抑制剂KN-93干预二乙酰吗啡致心肌细胞节律异常的蛋白质组学分析  

Proteomics analysis of diacetylmorphine-induced rhythm abnormalities in cardiac muscle cells interfered with by the inhibitor KN-93

作　　者：季敏 苏丽萍[2] 刘丽[2] 管雅玲 肖锦玲 蒲红伟[3] JI Min;SU Liping;LIU Li;GUAN Yalin;XIAO Jinling;PU Hongwei(College of Basic Medical Sciences,Xinjiang Medical University,Xinjiang Uygur Autonomous Region,Urumqi830011,China;Department of Pathology,the First Affiliated Hospital of Xinjiang Medical University,Xinjiang Uygur Autonomous Region,Urumqi830054,China;Department of Discipline Construction,the First Affiliated Hospital of Xinjiang Medical University,Xinjiang Uygur Autonomous Region,Urumqi830054,China)

机构地区：[1]新疆医科大学基础医学院,新疆乌鲁木齐830011 [2]新疆医科大学第一附属医院病理科,新疆乌鲁木齐830054 [3]新疆医科大学第一附属医院学科建设科,新疆乌鲁木齐830054

出　　处：《中国医药导报》2023年第19期4-9,共6页China Medical Herald

基　　金：国家自然科学基金资助项目(81860049、82160055)。

摘　　要：目的使用蛋白质组学分析抑制剂KN-93干预二乙酰吗啡致心肌细胞节律异常的作用。方法60只SPF级SD大鼠乳鼠,雌雄不限,提取心肌组织,体外培养SD大鼠乳鼠原代心肌细胞,分为对照组和实验组,对照组使用100μmol/L二乙酰吗啡作用24 h,实验组使用100μmol/L二乙酰吗啡+1μmol/L KN-93作用24 h。采用TMT相对定量蛋白质技术筛选出对照组和实验组之间具有显著性表达差异蛋白质,筛选标准:表达倍数上调>1.2倍或下调<0.83倍且P<0.05。通过GO功能富集分析差异表达蛋白质的主要功能、细胞定位和参与的生物学过程,通过KEGG通路富集分析差异表达蛋白质可能参与的代谢或信号通路,通过蛋白质相互作用网络分析蛋白质之间有结合作用或相互作用。结果与对照组比较,实验组Ca2+/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)磷酸化水平降低,差异有统计学意义(P<0.05);TMT技术共筛选出346个差异表达蛋白质,主要参与对细胞因子的反应、细胞对细胞因子刺激的反应等过程,影响相同的蛋白质结合、信号受体结合等分子功能和细胞外区域部分、胞外区等细胞组分蛋白,主要富集在癌症途径、PI3K-Akt信号通路、人乳头瘤病毒感染、阿尔茨海默病等信号通路。蛋白互作网络显示,MSH6、NFk B1等可能与CaMKⅡ存在蛋白质互相作用关系。与对照组比较,实验组ADCY7、NFk B1、MSH6、MMP2、MMP9、NQO1、TXNRD1蛋白表达显著降低,差异有统计学意义(P<0.05)。结论CaMKⅡ可能通过调控NFk B、MSH6等蛋白的改变,通过癌症途径信号通路调节心肌细胞凋亡,参与二乙酰吗啡致心肌细胞节律异常过程。Objective To analysis the role of the inhibitor KN-93 to intervene in diacetylmorphine-induced rhythm abnormalities in cardiac muscle cells.Methods Myocardial tissue was extracted from 60 SPF-grade SD rats,regardless of male or female.Primary myocardial cells of SD rats were cultured in vitro and divided into the control group and the experimental group.The control group was treated with 100μmol/L diacetylmorphine for 24 h,and the experimental group was treated with 100μmol/L diacetylmorphine+1μmol/L KN-93 for 24 h.The TMT relative quantitative protein technique was used to screen out the proteins with significant expression difference between the control group and the experimental group.The screening criteria were:up-regulation of expression ploidy>1.2-fold or down-regulation of expression ploidy<0.83-fold and P<0.05.The main functions,cellular localization,and biological processes involved in differentially expressed proteins were analyzed by GO functional enrichment.The possible metabolic or signaling pathways involved in differentially expressed proteins were analyzed by KEGG pathway enrichment.Protein interaction network was used to analyze the binding or interaction between proteins.Results Compared with the control group,the phosphorylation level of Ca2+/Calmodulin-dependent protein kinaseⅡ(CaMKⅡ)in the experimental group was decreased,and the difference was statistically significant(P<0.05).A total of 346 differentially expressed proteins were screened by TMT technology,which were mainly involved in the reaction to cytokines and cell response to cytokine stimulation and other processes,and affected molecular functions such as the same protein binding and signal receptor binding,as well as cellular component proteins such as extracellular regions and extracellular regions.It is mainly concentrated in cancer pathway,PI3K-Akt signaling pathway,human papillomavirus infection,Alzheimer disease,and other signaling pathways.The protein interaction network showed that MSH6,NFkB1,and CaMKⅡmight have prote

关 键 词：二乙酰吗啡 心肌细胞 Ca2+/钙调蛋白依赖性蛋白激酶Ⅱ 蛋白质组学 

分 类 号：R54[医药卫生—心血管疾病]