核糖核酸干涉血小板源性生长因子表达抑制视网膜色素上皮细胞增生的实验研究  

The experimental study of RNA interference in inhibition of platelet derived growth factor expression and retinal pigment epithelial cells proliferation

作　　者：孟祥俊[1] 周明[1] 刘豪杰[1] 胥利平[1] 

机构地区：[1]大连大学附属中山医院眼科,辽宁116001

出　　处：《中华眼外伤职业眼病杂志》2015年第9期657-661,共5页Chinese Journal of Ocular Trauma and Occupational Eye Disease

基　　金：辽宁省自然科学基金（20102007）

摘　　要：目的探讨核糖核酸（RNA）在基因水平上干涉血小板源性生长因子（PDGF）的转录与表达，从而抑制视网膜色素上皮细胞（RPE）增生的机制，以期对外伤性PVR的防治提供帮助。方法针对PDGF基因的小干扰核糖核酸（siRNA）序列设计引物，体外转录、转染细胞，寻找具有高效干涉作用的siRNA序列，利用构建具有体内转录siRNA功能的慢病毒载体转染培养的RPE细胞，采用生长曲线及噻唑蓝比色法（MTT）检测PDGF siRNA-慢病毒抑制RPE细胞增生的作用，并用流式细胞仪检测细胞周期的变化。结果获得具有高效干涉PDGF表达的siRNA序列，构建出转录该siRNA序列的慢病毒载体，且转染RPE细胞后可抑制RPE细胞增生。结论利用RNA的干涉作用，在基因水平上抑制PDGF的转录与表达，从而抑制RPE细胞的增生。Objective To investigate the effect of RNA interference in transcription and expression of platelet derived growth factor （PDGF） and retinal pigment epithelial ceils （RPE） proliferation. Methods i According to the PDGF siRNA sequence the primers was designed and siRNA was obtained in vitro by transcription and then transfected cells. The siRNA sequences with high interference effect were looked for. The lentiviral vector with siRNA expression function in vivo was constructed and transfected into cultured RPE cells. The inhibition effect of PDGF siRNA on the proliferation of RPE cells was measured by the growth curve and MTT and the changes of cell cycles were decected by flow cytometry. Results SiRNA sequences with high interference in inhibition of the expression of PDGF were obtained, and the recombinant lentiviral vetor of siRNA sequence was constructed. SiRNA-cells which were transfected into RPE cells inhibited the proliferation of RPE cells. Conclusion Using the interference effect of RNA, the transcription and expression of PDGF was inhibited at the gene level, and the proliferation of RPE cells was inhibited.

关 键 词：视网膜色素上皮 血小板源性生长因子 增生性玻璃体视网膜病变 外伤性 核糖核酸干涉 

分 类 号：R774.1[医药卫生—眼科]