反义大鼠凝血酶受体基因的构建及其对主动脉血管平滑肌细胞增生的影响

凝血酶激活凝血酶受体引起的一系列细胞事件和内源性PDGF-A，bFGF的产生可能是血管病变发生发展过程中平滑肌细胞增生的重要因素。为进一步阐明血管损伤后平滑肌细胞增生的机理及探索有效治疗途径，针对凝血酶受体基因中一段包括转录起始点，翻译起始点，凝血酶结合和切割位点的序列作为靶序列，构建了凝血酶受体反义RNA重组表达载体pLXSN／ATR，并将其导入凝血酶刺激的动脉平滑肌细胞。结果表明重组基因转染能抑制凝血酶对大鼠平滑肌细胞DNA合成的刺激作用。提示序列特异的凝血酶受体反义重组基因的表达可以抑制凝血酶受体介导的血管平滑肌细胞增生。

Construction of antisense rat thrombin receptor gene and its effect on proliferation of rat aortic smooth muscle cells

Thrombin-induced receptor activation which leads to a number of intracellular signaling events as well asstimulation of endogenous PDGF-A and bFGF production may be important to the induction of proliferation of arterialsmooth muscle cells in the development of vascular disease. To obtain a better understanding of the mechanismsunderlying ASMC prolifeation after arterial injury and explore an effective therapy approach, a plasmid (pLXSN/ATR),which expresses antisense thrombin receptor (ATR) mRNA directed against the sequences of translation initiationregion, thrombin cleavage and binding sites of the thrombin receptor, was constructed and transfected into thecultured RASM. The results showed that 3H-TdR incorporation in the transfected RASM was inhibited with recombi-nant pLXSN/ATR. This indicated that specific inhibition of thrombin receptor-mediated ASMC proliferation waspossible using a receptor specific antisense DNA.