现代检验医学杂志2025，Vol.40Issue(3)：31-36,6.DOI:10.3969/j.issn.1671-7414.2025.03.006
USP22/KDM3A通路对七氟烷后处理缺血缺氧心肌细胞模型再灌注损伤保护机制的实验研究
Experimental Study on the Protective Mechanism of USP22/KDM3A Pathway Against Ischemia-reperfusion Injury in Sevoflurane Postconditioning Ischemic Hypoxic Cardiomyocyte Model
摘要
Abstract
Objective To investigate the role of the ubiquitin specific protease 22(USP22)/lysine specific demethylase 3A(KDM3A)pathway in improving ischemia-reperfusion injury of hypoxic cardiomyocytes in sevoflurane postconditioning(SPC).Methods H9c2 rat cardiomyocytes were divided into control group,hypoxia/reoxygenation(H/R)group,H/R+sevoflurane(Sevo)group,H/R+Sevo+si-NC group,H/R+Sevo+si-USP22 group and H/R+Sevo+si-KDM3A group.Real time fluorescence quantitative PCR(qRT-PCR)and Western blotting were used to detect the mRNA and protein expression of USP22 and KDM3A,cell activity was detected by cell counting kit-8,apoptosis rate was detected by flow cytometry,and ELISA was used to detect the levels of creatine kinase MB(CK-MB)and cardiac troponin I(cTn I),while the assay kit was used to detect the levels of lactate dehydrogenase(LDH).Immunoprecipitation assay was used to detect the interaction between USP22 and KDM3A proteins,as well as KDM3A ubiquitination.Results Compared with the control group,the mRNA and protein expression of USP22 in H/R group cells decreased(t=49.574,14.852),cell activity decreased(t=46.597),cell apoptosis rate,CK-MB,cTn I and LDH levels increased(t=17.722,21.346,22.863,9.722),KDM3A protein expression decreased(t=15.879),and ubiquitination level increased,the differences were statistically significant(all P<0.01).Compared with the H/R group,the mRNA and protein expression of USP22 in H9c2 cells in the H/R+Sevo group increased(t=24.648,17.644),cell activity increased(t=44.703),apoptosis rate,CK-MB,cTn I and LDH levels decreased(t=13.736,18.018,17.012,13.856),KDM3A protein expression increased(t=12.970),ubiquitination level decreased,and the differences were statistically significant(all P<0.01).Compared with the H/R+Sevo+si-NC group,the cell viability of the H/R+Sevo+si-USP22 group was decreased(t=20.785),the apoptosis rate,CK-MB,cTn I and LDH levels were increased(t=6.821,6.862,6.442,3.781),the KDM3A protein expression was decreased(t=4.648),and the ubiquitination level was increased,and the differences were statistically significant(all P<0.05).USP22 was enriched in the promoter of KDM3A,and there was a direct regulatory relationship between USP22 and KDM3A.Compared with the H/R+Sevo+si-NC group,the cell viability was decreased(t=16.501),the apoptosis rate and the levels of CK-MB,cTn I and LDH were increased in the H/R+Sevo+si-KDM3A group(t=8.954,10.533 6.801,8.004),and the differences were statistically significant(all P<0.01).Conclusion SPC attenuates H/R-induced H9c2 cardiomyocyte injury,and the mechanism may be related to the up-regulation of USP22 stabilizing KDM3A protein levels.关键词
七氟烷后处理/泛素特异性蛋白酶22/赖氨酸特异性去甲基化酶3A/心肌缺血再灌注损伤Key words
sevoflurane postconditioning/ubiquitin specific protease 22/lysine specific demethylase 3A/myocardial ischemia-reperfusion injury引用本文复制引用
雷汝,曹勇,朱明..USP22/KDM3A通路对七氟烷后处理缺血缺氧心肌细胞模型再灌注损伤保护机制的实验研究[J].现代检验医学杂志,2025,40(3):31-36,6.基金项目
四川省卫生健康科研课题普及项目(19PJ214). （19PJ214）
