Investigation of highly expressed PCSK9 in atherosclerotic plaques and ox-LDL-induced endothelial cell apoptosis.
Investigation of highly expressed PCSK9 in atherosclerotic plaques and ox-LDL-induced endothelial cell apoptosis.
复制标题
DOI：
10.3892/mmr.2017.6803
复制
发表时间：
2017-08
影响因子：
3.4
通讯作者：
Li G
中科院分区：
文献类型：
--
作者：
Li J;Liang X;Wang Y;Xu Z;Li G
本研究旨在探讨枯草素转化酶（protein converting ase subtilisin/ keexin type 9, PCSK9）对动脉粥样硬化（atherosclerosis， AS）的直接毒性及其与内皮细胞凋亡的关系。载脂蛋白E−/−小鼠随机分为对照组和实验组。对照组饲喂正常饮食，试验组饲喂高脂饮食。20周后，分离主动脉并剥离。苏木精和伊红染色，免疫组织化学分析。将人脐静脉内皮细胞与不同浓度的氧化低密度脂蛋白（ox-LDL）孵育不同时间。流式细胞术检测细胞凋亡率。采用Western blotting和逆转录-定量聚合酶链反应检测PCSK9、b细胞淋巴瘤2 （Bcl-2）、Bcl-2样蛋白4 （Bax）和caspase-3的表达。用慢病毒转染法将短发夹RNA-PCSK9转染内皮细胞。检测PCSK9、Bax、Bcl-2、caspase-3和丝裂原活化蛋白激酶（MAPK）通路蛋白的表达水平。高脂组成功建立as模型，PCSK9在as斑块中高表达。ox-LDL诱导细胞凋亡，PCSK9 mRNA和蛋白水平升高。shRNA-PCSK9下调PCSK9 mRNA和蛋白水平。缺乏PCSK9可显著抑制促凋亡蛋白的表达，促进抗凋亡蛋白的表达。此外，shRNA-PCSK9还改变了p38和c-Jun n端激酶的磷酸化。shrna靶向PCSK9可能通过MAPK信号抑制AS内皮细胞凋亡，为了解AS的机制和治疗提供了新的方向。
The present study aimed to explore the direct toxicity of proprotein convertase subtilisin/kexin type 9 (PCSK9) to atherosclerosis (AS) and its association with apoptotic endothelial cells. Apolipoprotein E−/− mice were randomly divided into two groups, control and experimental. The control group was administered a normal diet and the experimental group was administered a high-fat diet. After 20 weeks, the aorta was isolated and dissected. Hematoxylin and eosin staining, and immunohistochemical analysis were performed. Human umbilical vein endothelial cells were incubated with varied concentrations of oxidized low-density lipoprotein (ox-LDL) for different times. The apoptotic rate was detected by flow cytometry. Western blotting and reverse transcription-quantitative polymerase chain reaction analysis were conducted to detect the expression of PCSK9, B-cell lymphoma 2 (Bcl-2), bcl-2-like protein 4 (Bax) and caspase-3. Short hairpin (sh) RNA-PCSK9 was transfected into endothelial cells using lentiviral transfection. The expression levels of PCSK9, Bax, Bcl-2, caspase-3 and the mitogen-activated protein kinase (MAPK) pathway proteins were detected. The high-fat group was successfully established as an AS model and PCSK9 was highly expressed in the AS plaque. Treatment with ox-LDL induced apoptosis and increased mRNA and protein levels of PCSK9. PCSK9 mRNA and proteins levels were downregulated by shRNA-PCSK9. The deficiency of PCSK9 markedly inhibited the expression of pro-apoptotic proteins and promoted anti-apoptotic proteins. In addition, phosphorylation of p38 and c-Jun N-terminal kinases was altered by shRNA-PCSK9. Targeting of PCSK9 by shRNA-PCSK9 may repress endothelial cell apoptosis through MAPK signaling in AS, providing a novel direction for understanding the mechanism and treatment of AS.
登录
查看更多内容
影响因子：
11.4
作者：
Elbashir, SM;Martinez, J;Tuschl, T
通讯作者：
Tuschl, T
DOI：
10.2147/dddt.s36984
发表时间：
2013
期刊：
Drug design, development and therapy
影响因子：
--
作者：
Poirier S;Mayer G
通讯作者：
Mayer G
影响因子：
37.8
作者：
Kwon, Gina P.;Schroeder, Jamie L.;Balaban, Robert S.
通讯作者：
Balaban, Robert S.
影响因子：
7.7
作者：
Constantinides, Alexander;Kappelle, Paul J. W. H.;Dullaart, Robin P. F.
通讯作者：
Dullaart, Robin P. F.
影响因子：
158.5
作者：
Roth, Eli M.;McKenney, James M.;Stein, Evan A.
通讯作者：
Stein, Evan A.