐、尿素氮，观察肾脏组织病理改变，检测各组肾脏组织和HK-2细胞的活性氧（ROS）变化、Nrf2、GPX4、FTH-1的蛋白和mRNA表达水平，并测定丙二醛（MDA）、铁离子（Fe²⁺）和谷胱甘肽（GSH）的含量。结果: 与DM组相比，DM+Met组中二甲双胍可显著降低糖尿病小鼠的血清尿素氮、血清肌酐水平和肾小管损伤评分（均P<0.05），显著提高糖尿病小鼠肾脏组织 Nrf2、GPX4及FTH-1蛋白表达（均P<0.05）。与HG组相比，二甲双胍能够显著促进高糖诱导HK-2细胞内Nrf2、GPX4、FTH-1蛋白和mRNA的表达量，且显著增加GSH含量，并显著减少细胞内ROS、MDA和Fe²⁺含量（均P<0.05）。结论: 二甲双胍可抑制高糖诱导的HK-2细胞铁死亡，从而缓解糖尿病肾小管病变，其机制可能是通过上调肾小管上皮细胞内Nrf2的表达。

关键词: 二甲双胍, 铁死亡, 糖尿病, Nrf2

Abstract:

AIM: To investigate the mechanism of metformin in inhibiting ferroptosis of diabetic renal tubular epithelial cells by promoting the expression of Nrf2. METHODS: 8-week-old DBA2/J male mice were divided into normal control group, Met group, DM group and DM+Met group. HK-2 cells were divided into four groups: normal glucose group, Met group, high glucose group (30 mmmol/L) and HG+Met group. The changes of Scr, BUN, ROS, the protein and mRNA expression levels of Nrf2, GPX4 and FTH-1 were measured in the kidney of mice or HK-2 cells. The contents of MDA, Fe²⁺ and GSH were also detected in renal tissues or the HK-2 cells. RESULTS: Compared with the DM group, the levels of BUN, Scr and the score of renal pathological injury scores were significantly reduced and the protein expressions of Nrf2, GPX4 and FTH-1 were significantly increased in the kidney tissue of diabetes mice treated with metformin (all P<0.05). Compared with the HG group, the expression levels of Nrf2, GPX4, FTH-1 protein and mRNA were significantly promoted, the GSH content was increased and intracellular ROS, MDA, and Fe²⁺ content were decreased in high glucose-induced HK-2 cells treated with metformin (all P<0.05). CONCLUSION: Metformin can suppress ferroptosis induced by high glucose and alleviate diabetic tubulopathy. The mechanism may be through up-regulating the expression of Nrf2 in renal tubular epithelial cells.

Key words: metformin, ferroptosis, diabetes, Nrf2

中图分类号:

吴子瑜, 俞婷, 郑谋炜, 郭太林. 二甲双胍通过上调Nrf2表达抑制糖尿病肾小管上皮细胞铁死亡[J]. 中国临床药理学与治疗学, 2026, 31(1): 40-47.

Ziyu WU, Ting YU, Mouwei ZHENG, Tailin GUO. Metformin inhibits diabetes induced ferroptosis in renal proximal tubular epithelial cells by up-regulating Nrf2 expression[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(1): 40-47.

图/表 5

图 1

Fig.1 Protective effect of metformin on renal tubular injury in diabetic mice ($ \overline{x} $±s, n=6) A: Serunm ceatinine of the mice; B: Serunm urea nitrogen of the mice; C: HE staining of renal tissue (×200); D: Renal tubular pathological injury score.

图 2

Fig.2 Metformin activates Nrf2 expression in the kidney of diabetic mice ($ \overline{x} $±s, n=6) A: the expression of nuclear factor e2-related factor 2 (Nrf2) protein (immunohistochemistry, ×200); B: the expression of Nrf2 protein (Western blot) in kidney tissue; C: the expression of Nrf2 mRNA in kidney tissue.

图 3

Fig.3 Metformin alleviates ferroptosis in the kidney of diabetic mice by inncreasing the expression of GPX4/FTH-1 ($ \overline{x} $±s, n=3) A: Expression of GPX4 and FTH-1 protein in renal tissue (Western blotting); B: The expression of GPX4 mRNA in renal tissue; C: The expression of FTH-1 mRNA in renal tissue; D: The GSH content in kidney tissue; E: The MDA content in kidney tissue; E: The iron content in kidney tissue.

图 4

Fig.4 The Nrf2/GPX4/FTH-1 pathway was activated by metformin in HK-2 cells induced high glucose ($ \overline{x} $±s, n=3) A: The expression of Nrf2, GPX4, FTH-1 protein in HK-2 cells were detected by western blotting; B: The expression of Nrf2 mRNA in HK-2 cells; C: The expression of GPX4 mRNA in HK-2 cells; D: The expression of FTH-1 mRNA in HK-2 cells.

图 5

Fig.5 The metformin reduced reactive oxygen species production and ferroptosis induced by high glucose in HK-2 cells ($ \overline{x} $±s, n=3) A: Changes of reactive oxygen species in HK-2 cells; B: The GSH content in HK-2 cells; C: The MDA content in HK-2 cells; D: The iron content in HK-2 cells.

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