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目的:研究转基因中国仓鼠卵(CHO)细胞中单胺囊泡转运体(VMAT_2)的抗毒性作用。方法:利用转PC12细胞基因到CHO细胞中形成的转基因CHO细胞(cDNA-CHO),采用MTT比色法检测1-甲基-4-苯基吡啶离子(MPP~+)对CHO细胞野生株(wtCHO)和cDNA-CHO细胞的毒性作用,并观察利血平——VMAT_2的特异性阻滞剂对MPP~+毒性作用的影响。结果:在MPP~+ 0.5mmol/L以上浓度cDNA-CHO细胞对MPP~+敏感性比wtCHO低得多;cDNA-CHO和wtCHO对鱼藤酮(rotenon)的敏感性无显著差异;加入利血平后,上述保护作用消失,cD-NA-CHO对MPP~+敏感性与wtCHO细胞无差异,而单独予以wtCHO细胞利血平则不能改变它对低浓度MPP~+的敏感性。结论:此保护机制是由转基因细胞中VMAT_2引起的,VMAT_2在转基因的非神经细胞系(CHO细胞系)中也能将MPP~+转运至囊泡内,从而保护细胞,同时也提示PC12细胞内具有抗毒性作用的成分。
Objective: To study the anti-toxicity effect of monoamine vesicular transporter (VMAT_2) in transgenic Chinese hamster ovary (CHO) cells. Methods: MTT assay was used to detect the effect of MPP ~ + on the growth of CHO cell line (CHO-1) by using transgenic CHO cells (CHO-CHO) wtCHO) and cDNA-CHO cells. The effects of reserpine-VMAT_2-specific blockers on MPP ~ + toxicity were also observed. Results: The sensitivity to MPP ~ + of cDNA-CHO cells was much lower than that of wtCHO at the concentration of MPP ~ + 0.5mmol / L. The sensitivity of rotenone to cDNA-CHO and wtCHO was not significantly different. After adding reserpine , The protective effect disappeared, cD-NA-CHO on the MPP ~ + sensitivity and wtCHO cells no difference, while alone wtCHO cells reserpine can not change its sensitivity to low concentrations of MPP ~ +. CONCLUSION: This mechanism of protection is caused by VMAT_2 in transgenic cells. VMAT_2 also transports MPP ~ + into vesicles in transgenic non-neuronal cell lines (CHO cell lines), thereby protecting cells. It also suggests that PC12 cells Anti-toxic effect of the ingredients.