FoxO4 negatively modulates USP10 transcription to aggravate the apoptosis and oxidative stress of hypoxia/reoxygenation-induced cardiomyocytes by regulating the Hippo/YAP pathway
FoxO4 negatively modulates USP10 transcription to aggravate the apoptosis and oxidative stress of hypoxia/reoxygenation-induced cardiomyocytes by regulating the Hippo/YAP pathway
复制标题
DOI：
10.1007/s10863-021-09910-7
复制
发表时间：
2021-07-12
影响因子：
3
通讯作者：
Yang, Yan
中科院分区：
文献类型：
--
作者：
Huang, Jingwen;Liu, Yu;Yang, Yan
急性心肌梗死（AMI）是全球范围内的主要死亡原因。本研究旨在探讨叉头盒O 4（FoxO 4）是否可以通过Hippo/雅普途径负性调节泛蛋白特异性肽酶10（USP 10）转录，从而加剧缺氧/复氧（H/R）诱导的心肌细胞凋亡和氧化应激。RT-q PCR检测H/R诱导后H9 C2细胞中USP 10和FoxO 4的mRNA表达，Western blot检测转染后H9 C2细胞中USP 10和FoxO 4的蛋白表达。CCK-8法检测H/R诱导后H9 C2细胞存活率，TUNEL法检测转染后H9 C2细胞凋亡率。用合适的试剂盒分析H/R诱导或转染后H9 C2细胞中乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和丙二醛（MDA）的表达，用活性氧测定试剂盒检测细胞内活性氧（ROS）水平。采用双荧光素酶报告基因试验和染色质免疫沉淀（ChIP）来确认USP 10和FoxO 4的组合。Western blot检测凋亡相关蛋白和Hippo/雅普通路相关蛋白的表达。因此，在H/R诱导的H9 C2细胞中，USP 10表达以时间依赖性方式降低。USP 10过表达增加了H/R诱导的H9 C2细胞的活力，抑制了细胞凋亡和氧化应激。此外，FoxO 4调节USP 10转录。H/R诱导H9 C2细胞FoxO 4表达增加。FoxO 4过表达可通过调节Hippo/雅普信号通路逆转USP 10过表达对H/R诱导的H9 C2细胞的保护作用。综上所述，FoxO 4通过阻断Hippo/雅普通路，负性调节USP 10的转录，加重H/R诱导的H9 C2细胞凋亡和氧化应激。
Acute myocardial infarction (AMI) is the main cause of death in the whole world. This study aimed to investigate whether forkhead box O4 (FoxO4) could negatively modulate ubiquitin specific peptidase 10 (USP10) transcription to aggravate the apoptosis and oxidative stress of hypoxia/reoxygenation (H/R)-induced cardiomyocytes through Hippo/YAP pathway. mRNA expression as well as protein expressions of USP10 and FoxO4 in H9C2 cells after H/R induction or transfection were respectively detected by Reverse transcription-quantitative (RT-q) PCR analysis and Western blot. The viability and apoptosis of H9C2 cells after H/R induction or transfection were respectively detected by CCK-8 and TUNEL assays. The expressions of lactate dehydrogenase (LDH), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in H9C2 cells after H/R induction or transfection were analyzed using appropriate kits and intracellular reactive oxygen species (ROS) levels were detected using a ROS Assay Kit. Dual luciferase reporter assay and Chromatin Immunoprecipitation (ChIP) have adopted to confirm the combination of USP10 and FoxO4. Western blot was also used to analyze the expression of apoptosis-related proteins and Hippo/YAP pathway-related proteins. As a result, USP10 expression was decreased in H/R-induced H9C2 cells in a time-dependent manner. USP10 overexpression increased the viability and suppressed the apoptosis and oxidative stress of H/R-induced H9C2 cells. In addition, FoxO4 modulated USP10 transcription. FoxO4 expression was increased in H9C2 cells induced by H/R. FoxO4 overexpression could reverse the protective effects of USP10 overexpression on H/R-induced H9C2 cells by regulating the Hippo/YAP signaling pathway. In conclusion, FoxO4 negatively modulated USP10 transcription to aggravate the apoptosis and oxidative stress of H/R-induced H9C2 cells via blocking Hippo/YAP pathway.