Moderate Autophagy Inhibits Vascular Smooth Muscle Cell Senescence to Stabilize Progressed Atherosclerotic Plaque via the mTORC1/ULK1/ATG13 Signal Pathway.
Moderate Autophagy Inhibits Vascular Smooth Muscle Cell Senescence to Stabilize Progressed Atherosclerotic Plaque via the mTORC1/ULK1/ATG13 Signal Pathway.
复制标题
中度自噬通过 mTORC1/ULK1/ATG13 信号通路抑制血管平滑肌细胞衰老以稳定进展的动脉粥样硬化斑块
DOI：
10.1155/2017/3018190
复制
发表时间：
2017
影响因子：
--
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Luo Z;Xu W;Ma S;Qiao H;Gao L;Zhang R;Yang B;Qiu Y;Chen J;Zhang M;Tao B;Cao F;Wang Y
为了研究雷帕霉素诱导的自噬对动脉粥样硬化斑块形成的影响，我们建立了高脂高胆固醇饮食（HFD）诱导ApoE−/−小鼠动脉粥样硬化模型，持续16周。雷帕霉素（Rapamycin）和3-甲基腺苷（MA）分别作为自噬诱导剂和抑制剂。采用HE和油红O染色检测主动脉斑块区。分别采用免疫组织化学染色法观察斑块的含量。与对照组和3-MA组相比，雷帕霉素可以抑制动脉粥样硬化的进展。雷帕霉素能相对增加胶原含量和a-SMA分布，减少坏死核心面积。然后用MOVAS和ox-LDL培养72 h，体外诱导平滑肌源性泡沫细胞模型。雷帕霉素与3-MA分别联合培养。采用流式细胞术和SA-β-Gal染色法检测VSMCs的存活和衰老情况。Western blot分析蛋白表达水平。我们发现，与对照组和3-MA组相比，雷帕霉素可以促进ox- ldl诱导的VSMCs自噬存活，减轻细胞衰老。Western blot分析显示，雷帕霉素可上调ULK1、ATG13蛋白表达，下调mTORC1和p53蛋白表达。
In order to investigate the effects of autophagy induced by rapamycin in the development of atherosclerosis plaque we established murine atherosclerosis model which was induced in ApoE−/− mice by high fat and cholesterol diet (HFD) for 16 weeks. Rapamycin and 3-Methyladenine (MA) were used as autophagy inducer and inhibitor respectively. The plaque areas in aortic artery were detected with HE and Oil Red O staining. Immunohistochemical staining were applied to investigate content of plaque respectively. In contrast to control and 3-MA groups, rapamycin could inhibit atherosclerosis progression. Rapamycin was able to increase collagen content and a-SMA distribution relatively, as well as decrease necrotic core area. Then we used MOVAS and culture with ox-LDL for 72 h to induce smooth muscle-derived foam cell model in vitro. Rapamycin and 3-MA were cultured together respectively. Flow cytometry assay and SA-β-Gal staining experiments were performed to detect survival and senescence of VSMCs. Western blot analysis were utilized to analyze the levels of protein expression. We found that rapamycin could promote ox-LDL-induced VSMCs autophagy survival and alleviate cellular senescence, in comparison to control and 3-MA groups. Western blot analysis showed that rapamycin could upregulate ULK1, ATG13 and downregulate mTORC1 and p53 protein expression.