摘要
目的探讨自噬是否参与老龄SD大鼠多巴胺(DA)能神经元对Rotenone神经毒性的易感性。方法♂SD大鼠,分别构建3、12、20mon组,每个月龄组又随机分为自然增龄组和Rotenone处理组,以Rotenone1.0mg·kg-1.d-1背部皮下注射,连续给药30d,每周停药1d,构建SD大鼠Rotenone染毒模型。大鼠黑质连续切片,进行酪氨酸羟化酶(tyrosine hydroxylase,TH)免疫组化染色;Western blot检测黑质部位微管相关蛋白轻链-3(LC3)蛋白表达水平;透射电镜观察黑质神经元中自噬体形成和溶酶体激活。结果在自然增龄组,大鼠黑质TH阳性细胞计数以12mon组为最高;Rotenone处理后各月龄组TH阳性细胞计数均比相应自然增龄组降低(P<0.01),且20mon组降低最为明显。Western blot结果显示,自然增龄组大鼠黑质部位LC3表达在20mon组比3mon组下降(P<0.01),Rotenone处理后各月龄组大鼠黑质部位LC3表达均比相应自然增龄组上调(P<0.01)。透射电镜显示Rotenone处理后大鼠黑质神经元可见自噬小体和溶酶体数目增多。结论自噬参与老龄大鼠DA能神经元对Rotenone神经毒性的易感性。
Aim To study whether autophagy participates in Rotenone-induced dopaminergic neurotoxicity in old SD rat. Method Rotenone neurotoxin SD rat model was constructed in different aged （3, 12, 20 months old,n = 20 in each group） rats. The Rotenone （ 1.0 mg·kg^-1·d^-1 , at 24 h intervals） was injected subcutaneously in back. Then, the tyrosine hydroxylase-immunoreactivity in substantia nigra pars compacta （SNc） was observed. Protein levels of LC3 were determined with Western Blot analysis. Autophagosomes were observed with electron microscopy. Result In normal aging groups, 12 months old rats had the highest number of SNc TH positive cells. There was a significant decrease of SNc TH positive cells in all Rotenone treated rats with the least number of 20 months old rats. Western blot analysis revealed the increase in LC3 protein levels after Rotenone treatment comparing with normal aging groups （P 〈0.01 ）, while the number of LC3 protein was least in 20 months old groups. Electron microscopy revealed that the formation of autophagosomes in SNc in Rotenone treated groups increased. Conclusion Activation of autophagy may be a prosurvival feedback response in Rotenone -induced dopaminergic neurotoxicity in old SD rats.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2009年第9期1157-1160,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30870869)
江苏省脑病生物信息重点实验室开放课题(NoJsbl0702)
关键词
帕金森病
老化
自噬
鱼藤酮
LC3
透射电镜
Parkinson＇ s disease
aging
autophagy
Rotenone
LC3
electron microscopy
