雪旺细胞在ACR对运动神经元损伤及修复过程中的作用研究

Effects of Schwan cells on damage and repair of motor neuron induced by acrylamide

目的探讨雪旺细胞(Schwann cells,SCs)在丙烯酰胺(acrylamide,ACR)对运动神经元损伤及修复过程中的作用。方法原代培养大鼠SCs,用插入式培养皿进行SCs与运动神经元VSC4.1的混合培养;噻唑蓝(MTT)比色法检测不同浓度ACR对单独和混合培养运动神经元VSC4.1相对存活率的影响;细胞免疫荧光法检测ACR损伤和修复过程中运动神经元VSC4.1中生长相关蛋白43(growth-associated protein,GAP-43)水平的变化。结果 ACR染毒运动神经元VSC4.1 24 h,单独培养组的IC50为329μg/ml,混合培养的IC50为558μg/ml;GAP-43的水平较对照明显降低,混合培养组GAP-43的平均荧光强度(0.076±0.008)高于单独培养组(0.037±0.013),P<0.05。VSC4.1染毒24 h后去除ACR暴露并继续培养24 h,GAP-43的水平较染毒24 h时上升,混合培养组GAP-43的平均荧光强度值(0.241±0.033)高于单独培养组(0.143±0.034),P<0.05。结论 ACR对运动神经元VSC4.1具有明显的毒性作用,SCs对ACR所致运动神经损伤具有保护作用,同时对损伤后修复过程发挥一定的促进作用。

Objective To explore the effects of Schwan cells（SCs） on damage and repair of motor neuron induced by acrylamide.Methods Primary culture of rat SCs,subculture of VSC4.1 and co-culture of these two kinds of cells were conducted.Cell relative survival rate and changes of GAP-43 were examined by MTT and cellular immunofluorescence method.Results MTT tests showed that IC50 of VSC4.1 treated with acrylamide for 24 h was 329 μg/ml,and for the co-cultured was 558μg/ml.Compared with the non co-cultured VSC4.1,the relative survival rates of co-cultured VSC4.1 were increased in the treatment groups（P0.05）.As for the groups treated with ACR for 24 hours,the expression of GAP-43 in both co-cultured groups and non co-cultured groups were decreased compared with the control group.The expression of GAP-43 in co-cultured group was more than that in the non co-cultured group（P0.05）.As for groups repair 24 hours after treatment with ACR,the expression of GAP-43 in both co-cultured groups and non co-cultured groups was less than that in the control group but more than groups treated with ACR for 24 hours.The expression of GAP-43 in co-cultured group was more than that in the non co-cultured group（P0.05）.Conclusion Acrylamide has obvious toxic effect on motor neuron VSC4.1.SCs might have protective effects on damage induced by acrylamide.And Scs can promote the repair of motor neuron.