Inhibition of Ubiquitin-Specific Protease 7 Improves AngiotensinⅡ-Induced Cardiomyocyte Hypertrophy
Objective To investigate the effect and mechanism of ubiquitin-specific protease 7(USP7)inhibitor FT671 in cardiomyocyte hypertrophy induced by angiotensin Ⅱ(Ang Ⅱ).Methods H9c2 cardiomyocytes were randomly divided into four groups:control group,FT671 group,Ang Ⅱ group,FT671+Ang Ⅱ 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),NOD-like receptor thermal protein domain associated protein 3(NLRP3)and 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 Ang Ⅱ group,which was evidently inhibited by the USP7 inhibitor FT671.Compared with Ang Ⅱ group,cardiomyocyte surface area was significantly reduced in FT671+Ang Ⅱ 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+AngⅡgroup.In addition,the protein expression and mRNA level of Bcl-2 were increased in FT671+Ang Ⅱ group.Additionally,compared with AngⅡ group,TUNEL positive cells was significantly reduced,cell viability was improved and ROS generation was inhibited in FT671+Ang Ⅱgroup.Further studies showed the protein expression of NLRP3 and Nox4 was decreased after FT671 treatment in Ang Ⅱ induced cardiomyocyte hypertrophy.Conclusion The USP7 inhibitor FT671 attenuated oxidative stress and inflammatory response in Ang Ⅱ-induced cardiomyocyte by inhibiting the Nox4/ROS/NLRP3 inflammatory pathway,thereby reducing cardiomyocyte hypertrophy.
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维普
