慢病毒介导的红色荧光蛋白基因在大鼠坐骨神经细胞的表达

Lentiviral vector-mediated transfection of RFP Gene into rat sciatic nerve cells

目的 研究慢病毒载体介导的红色荧光蛋白（RFP）基因在坐骨神经雪旺细胞体内转染情况.方法 将60只Wistar大鼠随机分成A组、B组、C组和D组四个组,每组15只.四组均行右坐骨神经损伤模型,术毕向B组、C组和D组坐骨神经内注入滴度分别为2＆#215;10-6 TU、2＆#215;10-7 TU、2＆#215;10-8 TU的Lenti-RFP悬液8μl,向A组注入8μl平衡盐液（BSS）作为对照组.术后第1、2、4周观察红色荧光蛋白的表达情况,计算转染效率;并进行神经组织学检查.结果 术后第1周B、C、D组均出现RFP表达,术后第4周D组的转染效率最高.同一时间不同滴度组以及同一滴度组不同时间比较差异有统计学意义（P＜0.05）.组织学检查病毒载体对神经未见明显炎性反应及组织学损伤.结论 慢病毒载体能安全有效的在体内转染雪旺细胞,并有浓度依赖性和时间依赖性,对组织无明显毒性作用.

Objective To investigate the expression of RFP in Schwann cell of rat sciatic nerve transfected with lentivirus vetor.Methods Sciatic nerve injury model was established by transection of right sciatic nerve in 60 female wistar rats that were then randomly divided into 4 groups：group A,B,C and D.At the end of the procedure,8μl of Lenti-RFP suspension （2× 10-6 TU,2 × 10-7 TU,2 × 10-8 TU） were injected into the sciatic nerve of the rats in group B,C and D,respectively.The rats in group A received 8μl of balanced salt solution （BSS） and were used as the control.In the 1st,2nd and 4th week after the surgery,the expression of RFP and the effects of lentivirus vetor on Schwann cells were tested,and the transfection efficiency was calculated.Results RFP gene expressions were observed in Group B,C and D one week after surgery,and the highest transfection efficiency were found in group D at the 4th week after surgery..Statistically significant differences of transduction efficiency were observed in the different dosage groups at the same time,or in the same group at different time.The histological damage to sciatic nerve caused by the vector was not obvious in every group.Conclusions RFP gene can be transfected into Schwann cells in vivo with stable and efficient expression.Clear dosage-dependent and time-dependent features were observed.Lentivirus vector caused no histological damage to sciatic nerve.