MicroRNA-140-3p下调促进自噬调控下肢动脉硬化闭塞平滑肌细胞功能研究

Misregulation of MicroRNAs may cause proliferation and migration of artery smooth muscle cell (ASMCs) and play pivotal roles in the genesis and process of arteriosclerosis obliterans (ASO), but the funtion of changed microRNA remains unclear. We found that miR-140-3p was significantly downregulated in ASO media, and upregulation of miR-140-3p markedly attenuated ASMCs proliferation. However, the exact role and underlying mechanisms of how miR-140-3p regulates ASMCs behavior in ASO remains unknown. Further study showed that it likely associated with Lysosome-associated membrane protein (LAMP)-2, an important protein in autophagic flux. In the present study, we intend to use methods such as miR-140-3p mimic and inhibitor transfection, dual-luciferase assay, lentivirus mediated overexpression, and siRNA to prove that miR-140-3p regulates ASMCs behavior via LAMP-2, and to explore the role of miR-140-3p in ASO. The present study would shed light on the possible role and mechanism of miR-140-3p/LAMP-2 in modulating ASMCs behavior in ASO, and provide solid evidence for proving miR-140-3p/LAMP-2 as a new potential therapeutic approach for preventing and treating ASO.

我们发现microRNA表达失调引起动脉平滑肌细胞(ASMCs)增殖迁移失控是导致下肢动脉硬化闭塞(ASO)的重要原因，但ASO中表达失调microRNA的功能仍不清楚。我们发现ASO的ASMCs中miR-140-3p表达下调，上调则明显抑制ASMCs增殖，然而其对ASMCs迁移、凋亡等的影响以及内在调控机制仍然未知，深入分析发现可能与自噬相关蛋白溶酶体关联膜蛋白(Lysosome-associated membrane protein, LAMP)-2有关。本研究拟采用microRNA转染、双荧光素酶报告基因、慢病毒表达、siRNA等方法，旨在获得miR-140-3p通过LAMP-2调控ASMCs功能的证据，探讨其在ASO发病中的作用。本项目将揭示ASO发病中miR-140-3p/ LAMP-2调控ASMCs生物学行为变化的新机制，为确立其作为干预ASO的新途径提供依据。

我们发现microRNA表达失调引起动脉平滑肌细胞(ASMCs)增殖迁移失控是导致下肢动脉硬化闭塞(ASO)的重要原因，但ASO中表达失调microRNA的功能仍不清楚。我们发现ASO的ASMCs中miR-140-3p表达下调，上调则明显抑制ASMCs增殖，然而其对ASMCs迁移、凋亡等的影响以及内在调控机制仍然未知，深入分析发现可能与自噬相关蛋白溶酶体关联膜蛋白(Lysosome-associated membrane protein, LAMP)-2有关。在实验进展过程中，我们未发现miR-140-3p与LAMP-2的相关性，但是发现了miR-130a更具有实验意义，我们继而开始深入研究miR-130a的相关机制。本研究拟采用microRNA转染、双荧光素酶报告基因、慢病毒表达、siRNA等方法，旨在获得miR-130a通过凋亡调控ASMCs功能的证据，探讨其在ASO发病中的作用。本项目将揭示ASO发病中miR-130a/凋亡调控ASMCs生物学行为变化的新机制，为确立其作为干预ASO的新途径提供依据。