HRD1 介导的 HDAC2 泛素化调节 PPARα 介导的自噬,缓解代谢相关性脂肪肝。

IF 4.6 2区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Yina Wang , Yuanguo Chen , Xiao Xiao , Silei Deng , Jingjie Kuang , Yayong Li
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引用次数: 0

摘要

背景:代谢相关性脂肪肝(MAFLD)是全球慢性肝病的主要病因。自噬在脂质代谢中起着举足轻重的作用;然而,自噬活性在 MAFLD 中降低的机制仍不明确:方法:通过TUNEL检测和LC3免疫荧光染色监测自噬。Western印迹法检测自噬相关蛋白、PPARα、HDAC2和HRD1的表达。染色质免疫沉淀(ChIP)和双荧光素酶测定评估了HDAC2和PPARα启动子之间的关联,共免疫沉淀(co-IP)检测了HRD1介导的HDAC2泛素蛋白酶体降解。体外研究结果在缺氧诱导的 MAFLD 小鼠模型中得到了验证。通过苏木精和伊红染色、Masson三色染色和TUNEL检测法检测了肝组织的组织学变化、纤维化和细胞凋亡。通过 IHC 分析检测了关键分子的免疫活性:结果:缺氧抑制了肝细胞的自噬。低氧暴露下调了肝细胞中的 HRD1 和 PPARα,同时上调了 HDAC2。过量表达PPARα可促进肝脏自噬,而敲除HDAC2或过量表达HRD1可减少肝细胞中受缺氧抑制的自噬。从机理上讲,HDAC2是PPARα的转录抑制因子,而HRD1则通过泛素-蛋白酶体途径介导HDAC2的降解。功能研究进一步表明,缺氧通过HRD1/HDAC2/PPARα轴在体外和体内抑制肝脏自噬:结论:HRD1介导的HDAC2泛素化调节了PPARα介导的自噬,改善了缺氧诱导的MAFLD。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

HRD1-mediated ubiquitination of HDAC2 regulates PPARα-mediated autophagy and alleviates metabolic-associated fatty liver disease

HRD1-mediated ubiquitination of HDAC2 regulates PPARα-mediated autophagy and alleviates metabolic-associated fatty liver disease

Background

Metabolic-associated fatty liver disease (MAFLD) is a leading cause of chronic liver disease worldwide. Autophagy plays a pivotal role in lipid metabolism; however, the mechanism underlying the reduced autophagic activity in MAFLD remains elusive.

Methods

Autophagy was monitored by TUNEL assay and immunofluorescence staining of LC3. The expression of autophagy-related proteins, PPARα, HDAC2, and HRD1 was detected by Western blot. The association between HDAC2 and PPARα promoter was assessed by chromatin immunoprecipitation (ChIP) and dual-luciferase assays, and the HRD1-mediated ubiquitin-proteasomal degradation of HDAC2 was detected by co-immunoprecipitation (co-IP). The in vitro findings were validated in a hypoxia-induced MAFLD mouse model. Histological changes, fibrosis, and apoptosis in liver tissues were detected by hematoxylin and eosin staining, Masson's trichrome staining, and TUNEL assay. The immunoreactivities of key molecules were examined by IHC analysis.

Results

Hypoxia-suppressed autophagy in hepatocytes. Hypoxic exposure downregulated HRD1 and PPARα, while upregulating HDAC2 in hepatocytes. Overexpression of PPARα promoted hepatic autophagy, while knocking down HDAC2 or overexpressing HRD1 reduced hypoxia-suppressed autophagy in hepatocytes. Mechanistically, HDAC2 acted as a transcriptional repressor of PPARα, and HRD1 mediated the degradation of HDAC2 through the ubiquitin-proteasome pathway. Functional studies further showed that hypoxia-suppressed hepatic autophagy via the HRD1/HDAC2/PPARα axis in vitro and in vivo.

Conclusion

HRD1-mediated ubiquitination of HDAC2 regulates PPARα-mediated autophagy and ameliorates hypoxia-induced MAFLD.

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来源期刊
CiteScore
10.00
自引率
2.00%
发文量
151
审稿时长
44 days
期刊介绍: BBA Molecular Cell Research focuses on understanding the mechanisms of cellular processes at the molecular level. These include aspects of cellular signaling, signal transduction, cell cycle, apoptosis, intracellular trafficking, secretory and endocytic pathways, biogenesis of cell organelles, cytoskeletal structures, cellular interactions, cell/tissue differentiation and cellular enzymology. Also included are studies at the interface between Cell Biology and Biophysics which apply for example novel imaging methods for characterizing cellular processes.
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