RNA干扰基因沉默单核细胞趋化蛋白-1干预动脉粥样硬化早期形成的研究

Experimental studies on anti-atherosclerosis by RNA interference mediated silencing of monocyte chemotactic protein-1 expression

目的观察单核细胞趋化蛋白-1（MCP-1）、小干扰RNA（siRNA）、转染兔颈动脉粥样硬化（AS）模型对局部MCP-1表达及AS早期形成的影响。方法建立28只雄性兔颈动脉AS模型，将MCP-1siRNA真核表达质粒通过局部定位转染的方法至病变血管，病理形态学观察血管壁中泡沫细胞浸润及内膜增厚，免疫组织化学、逆转录-聚合酶链反应（RT—PCR）、Westernblot检测局部MCP-1表达。结果模型对照组、空质粒组、RNA干扰（RNAi）组镜下内膜／中膜厚度（I：M）分别为5．55、5．27、1．46。免疫组织化学染色显示RNAi组着色较浅。RT—PCR及Westernblot结果显示：RNAi组与模型对照组、空质粒组比较，局部MCP-1表达降低。结论MCP-1siRNA转染抑制了局部MCP-1表达，延缓了AS的发展，减轻了病变程度。

Objective To identify the effects of in vivo transfeetion of monoeyte chemotactic protein-1 （ MCP-1 ） small interference RNA （siRNA） to atheroselerosis model on local expression of MCP-1 and the development of atheroselerosis. Methods Carotid atherosclerosis model was created on 28 New Zealand white rabbits, siRNA expressing vector was transfected to the blood vessels. The carotid artery was processed for morphologie evaluation. The local expression of MCP-1 was detected by immunohistoehemistry, RT-PCR and Western blot. Results The intima：media in thickness in RNA interference, control and blank plasmid groups was 5.55,5.27 ,and 1.46 respectively. Immunohistoehemistry showed,as compared with control and blank plasmid groups, the positie staining of MCP-1 was decreased in RNA interference group. RT-PCT and Western blot revealed that MCP-1 expression was reduced in RNA interference group as compared with model and blank plasmid groups. Conclusion The expression of MCP-1 was successfully inhibited by transfecting MCP-1 siRNA expression plasmid to the carotid artery, and the progression of atherosclerosis was limited by RNA interference-mediated silencing of MCP-1 expression.