NSF和α-SNAP在吡咯野百合碱处理肺动脉内皮细胞中的表达  

Expression of Vesicular Transport Proteins NSF andα-SNAP in Human Pulmonary Arterial Endothelial Cell Treated by Monocrotaline Pyrrole

作　　者：张洪亮[1] 罗勤[1] 王勇[1] 赵智慧[1] 柳志红[1] ZHANG Hong-liang;LUO Qin;WANG Yong;ZHAO Zhi-hui;LIU Zhi-hong(Fuwai Hospital,National Center for Cardiovascular Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,China)

机构地区：[1]中国医学科学院北京协和医学院国家心脏病中心阜外医院,北京市100037

出　　处：《中国分子心脏病学杂志》2020年第6期3651-3656,共6页Molecular Cardiology of China

基　　金：国家自然科学基金(81641005)。

摘　　要：目的探讨囊泡转运蛋白NSF和α-SNAP在吡咯野百合碱(monocrotaline pyrrole,MCTP)处理人肺动脉内皮细胞(human pulmonary arterial endothelial cell,HPAEC)中的表达变化以及与膜蛋白小凹蛋白-1(caveolin-1,CAV-1)和内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)的表达变化和凋亡间的相关性。方法:应用MCTP处理培养的HPAEC。分别在处理后4、8、12、24、48和72小时通过实时定量PCR和Wester blot检测mRNA和蛋白的表达。结果MCTP处理后,HPAEC在12~24 h开始出现胞体肿胀、变大等形态变化;NSF和α-SNAP的mRNA和蛋白以及eNOS的mRNA表达从4~24 h开始呈先增加之后降低的规律;CAV-1 mRNA表达变化程度较小,CAV-1蛋白在12 h开始一致性下降;eNOS蛋白表达在大部分时间均下降;凋亡蛋白Bax表达在24 h后明显增加。结论MCTP所致的HPAEC损伤反应伴有囊泡转运蛋白NSF和α-SNAP的表达水平变化。Objective To evaluate the gene expression of vesicle transport proteins NSF andα-SNAP in human pulmonary arterial endothelial cell(HPAEC)treated by monocrotaline pyrrole(MCTP)together with the expression of membrane proteins caveolin-1(CAV-1)and endothelial nitric oxide synthase(eNOS),and apoptotic protein.Methods MCTP was used to treat HPAEC.The cells were harvested for analysis after 4,8,12,24,48 and 72 hours after treatment.The m RNA and protein expression were studied by real time PCR and Western blot respected.Results After treatment with MCTP,HPAECs became swelling and enlarged since 12-24 hours;The mRNA and protein expression of NSF andα-SNAP and mRNA level of eNOS started to increase from 4-24 hours and then decreased;The expression change of CAV-1 was rather little and the protein level of CAV-1 declined continuously from 12 hours;eNOS protein expression was decreased in most time;While the amount of apoptotic protein Bax was greatly enhanced since 24 hours.Conclusion The change in mRNA and protein expressions of vesicular transport proteins NSF andα-SNAP was involved in the pathology of MCTP induced damage to MCTP.

关 键 词：动脉型肺动脉高压 人肺动脉内皮细胞 N-乙基马来酰亚胺敏感因子 α-可溶性N-乙基马来酰亚胺敏感因子附着蛋白 

分 类 号：R544.1[医药卫生—心血管疾病]