饮用水有机提取物对正常人肝细胞凋亡及炎性因子的影响
|
摘要
目的观察L02细胞经饮用水有机提取物染毒后所诱发的白介素6(interleukin 6,IL-6)和丙氨酸氨基转移酶(alanine transaminase,ALT)及细胞凋亡的改变。方法采用固相萃取法提取饮用水中有机污染物。将取处于对数生长期的正常人肝(L02)细胞分别暴露于分别暴露于含终浓度0.312 5、0.625 0、1.250 0、2.500 0、5.000 0 L/ml有机提取物的培养液中暴露24、48、72 h,同时设溶剂对照(含0.1%DMSO培养液)和空白对照(培养液)。检测细胞凋亡率及细胞培养液上清中IL-6的含量和ALT的活力。结果与溶剂对照组比较,各剂量饮用水有机提取物染毒不同时间L02细胞的凋亡率均较高,除0.312 5、0.625 0 L/ml染毒24 h和0.312 5 L/ml染毒48 h组外,差异均有统计学意义(P<0.05);各剂量饮用水有机提取物染毒不同时间L02细胞培养液上清中IL-6的含量均较高,差异均有统计学意义(P<0.05);各剂量饮用水有机提取物染毒不同时间L02细胞培养液上清中ALT的活力均较高,除0.312 5、0.625 0 L/ml染毒24 h组外,差异均有统计学意义(P<0.05)。且随着饮用水有机提取物染毒剂量的升高和染毒时间的延长,L02细胞的凋亡率及细胞培养液上清中IL-6含量和ALT活力均呈上升趋势。结论在本实验条件下,饮用水有机污染物可剂量依赖性及时间依赖性地诱导L02细胞炎性因子IL-6的分泌,引发炎症反应,诱导细胞凋亡,从而导致肝细胞损伤。
Objective To observe the effect of organic extracts from drinking water on interleukin 6(IL-6),alanine transaminase(ALT) and apoptosis in hepatocytes(L02 cells). Methods Organic pollutants in water samples were extracted by solid phase extraction method. L02 cell were divided into seven groups: blank control group(culture medium),solvent control group(0.1%DMSO) and organic extracts groups(the dose of extracts groups were 0.312 5,0.625 0,1.250 0,2.500 0 and 5.000 0 L/ml respectively). Each group was treated for 24 h, 48 h and 72 h respectively. The rate of cell apoptosis was detected with flow cytometry. ELISA was selected to determine the concentrations of IL-6 in the supernatant. The activity of ALT was detected with automatic biochemical analyzer. Results Compared with the solvent control group,the apoptosis rates in organic extracts groups at different time points were obviously higher except for 0.312 5, 0.625 0 L/ml at 24 h and 0.312 5 L/ml at 48 h(P<0.05); The IL-6 levels in organic extracts exposure group at different time points were obviously higher(P<0.05). The activities of ALT in organic extracts exposure group at different time points were obviously higher except for 0.312 5,0.625 0 L/ml at24 h(P<0.05). With the increasing of organic extracts level and exposure time,the apoptosis rates,the IL-6 levels and the ALT activities in organic extracts exposure groups were obviously higher than control group(P <0.05). Conclusion The organic extracts from drinking water can dose-dependently or time-dependently induce the secretion of IL-6 in L02 cells and induce hepatocytes apoptosis.
