中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (11): 2774-2783.doi: 10.12307/2026.124

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

灰兜巴抑制高糖培养HK-2细胞的铁死亡减轻细胞纤维化

姚顺华，黄彩丁，张梦玉，张可馨，尹长江，杨坤宝   

  1. 承德医学院中药研究所，河北省承德市   067000
  • 收稿日期:2025-03-06 接受日期:2025-07-04 出版日期:2026-04-18 发布日期:2025-09-05
  • 通讯作者: 杨坤宝，博士，教授，承德医学院中药研究所，河北省承德市 067000 并列通讯作者：尹长江，硕士，副教授，承德医学院中药研究所，河北省承德市 067000
  • 作者简介:姚顺华，男，1999年生，山西省晋中市人，汉族，承德医学院在读硕士，主要从事中药、民族药治疗糖尿病及其并发症作用机制研究。
  • 基金资助:
    国家自然科学基金面上项目(82474675)，项目负责人：杨坤宝；国家自然科学基金青年基金项目(81804217)，项目负责人：杨坤宝；河北省人社厅三三三人才项目(C20231059)，项目负责人：杨坤宝

Huidouba inhibits ferroptosis in high glucose-cultured HK-2 cells to attenuate cell fibrosis

Yao Shunhua, Huang Caiding, Zhang Mengyu, Zhang Kexin, Yin Changjiang, Yang Kunbao   

  1. Institute of Traditional Chinese Medicine, Chengde Medical University, Chengde 067000, Hebei Province, China
  • Received:2025-03-06 Accepted:2025-07-04 Online:2026-04-18 Published:2025-09-05
  • Contact: Yang Kunbao, PhD, Professor, Institute of Traditional Chinese Medicine, Chengde Medical University, Chengde 067000, Hebei Province, China Co-corresponding author: Yin Changjiang, MS, Associate professor, Institute of Traditional Chinese Medicine, Chengde Medical University, Chengde 067000, Hebei Province, China
  • About author:Yao Shunhua, MS candidate, Institute of Traditional Chinese Medicine, Chengde Medical University, Chengde 067000, Hebei Province, China
  • Supported by:
    the National Natural Science Foundation of China (General Program), No. 82474675 (to YKB); National Natural Science Foundation of China for the Youth, No. 81804217 (to YKB); “333 Talent Project” of Hebei Provincial Department of Human Resources and Social Affairs, No. C20231059 (to YKB) 

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摘要：


文题释义：
糖尿病肾病：是糖尿病最常见的微血管并发症，其发病机制复杂，涉及遗传、代谢紊乱、氧化应激、炎症反应等。
铁死亡：是一种由铁过载和脂质过氧化引起的非调节性细胞死亡形式，主要表现为对氧化应激的防御降低和异常的脂质过氧化，当细胞内的抗氧化系统被抑制时，无法清除过多的脂质过氧化产物，引起细胞膜损伤，最终诱导细胞发生铁死亡。

背景：糖尿病肾病与铁死亡密切相关，既往研究发现灰兜巴具有显著的降糖作用，对糖尿病肾病引起的肾脏氧化应激损伤及纤维化具有保护作用，还可降低高糖诱导的HK-2细胞纤维化水平。
目的：探究灰兜巴调控谷胱甘肽过氧化物酶4(Glutathione Peroxidase 4，GPX4)抑制高糖培养人肾皮质近曲小管上皮细胞(HK-2)铁死亡减轻细胞纤维化作用。
方法：①为探究灰兜巴对高糖培养HK-2细胞铁死亡及纤维化的干预作用，将细胞分为6组：对照组、模型组、灰兜巴低、中、高剂量组、罗格列酮组；观察HK-2细胞形态，检测细胞存活率及氧化应激相关指标丙二醛含量、谷胱甘肽水平，检测铁死亡相关指标GPX4、铁蛋白重链1、核因子E2相关因2、转铁蛋白受体及纤维化相关指标转化生长因子β1、肾损伤分子1的蛋白和mRNA表达。②为观察siRNA干扰HK-2细胞GPX4表达效果，将细胞分为NC组(阴性对照)、si-GPX4组；检测GPX4蛋白和mRNA表达。③为观察siRNA干扰GPX4对灰兜巴抑制高糖培养HK-2细胞铁死亡减轻细胞纤维化作用效果的影响，将细胞分为4组：NC组(阴性对照)、高糖组(30 mmol/L葡萄糖处理)、高糖+灰兜巴组 (30 mmol/L葡萄糖+400 μg/mL灰兜巴水提物处理)、高糖+灰兜巴+si-GPX4组(经GPX4-siRNA处理后，再30 mmol/L葡萄糖+400 μg/mL灰兜巴水提物处理)；检测丙二醛含量、谷胱甘肽水平、GPX4、铁蛋白重链1、核因子E2相关因2、转铁蛋白受体、转化生长因子β1、肾损伤分子1的蛋白和mRNA表达。
结果与结论：①与对照组相比，模型组细胞存活率、谷胱甘肽水平、GPX4、铁蛋白重链1、核因子E2相关因2蛋白和mRNA表达显著降低，丙二醛含量、转铁蛋白受体、转化生长因子β1、肾损伤分子1蛋白和mRNA表达显著升高；与模型组相比，灰兜巴各剂量及罗格列酮组可不同程度改善上述指标变化。②与NC组相比，si-GPX4组GPX4蛋白和mRNA表达显著降低；与高糖+灰兜巴组相比，高糖+灰兜巴+si-GPX4组谷胱甘肽水平、GPX4、铁蛋白重链1、核因子E2相关因2蛋白和mRNA表达显著降低，丙二醛含量、转铁蛋白受体、转化生长因子β1、肾损伤分子1蛋白和mRNA表达显著升高。③结果说明，灰兜巴可通过上调GPX4抑制高糖培养HK-2细胞铁死亡减轻细胞纤维化。                                     
https://orcid.org/0009-0005-2518-3477 (姚顺华) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 糖尿病肾病, 纤维化, HK-2细胞, 铁死亡, 灰兜巴, GPX4

