CDNF在多巴胺能神经元变性中对自噬调控的研究

The effect of autophagy regulated by CDNF in degeneration of dopaminergic neurons

目的探讨CDNF对多巴胺能神经元变性的作用及其对自噬的调控。方法通过蛋白酶体抑制剂Lactacysin诱导PC12细胞变性前和后分别施加200nM的脑源性神经营养因子(cerebral dopamine neurotrophic factor,CDNF),利用MTT法检测PC12细胞生存率、RT-PCR和Western-blotting检测alpha-共核蛋白(synu-clein)mRNA和蛋白表达;利用Western-blotting检测p62、Beclin-1和LCI/II蛋白表达。结果经Lactacystin诱导后PC12细胞生存率为0.374±0.086、α-共核蛋白mRNA和蛋白表达分别为5.766±0.052和0.434±0.077,诱导前/后施加200nM的CDNF后PC12细胞生存率分别上升至(0.594±0.121和0.542±0.097)、α-共核蛋白mRNA和蛋白表达分别下降为(0.275±0.047、0.242±0.087和0.325±0.086、0.267±0.075);Beclin-1和p62分别下降为(0.728±0.143、0.235±0.096和0423±0.108、0.258±0.065),而LC3-I/II分别上升为(0.638±0.097和0.601±0.085)。结论CDNF可能通过负向调控自噬对多巴胺能神经元发挥神经保护和逆转作用。

Objective To investigate the effect of CDNF on degeneration of dopaminergic neu-rons and its regulation on autophagy.Methods PC12cells were treated with 200nM cerebral dopamine neurotrophic factor(CDNF)before and after denaturation.MTT assay The activity of PC12cells were detected by MTT assay,RT-PCR and Western blotting were used to detect the mRNA and protein ex-pression of alpha-synuclein was detected by Western blot,and the protein expressions of p62,beclin-1 and LCI/II were detected by Western blotting.Results After Lactacystininduction,the activity of PC12 cells decreased to 0.374±0.086and the mRNA and protein expression ofα-synuclein were 5.766±0.052and 0.434±0.077,respectively.After 200nM CDNF was applied before and after induction,the ac-tivity of PC12cells increased to 0.594±0.121and 0.542±0.097respectively,the mRNA and protein expression ofα-synuclein were decreased to 0.275±0.047、0.242±0.087and 0.325±0.086、0.267±0.075,beclin-1and p62decreased to 0.728±0.143、0.235±0.096and 0423±0.108、0.258±0.065re-spectively,while LC3-I/II increased to 0.638±0.097and 0.601±0.085respectively.Conclusion CD-NF may exert neuroprotective and reversal effects on dopaminergic neurons through negative regulating the autophagy.