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目的 :将构建的AT2 RcDNA重组质粒转染到培养的大鼠血管平滑肌细胞 ,为研究AT2 R对血管平滑肌细胞生长的影响提供实验基础。方法 :扩增、纯化重组质粒后 ,用脂质体介导法将重组质粒转染入培养的大鼠血管平滑肌细胞 ,分别用AT2 RmRNA原位杂交、AT2 R抗体免疫组化的方法从RNA和蛋白水平检测转染效果。结果 :原位杂交显示部分细胞浆棕黄色阳性反应 ,显示转染率为 9% ,免疫组化结果显示部分血管平滑肌细胞膜上有AT2 R蛋白存在。结论 :成功将AT2 RcDNA重组质粒转染到培养的大鼠血管平滑肌细胞中。转染后AT2 R能在大鼠主动脉平滑肌细胞表达的现象为进一步研究AT2 R对成年大鼠血管平滑肌细胞生长、凋亡的影响提供了实验基础
OBJECTIVE: Transfection of recombinant AT2 RcDNA into cultured rat vascular smooth muscle cells provides the experimental basis for studying the effect of AT2 R on the growth of vascular smooth muscle cells. Methods: The recombinant plasmids were amplified and purified. The recombinant plasmids were transfected into cultured rat vascular smooth muscle cells by liposome-mediated method. The antisense oligonucleotides of AT2 RmRNA and AT2 R antibody Protein levels were tested for transfection efficiency. Results: In situ hybridization showed some cytoplasm brown positive reaction, showing that the transfection rate was 9%. Immunohistochemistry showed that some of the vascular smooth muscle cells had AT2 R protein present. Conclusion: AT2 RcDNA recombinant plasmid was successfully transfected into cultured rat vascular smooth muscle cells. The expression of AT2 R in rat aortic smooth muscle cells after transfection provides the experimental basis for further study on the effect of AT2 R on the growth and apoptosis of rat vascular smooth muscle cells