摘要
目的探讨施万细胞(Schwann cells,Scs)源性神经生长因子(nerve growth factor,NGF)及其受体(TrkA、P75)在丙烯酰胺(acrylamide,ACR)所致运动神经元损伤及损伤后自然修复过程中的变化。方法原代培养大鼠Scs,插入式混合培养Scs与运动神经元VSC4.1,ACR(终浓度为40μg/ml)染毒24 h,染毒后去除毒物暴露24 h,建立ACR损伤及损伤后自然修复的细胞模型;Western-blot和RT-PCR分别检测混合培养体系中Scs所分泌的NGF及其受体TrkA、P75的蛋白和mRNA水平变化。结果 ACR对混合培养体系内的运动神经元VSC4.1具有明显细胞毒性,细胞形态损伤显著,去除ACR暴露后损伤减轻;与对照组相比,ACR染毒24 h及自然修复24 h,NGF蛋白含量显著增加(P<0.05),ACR染毒24 h后P75含量减少(P<0.05),修复24 h,P75的含量变化不显著(P>0.05),未检出TrkA蛋白表达;与对照组相比,ACR染毒24 h,NGF mRNA、TrkA mRNA、P75 mRNA表达水平显著升高(P<0.05);修复24 h,NGF mRNA表达水平无明显改变(P>0.05),TrkA mRNA、P75 mRNA水平显著降低(P<0.05)。结论施万细胞源性NGF在ACR对运动神经元VSC4.1损伤及修复过程中与其受体结合,通过正负反馈调节发挥一定的神经元保护作用。
Objective To explore the changes of nerve growth factor and its receptors secreted by 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. Levels of protein and mRNA of NGF, TrkA and P75 were examined by Western-blot and RT-PCR. Results ACR has toxic effect on VSC4. 1 cells. Morphological changes can be found after treated with ACR for 24 hours and repair for 24 hours. As for groups treated with ACR （40 μg/ml） for 24 hours and repair for 24 hours, the protein level of NGF was increased compared with the control group （P 〈 0. 05）. As for groups treated with ACR （40 μg/ml） for 24 hours, the protein level of P75 was decreased compared with the control group （P 〈 0.05）. As for groups repair for 24 hours, the protein level of P75 had no significant difference with control group （P 〉0. 05）. No TrkA protein was found in this study. For group treated with ACR （40 μg/ml） for 24 hours, mRNA of NGF, TrkA and P75 was increased compared with the control group （P 〈 0. 05）. As for groups repair for 24 hours, mRNA of TrkA and P75 were increased compared with the control group （P 〈 0. 05）, but change of NGF mRNA was not significant. Conclusion NGF secreted by Schwan cells has beneficial effects through combination with its receptors on damage and repair of motor neuron induced by acrylamide.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2012年第6期407-411,共5页
Journal of Toxicology
基金
国家自然科学基金(30901219)
环保公益性行业科研专项(201109038)
