转基因中国仓鼠卵细胞中单胺囊泡转运体抗毒性作用的研究

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。

A Study on Antitoxic Role of Vesicular Monoamine Transporter 2 in Transgenic Chinese Hamster Ovary Cells

Objective : To study the antitoxic role of vesicular monoamine transporter 2 (VMAT2) in transgenic Chinese Hamster ovary (CHO) cell. Methods :With the technology of trans-gene from PC 12 to CHO, MTT reduction assay was used to detect MPP+ toxic effect on wild type CHO (wtCHO) and transgenic CHO. Meanwhile, the role of reserpine was also observed in MPP+ toxic effects. Results :The sensitivity of transgenic CHO to MPP+ was much less than that of wtCHO with 0. 5 mmol/L MPP+. Transgenic CHO had the same sensitivity as wtCHO if rotenone was given. WtCHO, by given reserpine alone, didn't change its sensitivity to MPP+. Conclusions :VMAT2 has protective effect on transgenic CHO by transporting MPP+ to vesicles.