敲低前蛋白转化酶枯草溶菌素9(PCSK9)抑制人主动脉内皮细胞的内皮间质转化  

Knockdown of PCSK9 inhibits endothelial-to-mesenchymal transition of human aortic endothelial cells

作　　者：靳天慧 陈亮[1,2] 刘叶红[1,2] 盛瑛 周雨婷 宣诗怡 宗刚军 JIN Tianhui;CHEN Liang;LIU Yehong;SHENG Ying;ZHOU Yuting;XUAN Shiyi;ZONG Gangjun(Department of Cardiology,Wuxi Clinical College,Anhui Medical University,Wuxi 214044;Dept of Cardiology,No.904 Hospital of the PLA Joint Logistic Support Force,Wuxi 214044,China)

机构地区：[1]安徽医科大学无锡临床学院心内科,江苏无锡214044 [2]中国人民解放军联勤保障部队第904医院心内科,江苏无锡214044

出　　处：《细胞与分子免疫学杂志》2021年第12期1079-1084,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省青年医学重点人才基金(QNRC2016883);无锡市医疗与公众健康技术创新应用项目(N20192007)。

摘　　要：目的以人主动脉内皮细胞(HAEC)为研究对象,探讨敲减前蛋白转化酶枯草溶菌素9(PCSK9)对β甘油磷酸、地塞米松、L-抗坏血酸诱导内皮间质转化(EndoMT)的保护作用。方法以(0、10、30、50)mmol/Lβ甘油磷酸联合100 nmol/L地塞米松和50μg/ml L-抗坏血酸诱导HAEC建立EndoMT模型。采用PCSK9的特异性小干扰RNA(si-PCSK9)转染HAEC,实时荧光定量PCR与Western blot法检测HAEC的PCSK9的mRNA和蛋白表达。将HAEC随机分为空白组、EndoMT组、转染阴性对照小干扰RNA的EndoMT组、转染si-PCSK9的EndoMT组。实时荧光定量PCR检测α平滑肌肌动蛋白(α-SMA)、成纤维细胞特异蛋白1(FSP1)、血管内皮钙黏蛋白(VE-cadherin)的mRNA水平,Western blot法检测α-SMA、VE-cadherin的蛋白水平,免疫荧光染色法检测α-SMA的表达。结果30 mmol/Lβ甘油磷酸诱导EndoMT效果最佳,发生EndoMT时,PCSK9的mRNA及蛋白表达上调。而PCSK9敲减后,α-SMA、FSP1的表达下调,VE-cadherin的表达上调。结论敲低PCSK9可抑制HAEC的EndoMT。Objective To investigate the protective effect of proprotein convertase subtilisin/kexin type 9(PCSK9)knockdown on endothelial-to-mesenchymal transition(EndoMT)induced byβglycerophosphate,dexamethasone,and L-ascorbic acid in human aortic endothelial cells(HAECs).Methods EndoMT model was established by inducing HAECs with(0,10,30,50)mmol/L ofβglycerophosphate combined with 100 nmol/L of dexamethasone and 50μg/mL of L-ascorbic acid.HAECs were transfected with specific small interfering RNA of PCSK9(si-PCSK9),and the mRNA and protein expression levels of PCSK9 in HAECs were detected by real-time quantitative PCR and Western blotting.HAECs were randomized into blank group,EndoMT group,EndoMT group transfected with negative control small interfering RNA(si-NC),and EndoMT group transfected with si-PCSK9.The mRNA levels ofα-smooth muscle actin(α-SMA),fibroblast-specific protein 1(FSP1),and vascular endothelial cadherin(VE-cadherin)were detected by real-time quantitative PCR,the protein levels ofα-SMA and VE-cadherin were detected by Western blotting,and the expression ofα-SMA was detected by immunofluorescence staining.Results 30 mmol/L ofβglycerophosphate had the best effect in inducing EndoMT,and the expression of PCSK9 mRNA and protein was up-regulated when EndoMT occurred.After PCSK9 knockdown,the expressions ofα-SMA and FSP1 were down-regulated,while the expression of VE cadherin was up-regulated.Conclusion Knockdown of PCSK9 inhibits the EndoMT of HAECs.

关 键 词：内皮间质转化(EndoMT) 前蛋白转化酶枯草溶菌素9(PCSK9) 人主动脉内皮细胞(HAEC) 

分 类 号：R392-33[医药卫生—免疫学] R543[医药卫生—基础医学]