[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]
点击复制

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()
分享到：

《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.

参考文献/References:

[1] McDonagh TA,Metra M,Adamo M,et al.2021 ESC Guidelines for the diagnosis and treatment of acute and chronic heart failure[J].Eur Heart J,2021,42(36):3599-3726.
[2] Xie S,Chen M,Fang W,et al. Diminished arachidonate 5-lipoxygenase perturbs phase separation and transcriptional response of Runx2 to reverse pathological ventricular remodeling[J].EBioMedicine,2022,86:104359.
[3] Swatek KN,Komander D. Ubiquitin modifications[J].Cell Res,2016,26(4):399-422.
[4] Zheng J,Chen C,Guo C,et al.The pleiotropic ubiquitin-specific peptidase 16 and its many substrates[J].Cells,2023,12(6):886.
[5] Clague MJ,Urbé S,Komander D. Breaking the chains: deubiquitylating enzyme specificity begets function[J].Nat Rev Mol Cell Biol,2019,20(6):338-352.
[6] Hu Y,Ma Z,Chen Z,et al.USP47 promotes apoptosis in rat myocardial cells after ischemia/reperfusion injury via NF-kappaB activation[J].Biotechnol Appl Biochem,2021,68(4):841-848.
[7] Han X,Zhang YL,Fu TT,et al.Blockage of UCHL1 activity attenuates cardiac remodeling in spontaneously hypertensive rats[J].Hypertens Res,2020,43(10):1089-1098.
[8] Bi HL,Zhang YL,Yang J,et al. Inhibition of UCHL1 by LDN-57444 attenuates Ang II-Induced atrial fibrillation in mice[J].Hypertens Res,2020,43(3):168-177.
[9] Pozhidaeva A,Bezsonova I. USP7: structure,substrate specificity,and inhibition[J].DNA Repair (Amst),2019,76:30-39.
[10] Fountain MD,Oleson DS,Rech ME,et al. Pathogenic variants in USP7 cause a neurodevelopmental disorder with speech delays,altered behavior,and neurologic anomalies[J].Genet Med,2019,21(8):1797-1807.
[11] Zhang XW,Feng N,Liu YC,et al. Neuroinflammation inhibition by small-molecule targeting USP7 noncatalytic domain for neurodegenerative disease therapy[J].Sci Adv,2022,8(32):eabo0789.
[12] Galarreta A,Valledor P,Ubieto-Capella P,et al. USP7 limits CDK1 activity throughout the cell cycle[J].EMBO J,2021,40(11):e99692.
[13] Xue Q,Yang D,Zhang J,et al. USP7,negatively regulated by miR-409-5p,aggravates hypoxia-induced cardiomyocyte injury[J].APMIS,2021,129(3):152-162.
[14] Suetomi T,Willeford A,Brand CS,et al. Inflammation and NLRP3 inflammasome activation initiated in response to pressure overload by Ca2+/calmodulin-dependent protein kinase Ⅱ delta signaling in cardiomyocytes are essential for adverse cardiac remodeling[J].Circulation,2018,138(22):2530-2544.
[15] Bedard K,Krause KH. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology[J].Physiol Rev,2007,87(1):245-313.
[16] Nie L,Wang C,Liu X,et al. USP7 substrates identified by proteomics analysis reveal the specificity of USP7[J].Genes Dev,2022,36(17-18):1016-1030.
[17] Meulmeester E,Maurice MM,Boutell C,et al.Loss of HAUSP-mediated deubiquitination contributes to DNA damage-induced destabilization of Hdmx and Hdm2[J].Mol Cell,2005,18(5):565-276.
[18] Cummins JM,Rago C,Kohli M,et al.Tumour suppression: disruption of HAUSP gene stabilizes p53[J].Nature,2004,428(6982):1 p following 486.
[19] Song MS,Salmena L,Carracedo A,et al. The deubiquitinylation and localization of PTEN are regulated by a HAUSP-PML network[J].Nature,2008,455(7214):813-817.
[20] Liu G,Liu Q,Yan B,et al.USP7 inhibition alleviates H2O2-induced injury in chondrocytes via inhibiting NOX4/NLRP3 pathway[J].Front Pharmacol,2020,11:617270.
[21] Ji L,Lu B,Zamponi R,et al. USP7 inhibits Wnt/beta-catenin signaling through promoting stabilization of Axin[J].Nat Commun,2019,10(1):4184.
[22] Sanz RL,Inserra F,García Menéndez S,et al.Metabolic syndrome and cardiac remodeling due to mitochondrial oxidative stress involving gliflozins and sirtuins[J].Curr Hypertens Rep,2023;25(6):91-106.
[23] Daou D,Gillette TG,Hill JA. Inflammatory mechanisms in heart failure with preserved ejection fraction[J].Physiology (Bethesda),2023,38(5):0.
[24] Hu C,Zhang X,Wei W,et al. Matrine attenuates oxidative stress and cardiomyocyte apoptosis in doxorubicin-induced cardiotoxicity via maintaining AMPKalpha/UCP2 pathway[J].Acta Pharm Sin B,2019,9(4):690-701.
[25] Werbner B,Tavakoli-Rouzbehani OM,Fatahian AN,et al.The dynamic interplay between cardiac mitochondrial health and myocardial structural remodeling in metabolic heart disease,aging,and heart failure[J].J Cardiovasc Aging,2023,3(1):9.
[26] Guo S,Chen X. The human Nox4:gene,structure,physiological function and pathological significance[J].J Drug Target,2015,23(10):888-896.
[27] Xu Q,Liu M,Gu J,et al. Ubiquitin-specific protease 7 regulates myocardial ischemia/reperfusion injury by stabilizing Keap1[J].Cell Death Discov,2022,8(1):291.
[28] Palazón-Riquelme P,Worboys JD,Green J,et al. USP7 and USP47 deubiquitinases regulate NLRP3 inflammasome activation[J].EMBO Rep,2018,19(10):e44766.

