微小RNA-26a通过抑制铁死亡减少高糖诱导的肾小管上皮细胞细胞外基质合成
microRNA-26a inhibits extracellular matrix synthesis in high glucose-induced renal tubular epithelial cells by regulating ferroptosis
李星月 1乔云阳 1郑辉 1季嘉玲 1张爱青2
作者信息
- 1. 南京医科大学第二附属医院儿科,南京 210003
- 2. 南京医科大学第四附属医院儿科,南京 210031
- 折叠
摘要
目的 探究微小RNA-26a(miR-26a)对高糖(HG)诱导的肾小管上皮细胞(RTECs)细胞外基质(ECM)合成的作用及可能机制.方法 通过HG诱导RTECs以构建糖尿病肾病(DKD)模型,在HG诱导的RTECs中过表达miR-26a,使用RT-qPCR和Western blot检测ECM合成及铁死亡相关指标以评估miR-26a对HG诱导的RTECs中ECM合成及铁死亡的作用.使用ferrostatin(Fer-1)抑制DKD模型中铁死亡的发生并进一步评估其对ECM合成的影响.RT-qPCR和Western blot检测铁死亡的相关指标,荧光显微镜观察活性氧(ROS)荧光强度.结果 与Control相比,HG组细胞中miR-26a表达量降低,ECM合成相关指标fibronectin和colla-gen Ⅰ表达量升高,过表达miR-26a后,与HG组相比,HG+miR-26a 组细胞 miR-26a 表达量升高,fibronectin 和 collagenⅠ表达量降低.在铁死亡方面,与Control组相比,HG组SLC7A11和GPX4的蛋白和mRNA表达量降低,TFR-1和ACSL4表达量升高,ROS荧光强度增强.抑制铁死亡后,HG+Fer-1组铁死亡及ECM合成相关指标表达水平较HG组均改变.再次过表达miR-26a后,HG+miR-26a组铁死亡相关指标表达水平较HG组均变化,ROS荧光强度降低.结论 在HG诱导的RTECs中,miR-26a抑制铁死亡的发生进而减少ECM合成.
Abstract
Objective To investigate the effect and possible mechanism of microRNA-26a(miR-26a)on the syn-thesis of extracellular matrix(ECM)induced by high glucose(HG)in renal tubular epithelial cells(RTECs).Methods A model of diabetic kidney disease(DKD)was constructed by inducing RTECs with HG.MiR-26a was overexpressed in HG-induced RTECs,and RT-qPCR and Western blot were used to assess the effects of miR-26a on ECM synthesis and ferroptosis-related markers in HG-treated RTECs.Ferrostatin(Fer-1)was used to inhibit ferroptosis in the DKD model,and its impact on ECM synthesis was evaluated.RT-qPCR and Western blot were performed to measure ferroptosis-related markers,and fluorescence microscopy was used to observe the intensity of reactive oxygen species(ROS).Results Compared with the control group,the expression of miR-26a decreased in HG-treated cells,while the expression levels of ECM synthesis-related indexes fibronectin and collagen Ⅰ in-creased.After overexpressing miR-26a,the HG+miR-26a group showed a significant increase in miR-26a expres-sion and a decrease in fibronectin and collagen Ⅰ expression compared to the HG group.In terms of ferroptosis,the protein and mRNA expression of SLC7A11 and GPX4 significantly decreased,the expression of TFR-1 and AC-SL4 significantly increased,and the fluorescence intensity of ROS was significantly enhanced in the HG group com-pared with the control group.Inhibition of ferroptosis in the HG+Fer-1 group resulted in significant changes in fer-roptosis and ECM synthesis-related indicators expression levels compared to the HG group.Furthermore,re-expres-sion of miR-26a in the HG+miR-26a led to significant changes in ferroptosis-related indicators expression levels and decreased ROS fluorescence intensity compared to the HG group.Conclusions In HG-induced RTECs,miR-26a inhibits the occurrence of ferroptosis,thus reducing ECM synthesis.
关键词
微小RNA-26a/高糖/肾小管上皮细胞/细胞外基质/铁死亡Key words
microRNA-26a/high glucose/renal tubular epithelial cells/extracellular matrix/ferroptosis引用本文复制引用
基金项目
国家自然科学基金面上项目(81970664)
出版年
2024
国家科技期刊平台
NSTL
万方数据