Abstract: BACKGROUND: Diabetic nephropathy is closely related to ferroptosis. Previous studies have found that Huidouba has a significant hypoglycemic effect, protects against diabetic nephropathy-induced renal oxidative stress injury and fibrosis, and also reduces the level of fibrosis in high glucose-induced HK-2 cells. 
OBJECTIVE: To investigate the role of Huidouba in regulating glutathione peroxidase 4 (GPX4) to inhibit ferroptosis and attenuate cellular fibrosis in human renal cortical proximal tubular epithelial cells (HK-2) cultured in high glucose conditions.
METHODS: (1) In order to investigate the interventional effect of Huidouba on ferroptosis and fibrosis in high glucose-cultured HK-2 cells, the cells were divided into six groups: control group, model group, low-, medium-, high-dose Huidouba groups, and rosiglitazone group. The morphology of HK-2 cells was observed. Cell viability, levels of oxidative stress-related indicators (malondialdehyde and glutathione), and protein and mRNA expression levels of ferroptosis-related indicators (GPX4, ferritin heavy chain 1, nuclear factor-E2-associated factor 2, transferrin receptor) and fibrosis-related indicators (transformed growth factor-β1 and kidney injury molecule 1), were detected. (2) To observe the effect of siRNA interfering with GPX4 expression in HK-2 cells, the cells were divided into negative control group and si-GPX4 group. The expression of GPX4 at protein and mRNA levels was detected. (3) In order to observe the effect of siRNA interference with GPX4 on the effect of Huidouba on the inhibition of ferroptosis and mitigation of cellular fibrosis in high glucose-cultured HK-2 cells, the cells were divided into negative control group, high glucose group (treated with 30 mmol/L glucose), high glucose+Huidouba group (treated with 30 mmol/L glucose+400 μg/mL Huidouba water extract), and high glucose+Huidouba+si-GPX4 group (treated with GPX4-siRNA followed by intervention with 30 mmol/L glucose+400 μg/mL Huidouba water extract). The levels of malondialdehyde and glutathione, and the protein and mRNA expression of GPX4, ferritin heavy chain 1, nuclear factor-E2-associated factor 2, transferrin receptor, transformed growth factor-β1, and kidney injury molecule 1 were detected. 
RESULTS AND CONCLUSION: (1) Compared with the control group, the cell viability, glutathione level, the protein and mRNA expression of GPX4, ferritin heavy chain 1, and nuclear factor-E2-associated factor 2 were significantly reduced in the model group, while the malondialdehyde level and protein and mRNA expression of transferrin receptor, transformed growth factor-β1, and kidney injury molecule 1 were significantly increased. Compared with the model group, the above indicators were improved to varying degrees in different dose groups of Huidouba and rosiglitazone group. (2) The protein and mRNA expression of GPX4 was significantly lower in the si-GPX4 group compared with the negative control group. Compared with the high glucose+Huidouba group, the glutathione level, the protein and mRNA expression of GPX4, ferritin heavy chain 1, and nuclear factor-E2-associated factor 2 were significantly reduced in the high glucose+Huidouba+si-GPX4 group, while the malondialdehyde level and protein and mRNA expression of transferrin receptor, transformed growth factor-β1, and kidney injury molecule 1 were significantly increased. To conclude, Huidouba attenuates cell fibrosis by inhibiting ferroptosis in high glucose-cultured HK-2 cells through the upregulation of GPX4.


Key words: diabetic nephropathy, fibrosis, HK-2 cells, ferroptosis, Huidouba, GPX4

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