循环外泌体中microRNA30d上调mGluR5促冠状动脉内皮细胞凋亡的机制研究

The pathogenesis of coronary heart disease (CHD) remains unclear。 The prevailing statement is the theory of endothelial injury response. Microarray analysis showed that MicroRNA30d was highly expressed in CHD via microarray. And our previous results of experiments showed that the expression of circulating exosomes- derived microRNA 30d was increased in CHD, positively correlated with the expression of endothelin and sVCAM-1. Bioinformatic analysis found that the target gene of microRNA30d was mGluR5, possibly regulated by PKC pathway. Pre-experiment confirmed that microRNA30d could up-regulate mGluR5, promoting HCACE apoptosis, but the mechanism has not been clear. Scientific hypothesis: Overexpressed circulating exosomes- derived microRNA 30d upregulated mGluR5 in CHD, via the PKC pathway leading to HCACE apoptosis. The aim of this study was to investigate the effect of microRNA30d, microRNA30d-mGluR5 and PKC pathway on HCACE apoptosis, explore the mechanism of classical theory of endothelial cell injury, and provide a new idea for targeted therapy towards CHD.

冠心病的发病机制尚不清楚，目前较公认的是“内皮损伤反应学说”。microRNA30d是我们采用芯片筛选到，高表达于冠心病、功能未知的microRNA。前期研究发现：冠心病循环外泌体microRNA30d增高，与内皮素和sVCAM-1正相关；生物信息学提示microRNA30d的靶基因为mGluR5，并可通过PKC通路发生作用；预实验证实microRNA30d可上调mGluR5，促HCACE凋亡，但机制不清楚。故提出科学假说：冠心病循环外泌体microRNA30d高表达上调mGluR5，通过PKC通路导致HCACE凋亡。本研究拟从临床、动物和细胞三个层面，用过表达与抑制microRNA30d，以microRNA30d- mGluR5、PKC通路为机制主线，论证循环外泌体microRNA30d在HCACE中的调控作用，对经典内皮细胞损伤机制予以补充，并为冠心病靶向治疗提供新的思路。

冠心病的发病机制尚不清楚，目前较公认的是“内皮损伤反应学说”。microRNA30d是我们采用芯片筛选到，高表达于冠心病、功能未知的microRNA。通过对288例临床数据证实，冠心病循环外泌体microRNA30d增高，与内皮素及sVCAM-1正相关。生物信息学预测提示microRNA30d的靶基因为mGluR5，并可能通过PKC通路发生作用。通过双荧光素酶检测证实microRNA30d的靶基因确实为mGluR5。细胞实验证实microRNA30d可通过mGluR5-PLC-PKC信号通路，促HCACE凋亡；并进一步证实microRNA30d作用与ox-LDL相似，且与ox-LDL对内皮细胞有协同作用。通过动物实验证实，高脂动物组microRNA30d也增高，且通过mGluR5-PLC-PKC信号通路，促进内皮细胞凋亡。证实冠心病中microRNA30d增高与内皮细胞损伤确实有关联，为冠心病靶向治疗提供新的思路。