反义凝血酶受体基因序列对大鼠血管平滑肌细胞增殖的影响  

Antisense Sequences of Thrombin Receptor Arrest of rat Vascular Smooth Muscle Cell Proliferation

作　　者：王启贤[1] 肖丽梅[1] 李继梅[2] 郝萍[2] 杨源[1] 黄韬[3] 肖践明[1] 杨西云[1] 山路[1] 唐睿珠[2] 

机构地区：[1]昆明医学院附属第一医院心内科,云南省昆明市650032 [2]昆明医学院附属第一医院临床实验中心,云南省昆明市650032 [3]昆明医学院附属第一医院胸外科,云南省昆明市650032

出　　处：《中国动脉硬化杂志》2004年第5期519-523,共5页Chinese Journal of Arteriosclerosis

基　　金：云南省应用基础研究基金 ( 1999C0 0 67M)资助

摘　　要：为了研究凝血酶受体在血管平滑肌细胞增埴中的作用 ,探讨反义凝血酶受体基因序列对大鼠血管平滑肌细胞增殖的影响 ,通过培养SD大鼠胸主动脉血管平滑肌细胞 ,以3 H TdR掺入率作为评价反义凝血酶受体基因序列抑制血管平滑肌细胞增殖的指标 ;用反转录聚合酶链反应检测反义凝血酶受体基因序列抑制血管平滑肌细胞凝血酶受体mRNA的表达 ;用Western blot检测反义凝血酶受体基因序列抑制血管平滑肌细胞凝血酶受体蛋白质的表达 ;用3 H 肌醇掺入率检测反义凝血酶受体序列抑制血管平滑肌细胞磷酸肌醇代谢的影响。结果发现 ,反义凝血酶受体基因序列明显抑制血管平滑肌细胞的增殖 (与对照组相比 ,P <0 .0 5 ) ;明显抑制血管平滑肌细胞的凝血酶受体mRNA和蛋白的表达 ;明显抑制了血管平滑肌细胞磷酸肌醇代谢。提示反义凝血酶受体基因序列明显抑制血管平滑肌细胞的增殖 ;凝血酶受体反义序列明显抑制血管平滑肌细胞的增殖是通过抑制凝血酶受体基因的表达 (特别是通过抑制DNA、mRNA和蛋白的表达 ) 。Aim To study the influence of thrombin receptor on the proliferation of rat vascular smooth muscle cells,probe into mechanism of antisense sequence of thrombin receptor restrained the proliferation of rat vascular smooth cells. Methods Take the incoporation of 3H-thymidine as the target to evaluate vascular smooth muscle cells proliferation; Antisense sequence of thrombin receptor restrained the expression of mRNA of thrombin receptorin vascular smooth cells was detected by RT-PCR; Antisense sequence of thrombin receptor restrained the expression of protein of thrombin receptor was detected by Western-blot; The influence of antisense sequence of thrombin receptor on phosphate inositol of vascular smooth cells was detected by incoporation of 3H-inositol. Results Antisense sequence of thrombin receptor restrained vascular smooth muscle cells proliferation marketedly (compared with control P<0.05),restrained expression of mRNA and protein of thrombin receptor marketedly;Also restrained metabolism of phosphate inositol in vascular smooth cells. Conclusion Antisense sequence of thrombin receptor restrained SD rat vascular smooth muscle cells proliferation marketedly; Antisense sequence of thrombin receptor restrained vascular smooth muscle cells proliferation which could arrested gene expression of thrombin receptor,restrained signal trasduction of vascular smooth muscle cells.

关 键 词：病理学与病理生理学 血管平滑肌细胞增殖 凝血酶 受体 凝血酶 基因序列反义技术 动脉粥样硬化 

分 类 号：R363[医药卫生—病理学]