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目的:探讨锰对PC12细胞蛋白酶体20s复合物活性的影响。方法:以MTT以及平板克隆形成实验筛选作用浓度,不同时间以及不同浓度作用之后,提取细胞总蛋白,分别检测总蛋白提取液中20s复合物3种酶的活性。结果:含有MnCl2300μmol/L的培养基对蛋白酶体3种酶活性的抑制率随着作用时间的延长而增高。作用3 d后,对3种酶活性的抑制率均超过20%。在作用时间相同的条件下,随着作用浓度的增高,锰对PC12细胞蛋白酶体活性的抑制作用逐渐增强。同样作用3 d后,当MnCl2浓度达到500μmol/L时,锰对3种酶活性的抑制率均超过40%。结论:蛋白酶体20s活性的抑制可能在锰所诱导的神经毒性过程中发挥着一定的作用。
Objective: To investigate the effect of manganese on the activity of proteasome 20s in PC12 cells. Methods: MTT assay and plate clone formation assay were used to screen the concentration of total protein, and the total protein was extracted at different time and concentration. The activities of three enzymes of 20s complex in total protein extract were determined. Results: The inhibitory rate of three enzyme activities of proteasome with MnCl2300μmol / L medium increased with time. After 3 days, the inhibitory rates of all three enzymes exceeded 20%. Under the same action time, the inhibitory effect of manganese on the proteasome activity of PC12 cells gradually increased with the increase of the action concentration. After the same effect for 3 d, the inhibitory rate of Mn to all three enzymes exceeded 40% when the MnCl2 concentration reached 500 μmol / L. Conclusion: Inhibition of 20s proteasome activity may play a role in the neurotoxicity induced by manganese.