Kindlin-2 RNA干扰减轻大鼠颈动脉内膜增生

Kindlin-2RNA interference alleviates intimal hyperplasia of rat carotid arteries

目的观察Kindlin-2 RNA干扰对球囊损伤的大鼠颈动脉内膜增生的影响。方法构建并制备Kindlin-2 siRNA和阴性对照慢病毒载体并感染球囊损伤的大鼠颈动脉,1周和4周后取损伤的颈动脉,实时定量PCR检测Kindlin-2 mRNA的表达,HE染色评估术后4周颈动脉内膜增生情况,并行免疫荧光或免疫组化检测颈动脉中Kindlin-2、α-actin、PCNA和β1整合素的表达,Masson染色了解颈动脉中胶原纤维合成情况。结果 Kindlin-2RNA干扰能明显降低1周时颈动脉中Kindlin-2 mRNA的表达水平(P<0.05),并显著抑制4周后颈动脉的内膜增生(P<0.05)。与阴性对照组相比,Kindlin-2 siRNA组术后4周颈动脉中Kindlin-2、α-actin、PCNA和β1整合素的表达水平均明显降低,颈动脉内膜和中膜胶原纤维的合成也减少。结论 Kindlin-2 RNA干扰可能通过抑制血管平滑肌细胞增殖、迁移和细胞外基质合成减轻血管内膜增生。

Aim To observe the effects of Kindlin-2 RNA interference on intimal hyperplasia in the balloon injured rat carotid arteries. Methods Kindlin-2 siRNA and negative control lentiviral vectors were constructed and produced,then balloon injured rat carotid arteries were infected with lentivirus. Carotid arteries were collected at 1 and 4 weeks after operation. The expression level of Kindlin-2 mRNA in the rat carotid arteries was detected by real-time quantitative PCR technique. The degree of intimal hyperplasia 4 weeks after balloon injury was assessed by HE staining. The immunofluorescence and histochemistry were used to detect the protein expression of Kindlin-2,α-actin,PCNA and β1-integrin in the carotid arteries. The synthesis of collagen fibers in the carotid arteries was investigated by Masson staining.Results Kindlin-2 RNA interference can significantly reduce the Kindlin-2 mRNA expression level in the rat carotid arteries at 1 week( P<0.05),and significantly inhibited intimal hyperplasia of carotid arteries 4 weeks after balloon injury( P<0.05). Compared with the negative control group,the protein expression levels of Kindlin-2,α-actin,PCNA and β1-integrin were significantly decreased in the carotid arteries of the Kindlin-2 group 4 weeks after operation. The synthesis of collagen fibers in the carotid arteries intima and media was also reduced. Conclusion Kindlin-2 RNA interference may inhibit vascular smooth muscle cell proliferation,migration and extracellular matrix synthesis to alleviate intimal hyperplasia.