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目的研究三氯乙烯(TCE)对人肝细胞凋亡基因(Caspase-3、Caspase-8、BAX、Bcl-2)和肝代谢酶基因(CYP1A2、CYP2E1、CYP3A4)mRNA表达水平的影响。方法用含10%胎牛血清的RPMI-1640培养肝细胞,DMSO作为溶剂对照,采用不同浓度TCE(0.2、0.4、0.8、1.6、3.2mmol/L)染毒L02肝细胞24h,另外用TCE 1.0mmol/L染毒细胞不同时间(2、4、8、16、32h),采用Real-time荧光定量PCR技术检测肝细胞中凋亡基因和CYP基因mRNA表达水平。结果不同剂量TCE染毒后,凋亡基因Caspase-3、Caspase-8、BAX表达水平比对照组升高21%~145%,差异有统计学意义(P<0.05或P<0.01),Bcl-2表达水平下降20%~35%。TCE染毒引起CYP1A2、CYP3A4、CYP2E1的mRNA表达明显上升,比对照组升高30%~550%,差异有统计学意义(P<0.05或P<0.01)。用TCE 1.0mmol/L染毒2~32h,同样发现凋亡基因和肝代谢酶基因表达增加,与对照组比较差异有统计学意义(P<0.05或P<0.01)。结论 TCE可引起肝细胞凋亡基因和肝代谢酶基因表达变化,凋亡基因和肝代谢酶基因可能在TCE对肝细胞毒性效应中发挥重要作用。
Objective To investigate the effect of trichlorethylene (TCE) on the mRNA expression of apoptosis genes (Caspase-3, Caspase-8, BAX, Bcl-2) and hepatic metabolism genes (CYP1A2, CYP2E1, CYP3A4) in hepatocytes. Methods Liver cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum. DMSO was used as solvent control. Twenty-two L02 hepatocytes were exposed to TCE (0.2,0.4,0.8,1.6,3.2mmol / L) for 24 h. TCE 1.0 (2, 4, 8, 16, 32h). The mRNA expression of apoptosis gene and CYP gene in hepatocytes was detected by Real-time fluorescent quantitative PCR. Results Compared with control group, the expression of Caspase-3, Caspase-8 and BAX in TCE group increased by 21% -145%, the difference was statistically significant (P <0.05 or P <0.01) 2 expression decreased by 20% to 35%. TCE exposure induced a significant increase of mRNA expression of CYP1A2, CYP3A4 and CYP2E1, which was 30% ~ 550% higher than that of the control group (P <0.05 or P <0.01). With TCE 1.0 mmol / L for 2 ~ 32h, the gene expression of apoptosis gene and hepatic metabolism enzyme also increased, which was significantly different from the control group (P <0.05 or P <0.01). Conclusion TCE can induce the gene expression changes of hepatocyte apoptosis and liver metabolism, and the gene of apoptosis and liver metabolism may play an important role in the cytotoxic effect of TCE on hepatocytes.