长链非编码RNA-p21诱导细胞凋亡在动脉粥样硬化中的作用-东南大学学报医学版

长链非编码RNA-p21诱导细胞凋亡在动脉粥样硬化中的作用

单位：1. 石河子大学医学院第一附属医院心胸外一科, 新疆石河子 832008;
2. 山东省潍坊市市直机关医院超声科, 山东潍坊 261061

分类号：R541.4

出版年·卷·期（页码）：2017·36·第五期（816-821）

摘要：

目的：探讨lincRNA-p21通过调控细胞凋亡参与冠心病的发生发展过程，及其对动脉粥样硬化的影响。方法：选取我院50例冠心病患者，用ElISA方法检测患者血清中lincRNA-p21含量和凋亡蛋白Bcl-2含量。提取大鼠心肌内皮细胞制备大鼠动脉硬化模型，过表达或抑制lincRNA-p21后，CCK-8法检测细胞存活率；构建lincRNA-p21质粒注射进入小鼠颈动脉损伤模型，HE染色观察小鼠颈动脉内膜新生。Caspase-3活性检测试剂盒检测内皮细胞凋亡蛋白Caspase-3活性；Western blot法检测凋亡蛋白Bcl-2和BAX蛋白表达水平。结果：冠心病患者血清内lincRNA-p21表达显著下降，差异有统计学意义（P<0.05）。大鼠原代内皮细胞中过表达lincRNA-p21显著降低细胞存活率，显著上升Caspase-3活性，显著增加BAX表达，显著下降BCL-2表达；lincRNA-p21抑制组细胞存活率明显增高，Caspase 3活性显著下降，促凋亡蛋白BAX水平显著降低，抗凋亡蛋白BCL-2水平显著增加，差异有统计学意义（P<0.05）。小鼠体内局部沉默lincRNA-p21后，血管内/中膜厚度比明显增加，Caspase-3活性明显降低，Bcl-2蛋白表达增加且BAX蛋白表达降低，差异有统计学意义（P<0.05）。结论：lincRNA-p21可通过调控细胞凋亡参与冠心病的发生发展过程。

Objective:To investigate the effect of lincRNA-p21 on the development of coronary heart disease (CHD) via regulating apoptosis. Methods:Fifty patients with coronary heart disease were randomly divided into two groups. The content of lincRNA-p21 and the content of apoptotic protein BCL-2 were detected through ELISA method. Rat cardiac endothelial cells were extracted to prepare for rat arteriosclerosis model with overexpression or inhibition of lincRNA-p21, followed by the detection of cell viability using CCK-8 method. LincRNA-p21 plasmid was constructed and implanted into the mice carotid artery injury model, Carotid artery neointima in mice was detected by HE staining. Caspase 3 activity assay kit was used to detect the expression of Caspase 3 in endothelial cells. The expression of BCL-2 and BAX protein was detected by Westernblot. Result:The expression of lincRNA-p21 in serum of patients with coronary heart disease was significantly decreased(P<0.05). The cell viability of lincRNA-p21 overexpression in rat primary endothelial cells was significantly reduced, Caspase 3 activity was significantly increased,BAX protein expression was significantly increased,and BCL-2 protein expression was significantly decreased. LincRNA-p21 inhibation was significantly increased the cell survival, Caspase 3 activity was significantly decreased, the level of pro-apoptotic protein BAX was significantly decreased, anti-apoptotic protein BCL-2 levels increased significantly, the difference was statistically significant(P<0.05). Intravascular/medial thickness ratio was increased significantly after mice localized with lincRNA-p21 inhibition, Caspase 3 activity was significantly decreased, Bcl-2 protein expression was increased and BAX protein expression was decreased, the difference was statistically significant (P<0.05). Conclusion:lincRNA-p21 could be involved in the development of coronary heart disease by regulating apoptosis.

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