相似文献/References:

[1]王海星,尹德春,曲秀芬.病态窦房结综合征的心房纤维化血清学指标研究进展[J].心血管病学进展,2020,(3):310.[doi:10.16806/j.cnki. issn.1004-3934.2020.03.023]
　WANG Haixing,YIN Dechun,QU Xiufen.Serological Indicators of Atrial Fibrosis for Sick Sinus Syndrome[J].Advances in Cardiovascular Diseases,2020,(2):310.[doi:10.16806/j.cnki. issn.1004-3934.2020.03.023]
[2]王欣笛,苏冠华. MicroRNA-155在高血压发病机制和治疗中的研究进展[J].心血管病学进展,2020,(10):1040.[doi:10.16806/j.cnki.issn.1004-3934.2020.10.010]
　WANG Xindi,SU Guanhua.MicroRNA-155 in Pathogenesis and Treatment of Hypertension[J].Advances in Cardiovascular Diseases,2020,(2):1040.[doi:10.16806/j.cnki.issn.1004-3934.2020.10.010]
[3]王璐 桑婉玥 简易 韩亚凡 王菲菲 李耀东.抑制MAPK14通过减轻线粒体自噬改善AngⅡ诱导的心房颤动[J].心血管病学进展,2024,(4):373.[doi:10.16806/j.cnki.issn.1004-3934.2024.04.019]
　WANG Lu,SANG Wanyue,JIAN Yi,et al.Inhibition of MAPK14 Improves Ang-Induced Atrial Fibrillation by Reducing Mitochondrial Autophagy[J].Advances in Cardiovascular Diseases,2024,(2):373.[doi:10.16806/j.cnki.issn.1004-3934.2024.04.019]
[4]李岚岚 谢赛阳 邓伟.tirzepatide对AngⅡ诱导的心肌细胞肥大的抑制作用及机制研究[J].心血管病学进展,2024,(9):844.[doi:10.16806/j.cnki.issn.1004-3934.2024.09.017]
　LI Lanlan,XIE Saiyang,DENG Wei.Inhibitory Effect of Tirzepatide on Ang-induced Cardiomyocyte Hypertrophy and Study of the Mechanism[J].Advances in Cardiovascular Diseases,2024,(2):844.[doi:10.16806/j.cnki.issn.1004-3934.2024.09.017]

更新日期/Last Update: 2024-03-29