[1]陈梦雅 谢赛阳 邓伟.抑制泛素特异性蛋白酶7改善血管紧张素Ⅱ诱导的心肌细胞肥大[J].心血管病学进展,2024,(2):174.[doi:10.16806/j.cnki.issn.1004-3934.2024.02.016]
　CHEN Mengya,XIE Saiyang,DENG Wei.Inhibition of Ubiquitin-Specific Protease 7 Improves Angiotensin-Induced Cardiomyocyte Hypertrophy[J].Advances in Cardiovascular Diseases,2024,(2):174.[doi:10.16806/j.cnki.issn.1004-3934.2024.02.016]
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抑制泛素特异性蛋白酶7改善血管紧张素Ⅱ诱导的心肌细胞肥大()
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《心血管病学进展》[ISSN:51-1187/R/CN:1004-3934]

卷:
期数:
2024年2期
页码:
174
栏目:
论著
出版日期:
2024-02-25

文章信息/Info

Title:
Inhibition of Ubiquitin-Specific Protease 7 Improves Angiotensin-Induced Cardiomyocyte Hypertrophy
作者:
陈梦雅 谢赛阳 邓伟
(武汉大学人民医院心血管内科 代谢与相关慢病湖北省重点实验室,湖北 武汉 430060 )
Author(s):
CHEN Mengya; XIE Saiyang; DENG Wei
(Department of Cardiology,Renmin Hospital of Wuhan University,Hubei Key Laboratory of Metabolic and Chronic Diseases,Wuhan 430060 ,Hubei,China)
关键词:
泛素特异性蛋白酶7; 血管紧张素Ⅱ; FT671; 心肌肥厚; 氧化应激
Keywords:
Ubiquitin-specific protease 7; Angiotensin; FT671; Hypertrophy; Oxidative stress
DOI:
10.16806/j.cnki.issn.1004-3934.2024.02.016
摘要:
目的 探究泛素特异性蛋白酶7（USP7）抑制剂FT671在血管紧张素Ⅱ（AngⅡ）诱导的H9C2细胞肥大中的作用和潜在机制。方法 将H9C2细胞随机分为四组：对照组、FT671组、AngⅡ组、FT671+AngⅡ组。利用α-actinin细胞免疫荧光染色检测心肌细胞表面积；western blot检测心房钠尿肽（ANP）、脑钠肽（BNP），B细胞淋巴瘤2（BCL-2）、B细胞淋巴瘤2相关蛋白X（Bax）、白细胞介素-6（IL-6）、单核细胞趋化蛋白-1（MCP-1）、肿瘤坏死因子-α（TNF-α）、核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）以及NADPH氧化酶4（Nox4）的蛋白表达水平；RT-PCR检测 ANP 、BNP、肌球蛋白重链（β-MHC）、IL-6、MCP-1、TNF-α的mRNA表达；TUNEL染色检测细胞凋亡情况，细胞计数试剂盒8（CCK8）实验检测各组细胞活力，活性氧（ROS）染色检测细胞内氧化损伤水平。结果 与对照组相比，AngⅡ组USP7蛋白表达明显增加，而使用FT671后，USP7表达明显被抑制。与AngⅡ组相比，FT671+AngⅡ组心肌细胞面积减小；ANP、BNP、Bax的蛋白表达减少，ANP、BNP、β-MHC、Bax、IL-6、MCP-1以及TNF-α的mRNA表达减少；BCL-2的蛋白和mRNA表达均增加。与AngⅡ组相比，FT671+AngⅡ组TUNEL阳性细胞明显减少，心肌细胞活力增强， ROS 生成减少，Nox4、NLRP3蛋白表达减少，差异具有统计学意义（P<0.05）。结论 FT671通过抑制Nox4/ROS/NLRP3炎性通路减轻AngⅡ诱导的心肌细胞中的氧化应激和炎性反应，从而减轻心肌细胞肥大。
Abstract:
Objective To investigate the effect and mechanism of ubiquitin-specific protease 7(USP7) inhibitor FT671 in cardiomyocyte hypertrophy induced by angiotensinⅡ(AngⅡ). Methods H9C2 cardiomyocytes were randomly divided into four groups: control group,FT671 group,AngⅡ group ,FT671+AngⅡ group. The surface area of cardiomyocytes was detected by immunofluorescence staining of α-actinin. The protein expression of atrial natriuretic peptide (ANP),brain natriuretic peptide (BNP) ,B-cell lymphoma-2 (BCL-2),B-cell lymphoma-2-associated X protein (Bax),interleukin-6 (IL-6),monocyte chemoattractant protein-1 (MCP-1),tumor necrosis factor-α (TNF-α),nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and reduced nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4) were detected by western blot. The mRNA levels of ANP,BNP,β-myosin heavy chain (β-MHC),IL-6,MCP-1,TNF-α were detected by Real-Time PCR. TUNEL staining was used to evaluate cell apoptosis. Cell Counting Kit-8（CCK8） was used to detect cell viability. The fluorescent probe 2’,7’-dichlorofluorescein diacetate (DCFH-DA) was used to measure intracellular reactive oxygen species (ROS) formation. Results There was a significant increase in USP7 protein expression in the AngⅡ group,which was evidently inhibited by the USP7 inhibitor FT671. Compared with AngⅡ group,cardiomyocyte?surface area was significantly?reduced in FT671+AngⅡ group; the protein expression of ANP,BNP and Bax,as well as mRNA level of ANP,BNP,β-MHC,Bax,IL-6,MCP-1,TNF-α,were decreased in FT671+AngⅡ group. In addition,the protein expression and mRNA level of BCL-2 were increased in FT671+AngⅡ group. Additionally,compared with AngⅡ group,TUNEL positive cells was significantly reduced,cell viability was improved and ROS generation was inhibited in FT671+AngⅡ group. Further studies showed the protein expression of NLRP3 and Nox4 was decreased after FT671 treatment in AngⅡ induced cardiomyocyte hypertrophy. Conclusion The USP7 inhibitor FT671 attenuated oxidative stress and inflammatory response in AngⅡ-induced cardiomyocyte by inhibiting the Nox4/ROS/NLRP3 inflammatory pathway,thereby reducing cardiomyocyte hypertrophy.

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更新日期/Last Update: 2024-03-29