HDAC6抑制剂通过保护肾小球内皮细胞线粒体改善糖尿病肾病
DOI:
作者:
作者单位:

井冈山大学附属医院

作者简介:

通讯作者:

中图分类号:

基金项目:

江西省青年科学基金资助项目 (20192BAB215051)


HDAC6 inhibitors improve diabetic nephropathy by protecting glomerular endothelial cell mitochondria
Author:
Affiliation:

Affiliated Hospital of Jinggangshan University

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的 探讨组蛋白去乙酰化酶6(HDAC6)特异性小分子抑制剂Tubastatin A对糖尿病肾病(DN)小鼠肾损伤的保护作用和机制。方法 C57BL /6小鼠随机分为3组:对照组、DN组和Tubastatin A组。DN组和Tubastatin A组行包膜下肾切除术以切除右肾,并通过腹膜内注射STZ诱导DN。Tubastatin A组接受Tubastatin A治疗,每3天一次,连续治疗8周。RNA测序分析DN组和Tubastatin A组肾脏组织中差异表达基因。通过透射电子显微镜评估线粒体受损情况,以及DHE染色估计肾组织中的ROS水平。将小鼠肾小球内皮细胞(mGEC)暴露于高葡萄糖培养基(HG))或40 mM甘露醇(对照),加入或不加入Tubastatin A处理。采用蛋白质印迹分析HDAC6、肾损伤标志物KIM1和EMT标志物的表达情况,以及流式细胞仪检测细胞中线粒体ROS和细胞凋亡情况。结果 DN小鼠肾组织和暴露于HG的mGEC细胞中HDAC6表达上调,并与物KIM1水平升高一致。组织学分析显示DN小鼠的显著形态学变化,包括肾小球肥大、肾小球系膜基质积聚、肾小球基底膜增厚、肾小管基底膜增厚和出现肾小球、肾小管间质纤维化;Tubastatin A治疗缓解了这些变化。与对照DMSO相比,Tubastatin A在HG处理下显著降低了mGEC细胞中KIM1、HDAC6、α-SMA、N-钙粘蛋白、波形蛋白表达(P<0.05),并上调E-钙粘蛋白表达(P<0.05)。RNA测序揭示了揭示了与DN组小鼠相比,Tubastatin A组小鼠肾脏组织中与ECM-受体相互作用和与三羧酸(TCA)循环相关的基因富集。透射电子显微镜显示Tubastatin A组小鼠的肾小球内皮细胞受损线粒体的比例较DN组显著降低(P<0.01)。DHE染色显示Tubastatin A组小鼠肾组织中ROS水平较DN组降低(P<0.01)。在mGEC细胞中,Tubastatin A处理下调了HG诱导的mGEC细胞中的线粒体ROS水平(P<0.01),以及减少了细胞凋亡(P<0.05)。结论 Tubastatin A改善了HG诱导的肾小球内皮细胞损伤和DN进展,其作用机制与保护线粒体稳态和抑制EMT发生相关。

    Abstract:

    objective: To investigate the protective effect and mechanism of histone deacetylase 6 (HDAC6) specific small molecule inhibitor Tubastatin A on renal injury in diabetic nephropathy (DN) mice. Methods: C57BL/6 mice were randomly divided into 3 groups: control group, DN group and Tubastatin A group. Mice in the DN group and Tubastatin A group were intraperitoneally injected with燬TZ?80爉g/kg) daily for 3 days after the removal of one kidney. Tubastatin A group received Tubastatin A treatment every 3 days for 8 weeks. RNA sequencing analysis of differentially expressed genes in kidney tissue of DN group and Tubastatin A group. Mitochondrial damage was assessed by transmission electron microscopy, and ROS levels in kidney tissue were estimated by DHE staining. Mouse glomerular endothelial cells (mGEC) were exposed to high glucose medium (HG) or 40 mM mannitol (control) with or without Tubastatin A treatment. Western blot analysis was used to analyze the expression of HDAC6, kidney injury markers KIM1 and EMT markers, and flow cytometry was used to detect mitochondrial ROS and apoptosis in cells. Results: HDAC6 expression was up-regulated in DN mouse kidney tissue and mGEC cells exposed to HG, consistent with increased levels of KIM1. Histological analysis showed significant morphological changes in DN mice, including glomerular hypertrophy, mesangial matrix accumulation, glomerular basement membrane thickening, tubular basement membrane thickening and the presence of glomerular, intertubular fibrosis; Tubastatin A treatment alleviated these changes. Compared with control DMSO, Tubastatin A significantly decreased the expression of KIM1, HDAC6, α-SMA, N-cadherin, vimentin (P<0.05) and up-regulated the expression of E-cadherin (P<0.05) in mGEC cells under HG treatment. RNA-sequencing revealed the enrichment of genes related to ECM-receptor interaction and tricarboxylic acid (TCA) cycle in the kidney tissue of Tubastatin A mice compared with DN mice. Transmission electron microscopy showed that the proportion of damaged mitochondria in glomerular endothelial cells in Tubastatin A group was significantly lower than that in DN group (P<0.01). DHE staining showed that the level of ROS in the kidney tissue of Tubastatin A group was lower than that of DN group (P<0.01). In mGEC cells, Tubastatin A treatment down-regulated HG-induced mitochondrial ROS levels in mGEC cells (P<0.01), and reduced apoptosis (P<0.05). Conclusion: Tubastatin A ameliorates HG-induced glomerular endothelial cell injury and DN progression, and its mechanism is related to the protection of mitochondrial homeostasis and inhibition of EMT.

    参考文献
    相似文献
    引证文献
引用本文
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2022-09-23
  • 最后修改日期:2023-04-13
  • 录用日期:2023-10-19
  • 在线发布日期:
  • 出版日期:
自2024年1期开始,杂志参考文献改为中英文对照,具体格式要求可置下载中心查看!
关闭