成纤维细胞激活蛋白在小鼠心脏组织中时程表达变化及内肽酶活性测定  

Expression Profile and Endopeptidase Activity of Fibroblast Activation Protein in Mouse Heart Tissue

作　　者：廉虹[1] 刘维静 聂宇[1] LIAN Hong;LIU Wei-jing;NIE Yu(State Key Laboratory of Cardiovascular Diseases,Fuwai Hospital,National Center for Cardiovascular Disease,Chinese Academy of MedicalSciences&Peking Union Medical College,Beijing 100037;School of Basic Medical Sciences,Shanxi Medical University,Taiyuan 030001)

机构地区：[1]中国医学科学院北京协和医学院国家心血管病中心阜外医院心血管疾病国家重点实验室,北京市100037 [2]山西医科大学基础医学院,山西省太原市030001

出　　处：《中国分子心脏病学杂志》2020年第3期3408-3411,共4页Molecular Cardiology of China

基　　金：北京市自然科学基金(7182140);国家自然科学基金(81900343)。

摘　　要：目的了解小鼠出生后成纤维细胞激活蛋白(Fap)蛋白在心脏组织中的表达情况以及对其内肽酶活性进行测定。方法将1天、4天、7天、14天、21天、2月龄C57小鼠心肌组织进行取材,提取RNA和蛋白质,通过qPCR和Western blot方法检测Fap表达;分别通过RIPA裂解法、PBS研磨法和膜蛋白提取试剂盒提取小鼠心脏组织蛋白,测定Fap蛋白内肽酶活性曲线。结果生理情况下Fap在出生后的发育过程中蛋白表达逐渐升高,离乳后到成年期达到稳定状态;通过三种提取蛋白方法得到的酶活力曲线,确定三种蛋白提取方法对Fap蛋白内肽酶活性的测定无明显差异。结论在小鼠出生后Fap蛋白在心肌组织中表达量不断升高并在离乳后保持稳定;Fap内肽酶活性不受蛋白提取方法的影响,提示Fap的内肽酶活性中心很稳定。Objective To investigate the expression profile of fibroblast activation protein(Fap)in heart tissue of mouse after birth and to determine its endopeptidase activity.Methods RNA and protein were extracted from the heart tissues of C57 mice at different time points of 1 day,4 days,7 days,14 days,21 days,and 2 months after birth.Fap expression was determined by qPCR and Western blot.Heart tissues protein was extracted from heart tissue by RIPA lysing method,PBS grinding method or membrane protein extraction kit.Then the endopeptidase activity curves of Fap were measured.Results Under physiological conditions,Fap expression gradually increased and reached a steady state after weaning.Based on the enzyme activity curves obtained by the three protein extraction methods,there was no significant difference in endopeptidase activity of Fap.Conclusion The protein expression level of Fap in heart tissues increases after birth and remains stable after weaning.The endopeptidase activity of Fap is not affected by protein extraction methods,suggesting that the endopeptidase active center of Fap is stable.

关 键 词：成纤维细胞激活蛋白 蛋白提取方法 内肽酶活性 

分 类 号：R363[医药卫生—病理学]