氯化消毒饮用水有机提取物对L-02肝细胞凋亡相关基因及蛋白表达的影响

Expression of apoptosis-related genes and proteins in L-02 cell exposed to organic extracts from chlorinated drinking water

目的探讨氯化消毒饮用水水样中有机提取物染毒对人肝细胞株(L-02)细胞凋亡相关基因及蛋白表达的影响。方法于2012年7—9月(丰水期)采集贵阳市某地水厂的管网末梢水水样并制备有机提取物。将L-02细胞分别于含0(对照)、0.312 5、0.625、1.25、2.5、5 L/ml氯化消毒饮用水有机提取物的培养基中暴露48 h。测定细胞中半胱氨酸蛋白酶3(caspase-3)、聚腺苷二磷酸核糖聚合酶1(PARP-1)、B细胞淋巴瘤2(Bcl-2)与B细胞淋巴瘤2相关x基因(Bax)m RNA及蛋白的表达水平。结果与对照组比较,2.5、5 L/ml饮用水有机提取物染毒组L-02细胞中caspase-3、Bax的m RNA及蛋白表达水平均较高,而0.625、1.25、2.5、5 L/ml饮用水有机提取物染毒组L-02细胞中Bcl-2、PARP-1的m RNA及蛋白表达水平均较低(除0.625 L/ml组PARP-1蛋白外),差异均有统计学意义(P<0.05)。结论在本实验条件下,较高剂量的氯化消毒饮用水有机提取物暴露可上调L-02肝细胞中caspase-3与Bax基因,下调Bcl-2及PARP-1基因,促进细胞凋亡的发生。

Objective To observe the effects organic extracts from chlorinated drinking water on the expression of apoptosis-related genes and proteins in L-02 cells. Methods The organic pollutants in the pipeline water were extracted by solid-phase extraction method. L-02 cell were exposed to organic extracts from chlorinated drinking water at the dose of 0（control）, 0.312 5,0.625, 1.25, 2.5 and 5 L/ml respectively. The exposure time lasted 48 hours. m RNA and protein expression levels of caspase-3,PARP-1,Bcl-2 and Bax were determined. Results Compared with the control group, m RNA and protein expression level of caspase-3 and Bax in 2.5,5 L/ml groups were up-regulated significantly（P〈0.05）, while m RNA and protein expression level of Bcl-2 and PARP-1 were significantly down-regulated in 0.625,1.25,2.5 and 5 L/ml groups（except for PARP-1 protein in 0.625 L/ml group）（P〈0.05）. Conclusion Under the condition of this experiment,exposing to higher level organic extracts from the chlorinated drinking water can up-regulate the expression of caspase-3 and Bax genes, and down-regulate the expression of Bcl-2 and PARP-1 genes.