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[目的]研究三氯乙烯(TCE)染毒对L-02肝细胞一氧化氮(NO)合成和诱导型一氧化氮合酶(iNOS)表达水平的影响。[方法]用TCE不同浓度(0、0.125、0.25、0.5、1.0、2.0mmol/L)和1.0mmol/L TCE不同染毒时间(0、1、2、6、12、24h)染毒肝细胞。按试剂盒说明检测细胞培养液中NO浓度;提取肝细胞RNA,用荧光定量聚合酶链式反应(PCR)检测肝细胞iNOS mRNA相对表达量。用全自动生化分析仪测定TCE染毒肝细胞后培养液中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和乳酸脱氢酶(LDH)活性。[结果]不同浓度TCE染毒肝细胞24h后,各染毒组和对照组比较,NO和iNOS mRNA表达水平均明显升高;用同一浓度TCE(1.0mmol/L)染毒不同时间,≥2h染毒时间引起NO合成和iNOS mRNA表达水平升高。TCE各剂量组染毒后ALT、AST和LDH高于对照组。[结论]TCE染毒肝细胞可诱导肝细胞iNOS mRNA表达水平升高和NO合成增加,并对肝细胞有一定的损伤作用,提示TCE的肝细胞损害作用可能与NO产生有关。
[Objective] To investigate the effects of TCE on nitric oxide (NO) synthesis and inducible nitric oxide synthase (iNOS) expression in L-02 hepatocytes. [Method] The hepatocytes were exposed to different concentrations of TCE (0,0.125,0.25,0.5,1.0,2.0 mmol / L) and 1.0 mmol / L TCE for different exposure time (0, 1, 2, 6, 12, 24 h) . The concentrations of NO in the cell culture medium were detected by kit instructions. The RNA of hepatocytes was extracted and the relative expression of iNOS mRNA in liver cells was detected by fluorescence quantitative polymerase chain reaction (PCR). The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in the culture fluid of TCE-treated hepatocytes were measured by automatic biochemical analyzer. [Result] The mRNA expression levels of NO and iNOS were significantly increased in all exposure groups compared with the control group at different concentrations of TCE for 24 h. At the same concentration of TCE (1.0 mmol / L) for 2 h Exposure time caused NO synthesis and iNOS mRNA expression increased. The levels of ALT, AST and LDH in TCE groups were higher than those in control group. [Conclusion] TCE can induce the increase of iNOS mRNA expression and the increase of NO synthesis in hepatocytes, and have a certain damage on hepatocytes, suggesting that the hepatocellular damage induced by TCE may be related to the production of NO.