基于miR-27a介导的PCSK9/ABCA1途径探讨补肾降脂方抑制动脉粥样硬化的效应机制

Atherosclerosis is the common pathological basis for the occurrence of cardiac and cerebrovascular events. Reverse cholesterol transport plays an important role in the formation and development of AS. Recent studies have shown that miR-27a can be targeted to regulate the PCSK9/ABCA1 pathway that is closely related to reverse cholesterol transport. Our previous study confirmed the curative effect of Bushen jiangzhi Decoction on atherosclerosis, but its mechanism needs to be further studied. The project intends to adopt the whole animal experiment with in vitro cytology, using luciferase reporter gene analysis, gene silencing / expression, real-time fluorescence quantitative PCR, Western blot and other molecular biological methods, combined with pathological observation and bioinformatics technology, in order to clarify the scientific hypothesis of "antagonism of miR-27a and targeted regulation of the PCSK9/ABCA1 pathway, promoting reverse cholesterol transport may be an important mechanism of Bushen Jiangzhi Decoction inhibited AS. All that for providing the multi-target and deep-level experimental evidence for treatment of AS related diseases from the kidney.

动脉粥样硬化（Atherosclerosis, AS）是心、脑血管事件发生的共同病理基础，胆固醇逆向转运在AS形成、发展过程中发挥关键作用。新近有研究表明，miR-27a可靶向调控与胆固醇逆向转运密切相关的PCSK9/ABCA1途径。我们前期研究证实补肾降脂方抗AS疗效肯定，但其作用机制有待深入研究。本项目拟采用整体动物实验结合体外细胞学研究，应用荧光素酶报告基因分析、基因沉默/过表达、实时荧光定量PCR、Western blot等分子生物学方法，结合组织病理检测、生物信息学等技术，以期阐明“拮抗miR-27a、靶向调控PCSK9/ABCA1途径，促进胆固醇逆向转运可能是补肾降脂方抑制AS的重要作用机制”的科学假说，为从补肾论治AS相关疾病提供多靶点、深层次的实验依据。

动脉粥样硬化（Atherosclerosis, AS）多发于中老年人，系心脑血管事件之病理基础，其本虚标实的病机特点为业界所公认，立足以肾为本、从肾论治AS之补肾降脂方已证实其有效性。基于胆固醇逆向转运及其自噬在AS形成、发展中发挥重要作用，miR-27a调控与胆固醇逆向转运密切相关的PCSK9/ABCA1途径，本项目以整体动物实验结合体外细胞学研究（高脂饮食喂饲apoE-/-小鼠复制AS模型/ox-LDL诱导RAW264.7小鼠巨噬细胞源性泡沫细胞），初步阐明了补肾降脂方通过PCSK9调控CD36、PPARγ-LXRα-ABCA1通路以及mTOR通路调控自噬干预AS的效应机制；探究了基于自噬的Quercetin调控MST1介导的RAW264.7小鼠巨噬细胞源性泡沫细胞以及apoE-/-小鼠AS的相关机制；完成Fisetin通过CKIP-1/REGγ途径保护ox-LDL诱导的RAW264.7小鼠巨噬细胞源性泡沫细胞脂质蓄积和衰老的机制研究以及Fisetin调控PCSK9、LOX-1干预apoE-/-小鼠AS的机制研究。以上较为深入地解析了补肾降脂方及其中药单体Quercetin、Fisetin通过调控胆固醇逆向转运、自噬及其相关信号通路进而有效干预AS的相关机制，可为中医药防治AS相关疾病提供多靶点、深层次的科学依据，彰显中医药防抗AS丰富的科学内涵。