生长相关蛋白-43及mRNA在大鼠损伤坐骨神经中的表达

目的探讨坐骨神经离断后,其远侧端组织中生长相关蛋白-43mRNA(GAP-43mRNA)及其蛋白表达的变化,为临床神经修复过程中更好地改善微环境提供实验依据。方法正常SD大鼠24只,行坐骨神经横断术,术后1,2,3,4周取坐骨神经远侧端组织,以WesternBlot和RT-PCR法检测GAP-43及其mRNA的表达。结果RT-PCR结果显示,正常大鼠坐骨神经组织中GAP-43mRNA表达量较低,损伤坐骨神经远侧端GAP-43mRNA的表达量明显增加,于损伤第2周时达高峰。采用抗GAP-43抗体,进行WesternBlot检测结果:正常坐骨神经43kDa处出现弱阳性反应的蛋白区带,损伤后坐骨神经远侧端43kDa处阳性反应条带着色明显加深,损伤后第3周时阳性反应着色最强。结论GAP-43及其mRNA在大鼠损伤坐骨神经远侧端组织中表达增强,其mRNA表达上调高峰在神经损伤后的第2周;而其蛋白合成增加最为明显的时间是在神经损伤后的第3周。

Expression of growth- associated protein(GAP- 43)and its mRNA in injured rat sciatic nerve

Aim To study the expression of growth associated protein (GAP 43) and its mRNA in distal stump of rat injured sciatic nerve and to provide experimental basis for improving microenviorment.Methods Western Blot and RT PCR were used to detect the different expression of GAP 43 and its mRNA in the distal stump of injured sciatic nerve.Results RT PCR result,the expression of GAP 43 mRNA was in low levels in the normal sciatic nerve and increased significantly after nerve injury,and reached climax at the second week after the sciatic nerve was injured.Western Blot results with anti GAP 43 antibody,the special positive band of 43 kDa protein was light in normal sciatic nerve and deep staining positive band of 43 kDa protein in the distal stump of injured sciatic nerve, and reached the peak at the third week.Conclusion The expression of GAP 43 and its mRNA are increased significantly in distal stump of injured sciatic nerve.The up regulation peak of GAP 43 mRNA is at the second week,the peak of the protein synthesis is at the third week, and its up regulation is impressed by nerve injury.