抑制NSF活性可导致囊泡转运蛋白和细胞膜蛋白表达异常及细胞凋亡  

Abnormal Expression of Vesicular Transport Proteins and Cell Membrane Proteins and Enhanced Apoptosis after Inhibition of NSF in Human Pulmonary Arterial Endothelial Cells

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(Department of Cardiology,Fuwai Hospital,National Center for Cardiovascular Disease,Chinese Academy of Medical Sciences&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,阜外医院心内科,北京市100037

出　　处：《中国分子心脏病学杂志》2020年第4期3486-3492,共7页Molecular Cardiology of China

基　　金：国家自然科学基金(81370326)。

摘　　要：目的探讨在人肺动脉内皮细胞(human pulmonary arterial endothelial cell,HPAEC)中抑制NSF活性后,囊泡转运蛋白NSF、α-SNAP、syntaxin 4和SNAP23的表达变化与细胞膜蛋白BMRP2、小凹蛋白-1和e NOS的表达变化和凋亡的相关性。方法在培养的HPAEC中加入NEM抑制NSF活性,观察HPAEC的形态变化,分别采用实时定量PCR和Western blot检测相关蛋白的表达。结果抑制NSF后,HPAEC在8~12 h开始出现胞体肿胀、变大等形态变化;NSF、α-SNAP、syntaxin 4和SNAP23的m RNA表达水平均显著增加,NSF和α-SNAP蛋白表达水平呈现轻度降低—增加—显著减少的趋势,而syntaxin 4和SNAP23蛋白均呈显著减少的趋势;BMPR2、小凹蛋白-1和e NOS的表达水平基本呈先增加后下降的趋势;凋亡蛋白caspase 8和fas在24 h后明显增加。结论抑制NSF活性可导致囊泡转运蛋白和细胞膜蛋白表达变化以及细胞凋亡。Objective To evaluate the gene expression of vesicle transport proteins NSF,α-SNAP,syntaxin 4 and SNAP23 in human pulmonary arterial endothelial cell(HPAEC)together with the expression of membrane proteins bone morphogenic protein receptor 2(BMPR2),caveolin-1(CAV-1)and endothelial nitric oxide synthase(eNOS),and cellular apoptosis.Methods N-ethyl maleimide was used to inhibit NSF in HPAEC.The cells were harvested for analysis after 4,8,12,24,48,72 and 96 hours after treatment.The mRNA and protein expression were studied by real time PCR and Western blot.Results After treatment with N-ethyl maleimide,HPAECs became swelling and enlarged since 8-12 hours;The mRNA expressions of NSF,α-SNAP,syntaxin 4 and SNAP23 were significantly increased;The protein expressions of NSF andα-SNAP were decreased at first,then increased and decreased at last;Syntaxin 4 and SNAP23 protein expressions were reduced from 4 hours after treatment;While the mRNA and protein expressions of BMPR2,CAV-1 and eNOS trended to increase at first and then decrease;The expressions of apoptotic proteins caspase 8 and fas were greatly enhanced since 24 hours after treatment.Conclusions Inhibition of NSF can lead to abnormal expressions of vesicular transport proteins NSF,α-SNAP,syntaxin 4 and SNAP23 and cellular membrane proteins BMPR2,CAV-1 and eNOS and increase cellular apoptosis in HPAEC.

关 键 词：人肺动脉内皮细胞 囊泡转运 N-乙基马来酰亚胺敏感因子 

分 类 号：R544.1[医药卫生—心血管疾病]