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目的探讨CDNF对6-羟多巴胺(6-OHDA)诱导的细胞凋亡效应的抑制作用。方法应用Li-po2000将重组真核表达质粒pDsRed-C1-CDNF转染至BMSCs,经G418筛选后获得稳定表达CDNF的MSCs细胞株(重命名为CDNF-MSCs细胞株)。应用MTT法和AnnexinⅤ-FITC细胞凋亡分析法分别检测CDNF-MSCs细胞上清液对6-OHDA诱导的PC12细胞凋亡作用的影响。结果通过G418筛选成功获得稳定表达CDNF的MSCs转染细胞,CDNF-MSCs细胞上清液预处理的PC12细胞对6-OHDA诱导的凋亡效应具有抑制作用,能有效的降低细胞的凋亡率。结论 CDNF作为一种分泌性蛋白,可显著抑制6-OHDA诱导PC12细胞凋亡。
Objective To investigate the inhibitory effect of CDNF on 6-hydroxydopamine (6-OHDA) -induced apoptosis. Methods Recombinant eukaryotic expression plasmid pDsRed-C1-CDNF was transfected into BMSCs by Li-po2000. MSCs stably expressing CDNF (renamed as CDNF-MSCs) were obtained by G418 screening. The effects of CDNF-MSCs supernatant on the apoptosis of PC12 cells induced by 6-OHDA were detected by MTT assay and AnnexinⅤ-FITC apoptosis assay. Results The MSCs transfected with CDNF stably transfected cells were successfully obtained by G418 screening. PC12 cells pretreated with CDNF-MSCs supernatant could inhibit the apoptosis induced by 6-OHDA and could effectively reduce the apoptosis rate. Conclusion As a secreted protein, CDNF can significantly inhibit 6-OHDA-induced PC12 cell apoptosis.