反义Smad3阻断TGF-β_1信号转导对血管平滑肌细胞增殖能力的影响

Effects of signal transduction interruption of transforming growth factor-β_1 by anti-Smad3 on proliferation of vascular smooth muscle cells

目的观察反义Smad3腺病毒载体转染增生型血管平滑肌细胞(VSMC)后对其增殖能力的影响。方法构建大鼠颈总动脉急性球囊损伤模型。取损伤处血管组织,组织块贴壁法培养增生型VSMC,反义Smad3腺病毒载体转染(实验组)。以空载病毒作为阴性对照组,以无病毒载体作为空白对照组。抽提转染后细胞总RNA,RT-PCR检测转染后各组Smad3 mRNA表达;MTT比色法检测转染后细胞增殖能力的变化。结果与阴性对照组和空白对照组比较,转染后实验组Smad3 mRNA表达明显减少;转染后第2、3、5天,实验组细胞增殖能力明显降低(P<0.05),转染后第7天各组细胞增殖能力差异无统计学意义(P>0.05)。结论反义Smad3腺病毒载体转染对增生型VSMC具有增殖抑制作用,为通过基因修饰阻断TGF-β1信号转导从而预防增生性血管病变奠定了基础。

Objective To observe the effects of anti-Smad3 adenovirus vector transfection on proliferation of proliferative vascular smooth muscle cells （ VSMCs）. Methods Rat models of acute balloon injury of common carotid artery were established. The injured vascular tissues were obtained, proliferative VSMCs were cultivated by tissue explants adherent method, and anti-Smad3 adenovirus vector transfection was performed （experiment group）. Virus without anti-Smad was served as negative control, and that without adenovirus vector was served as blank control. Total RNA of transfected cells was extracted, and the expression of Smad3 mRNA after transfection in each group was detected by RT-PCR. The changes in cell proliferation after transfection was determined by MTT. Results The expression of Smad3 mRNA in experiment group was significantly lower than that in negative control group and blank control group. Cell proliferation of experiment group was significantly decreased on day 2, 3 and 5 after transfection （P 〈 0.05）, while there was no significant difference in cell proliferation among each group on day 7 after transfection （P 〉 0. 05）. Conclusion Anti-Smad3 adenovirus vector transfection may inhibit the proliferation of proliferative VSMCs, which lays a foundation for the signal transduction interruption of TGF-β1 by gene modification in the prevention of proliferative vascular diseases.