[1]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(03):31-36.[doi:10.3969/j.issn.1671-7414.2025.03.006]
　LEI Ru,CAO Yong,ZHU Ming.Experimental Study on the Protective Mechanism of USP22/KDM3A Pathway Against Ischemia-reperfusion Injury in Sevoflurane Postconditioning Ischemic Hypoxic Cardiomyocyte Model[J].Journal of Modern Laboratory Medicine,2025,40(03):31-36.[doi:10.3969/j.issn.1671-7414.2025.03.006]
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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()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年03期
页码:
31-36
栏目:
论著
出版日期:
2025-05-15

文章信息/Info

Title:
Experimental Study on the Protective Mechanism of USP22/KDM3A Pathway Against Ischemia-reperfusion Injury in Sevoflurane Postconditioning Ischemic Hypoxic Cardiomyocyte Model
文章编号:
1671-7414（2025）03-031-06
作者:
雷 汝，曹 勇，朱 明
（眉山心脑血管病医院麻醉科，四川眉山 620010）
Author(s):
LEI Ru, CAO Yong, ZHU Ming
(Department of Anesthesiology, Meishan Cardiovascular and Cerebrovascular Disease Hospital, Sichuan Meishan 620010，China)
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
分类号:
R-332
DOI:
10.3969/j.issn.1671-7414.2025.03.006
文献标志码:
A
摘要:
目的探讨泛素特异性蛋白酶22（USP22）/赖氨酸特异性去甲基化酶3A（KDM3A）通路在七氟烷后处理（SPC）改善缺血缺氧心肌细胞再灌注损伤中的作用。方法将H9c2大鼠心肌细胞分为对照组、缺氧/复氧（H/R）组、H/R+七氟烷（Sevo）组、H/R+Sevo+si-NC组、H/R+Sevo+si-USP22组和H/R+Sevo+si-KDM3A组。采用qRT-PCR和Westernblot检测USP22和KDM3A的mRNA和蛋白表达；细胞计数试剂盒检测细胞活性；流式细胞术检测细胞凋亡率；ELISA检测肌酸激酶同工酶（CK-MB）、心肌肌钙蛋白I（cTnI）含量和乳酸脱氢酶（LDH）水平；免疫共沉淀实验检测USP22和KDM3A蛋白互作以及KDM3A泛素化。结果与对照组相比，H/R组细胞USP22mRNA和蛋白表达降低（t=49.574，14.852），细胞活性降低（t=46.597），细胞凋亡率，CK-MB，cTnI和LDH水平升高（t=17.722，21.346，22.863，9.722），KDM3A蛋白表达降低（t=15.879），泛素化水平升高，差异具有统计学意义（均P<0.01）；与H/R组比较，H/R+Sevo组H9c2细胞USP22的mRNA和蛋白表达升高（t=24.648，17.644），细胞活性升高（t=44.703），凋亡率，CK-MB，cTnI和LDH水平降低（t=13.736，18.018，17.012，13.856），KDM3A蛋白表达升高（t=12.970），泛素化水平降低，差异具有统计学意义（均P<0.01）。与H/R+Sevo+si-NC组相比，H/R+Sevo+si-USP22组细胞活力降低（t=20.785），凋亡率、CK-MB，cTnI和LDH水平升高（t=6.821，6.862，6.442，3.781），KDM3A蛋白表达降低（t=4.648），泛素化水平升高，差异具有统计学意义（均P<0.05）；USP22在KDM3A的启动子中富集，二者之间存在直接调控关系。与H/R+Sevo+si-NC组相比，H/R+Sevo+si-KDM3A组细胞活性降低（t=16.501），细胞凋亡率以及CK-MB，cTnI和LDH水平升高（t=8.954，10.533，6.801，8.004），差异具有统计学意义（均P<0.01）。结论SPC可减轻H/R诱导的H9c2心肌细胞损伤，其机制可能与上调USP22稳定KDM3A蛋白水平有关。
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

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备注/Memo

备注/Memo:
基金项目：四川省卫生健康科研课题普及项目（19PJ214）。
作者简介：雷汝（1983-），男，本科，主治医师，研究方向：麻醉学，E-mail：leiru1983@163.com。
更新日期/Last Update: 2025-05-15