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中华肾病研究电子杂志

中华肾病研究电子杂志 ›› 2026, Vol. 15 ›› Issue (01) : 15 -20. doi: 10.3877/cma.j.issn.2095-3216.2026.01.003

论著

激活核因子E2相关因子2信号通路抑制高糖诱导人肾小管上皮细胞上皮-间充质转化
赵坤霄, 刘小明, 冯杰, 胡秀红, 赵文婷†(), 林琼真   
  1. 050000　石家庄，河北医科大学第一医院肾内科
  • 收稿日期:2025-07-14 出版日期:2026-02-28
  • 通信作者: 赵文婷
  • 基金资助:
    河北省医学科学研究课题计划(20230992)

Activation of nuclear factor-E2-related factor 2 signaling pathway inhibited high glucose-induced epithelial-mesenchymal transition in human renal tubular epithelial cells

Kunxiao Zhao, Xiaoming Liu, Jie Feng, Xiuhong Hu, Wenting Zhao†(), Qiongzhen Lin   

  1. Department of Nephrology, First Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China
  • Received:2025-07-14 Published:2026-02-28
  • Corresponding author: Wenting Zhao
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引用本文：

赵坤霄, 刘小明, 冯杰, 胡秀红, 赵文婷, 林琼真. 激活核因子E2相关因子2信号通路抑制高糖诱导人肾小管上皮细胞上皮-间充质转化[J/OL]. 中华肾病研究电子杂志, 2026, 15(01): 15-20.

Kunxiao Zhao, Xiaoming Liu, Jie Feng, Xiuhong Hu, Wenting Zhao, Qiongzhen Lin. Activation of nuclear factor-E2-related factor 2 signaling pathway inhibited high glucose-induced epithelial-mesenchymal transition in human renal tubular epithelial cells[J/OL]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2026, 15(01): 15-20.

目的

探讨核因子E2相关因子2(nuclear factor-E2-related factor 2，Nrf2)在高糖诱导人肾小管上皮细胞(human kidney-2, HK-2)间充质转化中的作用。

方法

培养的HK-2细胞分组：正常糖组(NG组，5.5 mmol/L葡萄糖)、高糖组(HG组，30 mmol/L葡萄糖)、高糖加Nrf2 siRNA转染组(HG＋siNrf2组)、高糖加阴性对照siRNA转染组(HG＋NC组)、高糖加N-乙酰半胱氨酸组(HG＋NAC组)、高糖加叔丁基对苯二酚组(HG＋TBHQ组)。应用Western印迹、实时荧光定量PCR和免疫荧光方法检测α-平滑肌肌动蛋白、纤维连接蛋白等表达变化，使用试剂盒检测细胞及线粒体活性氧水平。

结果

相较于NG组，HG组的α-平滑肌肌动蛋白表达较高。与HG组相比，HG＋TBHQ组、HG＋NAC组的α-平滑肌肌动蛋白、纤维连接蛋白的蛋白及mRNA表达则均降低(P均<0.05)；与HG＋NC组相比，HG＋siNrf2组的α-平滑肌肌动蛋白、纤维连接蛋白的蛋白及mRNA表达均较高(P均<0.05)。与NG组相比，HG组的细胞和线粒体活性氧水平均较高(P均<0.05)；与HG组相比，HG＋TBHQ组和HG＋NAC组的细胞和线粒体活性氧水平均较低(P均<0.05)。

结论

激活Nrf2信号通路能够抑制高糖诱导人肾小管上皮细胞的活性氧生成和上皮-间充质转化。

关键词: 高糖, 人肾小管上皮细胞, 氧化应激, 核因子E2相关因子2, 上皮-间充质转化
Objective

To investigate the role of nuclear factor-E2-related factor 2 (Nrf2) in high glucose-induced epithelial-mesenchymal transition (EMT) in human renal tubular epithelial cells (human kidney-2, HK-2).

Methods

Cultured HK-2 cells were divided into: normal glucose group (NG group, 5.5 mmol/L glucose), high glucose group (HG group, 30 mmol/L glucose), high glucose plus Nrf2 siRNA transfection group (HG+ siNrf2 group), high glucose plus negative control siRNA transfection group (HG+ NC group), high glucose plus N-acetylcysteine group (HG+ NAC group), and high glucose plus tert-butylhydroquinone group (HG+ TBHQ group). The expression levels of α-smooth muscle actin and fibronectin were examined using Western blotting, quantitative real-time PCR, and immunofluorescence. Intracellular and mitochondrial reactive oxygen species (ROS) levels were measured with specific assay kits.

Results

Compared with the NG group, α-smooth muscle actin expression was significantly upregulated in the HG group. In contrast, the protein and mRNA expression levels of α-smooth muscle actin and fibronectin were markedly downregulated in the HG+ TBHQ and HG+ NAC groups compared to the HG group (all P<0.05). Furthermore, both the protein and mRNA expression levels of α-smooth muscle actin and fibronectin were significantly higher in the HG+ siNrf2 group than in the HG+ NC group (all P<0.05). Compared with the NG group, both intracellular and mitochondrial ROS levels were significantly higher in the HG group (all P<0.05). In contrast, the levels of intracellular and mitochondrial ROS were markedly lower in the HG+ TBHQ and HG+ NAC groups compared to the HG group (all P<0.05).

Conclusion

Activation of the Nrf2 signaling pathway inhibited high glucose-induced ROS production and EMT in human renal tubular epithelial cells.

Key words: High glucose, Human renal tubular epithelial cells, Oxidative stress, Nuclear factor-E2-related factor 2, Epithelial-mesenchymal transition
表1 引物序列
检测指标 上游引物序列(5′-3′) 下游引物序列(5′-3′)
Nrf2 5′-TCAGCG ACGGAAAGAGTATGA-3′ 5′-CCACTGGTTTCTGACTGGATGT-3′
α-SMA 5′-TGCACAAACGAGGGGAATATAT-3′ 5′-CGTTCCTTGTAGCCAATAAAGG-3′
FN 5′-TGGACCAAATCTCCACGGTCT-3′ 5′-CTCTCCCCAGGTGATGCCTT-3′
β-actin 5′-TGACGTGGACATCCGCAAAG-3′ 5′-CTGGAAGGTGGACAGCGAGG-3′
图1 高糖对HK-2细胞Nrf2信号通路相关因子的影响注：Nrf2: nuclear factor-E2-related factor 2,核因子E2相关因子2；HO-1: hemeoxygenase-1,血红素氧合酶-1；β-肌动蛋白为内参照；A、B和C：Western印迹法检测各组HK-2细胞中Nrf2、HO-1蛋白表达及比较；D：qPCR法检测各组HK-2细胞中Nrf2 mRNA表达水平；与NG组相比，aP<0.05
图2 高糖对HK-2细胞上皮-间充质转化相关标志物表达的影响注：α-SMA: α-smooth muscle actin，α-平滑肌肌动蛋白；p-Smad3：磷酸化Smad3；TGF-β1: transforming growth factor-β1，转化生长因子-β1；FN: fibronectin,纤维连接蛋白；A：Western印迹法检测各组HK-2细胞中α-SMA、p-Smad3、TGF-β1、FN蛋白表达(β-肌动蛋白为内参照)；B、C、D和E：各组HK-2细胞中α-SMA、p-Smad3、TGF-β1、FN蛋白相对表达的比较；F和G：qPCR法检测各组HK-2细胞中α-SMA的mRNA表达水平；H：免疫荧光法检测HK-2细胞中α-SMA、TGF-β1的表达(绿色荧光×400)；与NG组相比，aP<0.05