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目的实验以PC12细胞系为实验对象进行研究,探讨丙烯醛对其毒性作用及其特征,为丙烯醛对AD的发病机制研究提供依据。方法通过细胞急性毒性实验测得其LC50;测定不同浓度丙烯醛处理后的细胞内总GSH含量和caspase 3酶活性,并通过western blot检测caspase3蛋白质在不同浓度丙烯醛处理后的表达情况。结果经CCK-8毒性实验和集落形成实验测定丙烯醛LC50分别为29.91μM、21.94μM;不同浓度丙烯醛处理24 h后可引起细胞内总GSH的下降;不同浓度丙烯醛处理24 h后细胞内caspase3酶活性未见明显上调,但短时间处理(2 h、4 h和8 h)可见明显上调倾向;western blot实验可见不同浓度丙烯醛处理8 h后有caspase3蛋白表达上调,而处理24 h后caspase3蛋白表达下降。结论丙烯醛对PC12细胞有较强的急性毒作用,其作用机制可能与丙烯醛诱导的细胞内GSH的耗竭和细胞凋亡以及细胞坏死有关。
Objective To study the PC12 cell line as an experimental object to investigate the toxic effects of acrolein and its characteristics, provide the basis for the study of acrolein on the pathogenesis of AD. Methods The LC50 of the cells was determined by acute toxicity test. The contents of total GSH and caspase 3 in cells treated with different concentrations of acrolein were determined. The expression of caspase 3 protein in different concentrations of acrolein was detected by western blot. Results The LC50 of acrolein was 29.91μM and 21.94μM, respectively, after CCK-8 toxicity test and colony formation assay. After treated with different concentrations of acrolein for 24 h, the total GSH levels were decreased. After treated with different concentrations of acrolein for 24 h, caspase3 activity was not significantly upregulated, but short-term treatment (2 h, 4 h and 8 h) showed a significant upward trend; Western blot experiment showed that the different concentrations of acrolein after 8 h caspase3 protein expression increased, but after 24 h Caspase3 protein expression decreased. Conclusions Acrolein has a strong acute toxicity on PC12 cells, and its mechanism may be related to acne-induced depletion of intracellular GSH and apoptosis and cell necrosis.