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.