p27kip1和Skp2在损伤后大鼠坐骨神经中的表达变化

为了探讨损伤后周围神经p27kip1和S期激酶相关蛋白2(Skp2)的定位表达和变化,本实验将成年SD大鼠随机分为正常对照组、夹伤组和切断组,运用Western blot结合免疫组织化学及免疫荧光双标,在大鼠坐骨神经损伤时,对p27kip1和Skp2表达的影响进行了研究。结果表明:(1)坐骨神经夹伤后,p27kip1蛋白表达先逐渐下降,后又逐渐上升;坐骨神经切断后,远侧段p27kip1蛋白表达持续下降,而近侧段p27kip1蛋白表达在切断后6h明显下降,后又逐渐升高至正常水平,而Skp2表达变化与之相反;(2)免疫组织化学染色结果显示,坐骨神经切断后1w,远侧段从断端到末端,p27kip1阳性信号逐渐增加,而Skp2阳性信号逐渐减弱;(3)免疫荧光双标显示,正常和损伤坐骨神经的雪旺氏细胞中都有p27kip1和Skp2表达。以上结果提示:周围神经损伤后影响雪旺氏细胞中p27kip1和Skp2的表达变化,为进一步研究它们在周围神经损伤和修复中的作用机制奠定基础。

EXPRESSION OF p27kip1 AND Skp2 IN INJURED SCIATIC NERVE OF ADULT RATS

To investigate the expression of p27kip1 and S-phase kinase associated protein 2 (Skp2) in injured sciatic nerve, the adult rats were randomly divided into control group,sciatic nerve crush group and sciatic nerve transection group. The effect of sciatic nerve injury on the expression of p27kip1 and Skp2 was studied by Western blot, immunohistochemistry and double immunofluscent labelling. The results showed that: (1) the expression of p27kip1 and Skp2 were detected in sciatic nerve, the level of p27kip1 was declined after sciatic nerve crush, then it was rescued. While it was decreasd continually in the distal stump following sciatic nerve transection. But in the proximal stump, the expression of p27kip1 was declined as early as 6 h after sciatic nerve transection, then it was increased gradually to normal level. And the changes of p27kip1 and Skp2 in rat sciatic nerve were inversely correlated; (2) by immunohistochemistry, it was found that at 1 week post transection ,from the injured site to the end of the distal stump, the positive signal of p27kip1 was regularly increased. Inversely, the positive signal of Skp2 was regularly decreased;(3) double immunolabelling showed that both p27kip1 and Skp2 were expressed in Schwann cells in normal and injured sciatic nerve. These results suggested that sciatic nerve lesion could affect the expression level of p27kip1 and Skp2 in Schwann cells,which is important for the further research on the effect and mechanism of them on peripheral nerve injury.