摘要
目的:探讨长链非编码核糖核酸H19(IncRNA)对人主动脉平滑肌细胞(HA-VSMCs)自噬及表型转化的调控作用。方法:药物诱导HA-VSMCs发生表型转化,检测细胞内成骨相关蛋白RUNX2及平滑肌标志物α-SMA的表达水平,通过茜素红s染色及碱性磷酸酶ALP含量测定实验检测钙化水平,通过检测泛素化连接蛋白LC-3、P62观察自噬水平变化。结果:与正常组相比,钙化诱导HA-VSMCs组lncRNA H19表达量明显升高,成骨相关蛋白RUNX2表达水平增加,平滑肌标志物α-SMA表达下调,自噬被抑制。敲低lncRNA H19后,平滑肌细胞表型转化减缓,钙化程度减弱,自噬水平有所回升。结论:lncRNA H19可促进HA-VSMCs的表型转化及动脉钙化的发生,其机制可能与抑制HA-VSMCs的保护性自噬有关。
Objective:To investigate the effect of lncRNA H19 on phenotype transformation and autophagy in human aortal vascular smooth muscle cells(HA-VSMCs).Methods:HA-VSMCs were induced to transform into osteogenic phenotype up to 14 days.We detected the expression level of runt-related transcription factor 2(RUNX2),andα-smooth muscle actin(α-SMA)to evaluate the phenotype transformation of HAVSMCs.Alizarin Red S staining and the concentration of alkaline phosphatase(ALP)in HA-VSMCs were used to observe the extent of atrial calcification.The level of autophagy was reflected by ubiquitin-like modifier protein LC-3,P62.Results:Compared to the normal group,calcific group showed significantly high expression level of lncRNA H19,so were RUNX2.We also found the inhibition of LC-3 in calcific group and P62 showed the opposite trend.However,when we knocked down lncRNA H19,the expression level of RUNX2 were less than calcific group andα-SMA showed a re-rising trend.Comparing with the calcific group,Alizarin Red S staining and ALP experiment also indicated the extent of calcification was lower in si-H19 group.The inhibition of autophagy was removed in si-H19 group.Conclusions:lncRNA H19 promotes the phenotype transformation of HAVSMCs which leads to atrial calcification,and its potential mechanism is related to the inhibition of autophagy.
作者
杨佳奇
杨杰
孙铁男
周玉杰
YANG Jiaqi;YANG Jie;SUN Tienan;ZHOU Yujie(Department of Cardiology,Beijing Anzhen Hospital,Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Diseases,Beijing 100029,China)
出处
《心肺血管病杂志》
2019年第10期1060-1065,共6页
Journal of Cardiovascular and Pulmonary Diseases
基金
国家自然科学基金面上项目(81770246)
关键词
长链非编码核糖核酸H19
人主动脉平滑肌细胞
表型转化
动脉钙化
自噬
Log non-coding RNA H19
Human aortal vascular smooth muscle cells
Phenotype transformation
Atrial calcification
Autophagy
