抑制PCSK9对人主动脉内皮细胞钙化的保护作用  

The protective effect of inhibiting PCSK9 on calcification of human aortic endothelial cells

作　　者：靳天慧 陈亮[1,2] 刘叶红[1] 盛瑛 宗刚军 Jin Tianhui;Chen Liang;Liu Yehong(Dept 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)

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

出　　处：《安徽医科大学学报》2021年第8期1246-1250,共5页Acta Universitatis Medicinalis Anhui

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

摘　　要：目的探讨抑制前蛋白转化酶枯草溶菌素9(PCSK9)对地塞米松联合L-抗坏血酸和β甘油磷酸诱导的人主动脉内皮细胞(HAEC)钙化的影响。方法使用不同浓度(0、10、30、50 mmol/L)β甘油磷酸+50μg/ml L-抗坏血酸+100 nmol/L地塞米松刺激HAEC,筛选出诱导细胞钙化的最佳浓度,并验证HAEC钙化时,PCSK9的mRNA和蛋白表达。采用小干扰RNA(siRNA)敲低HAEC中PCSK9的表达,采用相同条件诱导细胞钙化,检测敲低PCSK9后HAEC中PCSK9及主要钙化指标的表达变化。茜素红S染色观察细胞内的钙化沉积。结果 (1)当β甘油磷酸浓度为30 mmol/L时,HAEC中主要钙化指标骨形态发生蛋白(BMP) 2、BMP4和Runt相关转录因子2(RUNX2)的mR NA和蛋白表达水平达到峰值,诱导钙化效果最佳;(2)诱导HAEC钙化后细胞内PCSK9的mR NA和蛋白表达水平明显升高;(3)敲低细胞内PCSK9的表达能够抑制体外诱导的HAEC钙化,钙化相关基因和蛋白的表达降低,细胞内钙盐沉积明显减少。结论 PCSK9的表达与HAEC钙化水平相关,敲低PCSK9可抑制地塞米松联合抗坏血酸和β甘油磷酸诱导的HAEC钙化。Objective To investigate the effect of inhibiting proprotein convertase subtilisin/kexin type 9(PCSK9)on the calcification of human aortic endothelial cells(HAEC)induced by dexamethasone combined with L-ascorbic acid andβ-glycerophosphate.Methods Different concentrations(0,10,30,50 mmol/L)βglycerophosphate+50μg/ml L-ascorbic acid+100 nmol/L dexamethasone were used to stimulate HAEC,and the optimal concentration was selected to induce cellular calcification.The mRNA and protein expression of PCSK9 in HAEC was verified.Small interfering RNA(siRNA)was used to knock down the expression of PCSK9 in HAEC,and cell calcification was induced by the same conditions,and the expression changes of PCSK9 and major calcification indexes in HAEC were detected after PCSK9 knockdown.Alizarin red S staining was used to observe the calcification deposition in the cells.Results①When theβglycerophosphate concentration was 30 mmol/L,the mRNA and protein expression levels of the main calcification indicators bone morphogenetic protein(BMP)2,BMP4 and Runt-related transcription factor 2(RUNX2)in HAEC reached the peak,and the effect of inducing calcification was the best;②The mRNA and protein expression levels of PCSK9 significantly increased after HAEC calcification was induced;③Knockdown of PCSK9 expression in cells could significantly inhibit HAEC calcification induced in vitro,the expression of calcification-related genes and proteins significantly reduced,and the intracellular calcium salt deposition significantly reduced.Conclusion The expression of PCSK9 is related to the calcification level of HAEC.Knockdown of PCSK9 can inhibit the calcification of HAEC induced by dexamethasone combined with ascorbic acid andβ-glycerophosphate.

关 键 词：前蛋白转化酶枯草溶菌素9 血管钙化 血管内皮细胞 

分 类 号：R543[医药卫生—心血管疾病]