Objective To observe the effect of organic extracts from drinking water on interleukin 6(IL-6),alanine transaminase(ALT) and apoptosis in hepatocytes(L02 cells). Methods Organic pollutants in water samples were extracted by solid phase extraction method. L02 cell were divided into seven groups: blank control group(culture medium),solvent control group(0.1%DMSO) and organic extracts groups(the dose of extracts groups were 0.312 5,0.625 0,1.250 0,2.500 0 and 5.000 0 L/ml respectively). Each group was treated for 24 h, 48 h and 72 h respectively. The rate of cell apoptosis was detected with flow cytometry. ELISA was selected to determine the concentrations of IL-6 in the supernatant. The activity of ALT was detected with automatic biochemical analyzer. Results Compared with the solvent control group,the apoptosis rates in organic extracts groups at different time points were obviously higher except for 0.312 5, 0.625 0 L/ml at 24 h and 0.312 5 L/ml at 48 h(P<0.05); The IL-6 levels in organic extracts exposure group at different time points were obviously higher(P<0.05). The activities of ALT in organic extracts exposure group at different time points were obviously higher except for 0.312 5,0.625 0 L/ml at24 h(P<0.05). With the increasing of organic extracts level and exposure time,the apoptosis rates,the IL-6 levels and the ALT activities in organic extracts exposure groups were obviously higher than control group(P <0.05). Conclusion The organic extracts from drinking water can dose-dependently or time-dependently induce the secretion of IL-6 in L02 cells and induce hepatocytes apoptosis.
引文
[1]蒋金花.水体有机污染物对人体健康的影响[J].国外医学:卫生学分册,2003,30(6):321-325.
[2]杨光红,张爱华,王士然,等.常规处理工艺对地表水有机提取物遗传毒性的影响[J].环境与健康杂志,2013,30(9):799-801.
[3]李鹏,崔志鹏,刘胜学,等.J江某段有机污染物对大鼠肝肾毒性作用[J].中国公共卫生,2006,22(4):409-413.
[4]赵淑华,隋春生,王春华,等.饮用水有机提取物对正常人肝HL-7702细胞的毒性[J].环境与健康杂志,2011,28(11):984-986.
[5]杨光红,桂晓玲,岑延利,等.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[J].环境与健康杂志,2014,31(10):920-923.
[6]金伯泉,熊思东.医学免疫学[M].北京:卫生出版社,2008:60-61.
[7]张喜平,林谦.急性胰腺炎时炎症介质与细胞凋亡关系的研究进展[J].世界华人消化杂志,2005,23(13):2773-2777.
[8]高润平,齐晓艳.肝纤维化的发生机制与治疗进展[J].世界华人消化杂志,2006,23(14):2263-2269.
[9]曹波,任谦,崔志鸿,等.嘉陵江C市段源水中有机污染物定性研究[J].四川环境,2006,25(6):78-80.
[10]陈曦.P53非依赖性信号通路在邻苯二甲酸二(2-乙基已基)酯致体外培养人肝细胞毒性中的调控作用[D].武汉:华中科技大学,2011.
[11]魏红山,李定国,陆汉明.细胞凋亡的信息转导机制与调节[J].国外医学:分子生物学分册,2000,22(1):27-31.
[12]Lawson JA,Fisher MA,Simmons CA,et al.Parenchymal cell apoptosis as a signal for sinusoidal sequestration and transendothelial migration of neutrophils in murine models of endotoxin and Fasantibody-induced liver injury[J].Hepatology,1998,28:761-767.
[2]杨光红,张爱华,王士然,等.常规处理工艺对地表水有机提取物遗传毒性的影响[J].环境与健康杂志,2013,30(9):799-801.
[3]李鹏,崔志鹏,刘胜学,等.J江某段有机污染物对大鼠肝肾毒性作用[J].中国公共卫生,2006,22(4):409-413.
[4]赵淑华,隋春生,王春华,等.饮用水有机提取物对正常人肝HL-7702细胞的毒性[J].环境与健康杂志,2011,28(11):984-986.
[5]杨光红,桂晓玲,岑延利,等.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[J].环境与健康杂志,2014,31(10):920-923.
[6]金伯泉,熊思东.医学免疫学[M].北京:卫生出版社,2008:60-61.
[7]张喜平,林谦.急性胰腺炎时炎症介质与细胞凋亡关系的研究进展[J].世界华人消化杂志,2005,23(13):2773-2777.
[8]高润平,齐晓艳.肝纤维化的发生机制与治疗进展[J].世界华人消化杂志,2006,23(14):2263-2269.
[9]曹波,任谦,崔志鸿,等.嘉陵江C市段源水中有机污染物定性研究[J].四川环境,2006,25(6):78-80.
[10]陈曦.P53非依赖性信号通路在邻苯二甲酸二(2-乙基已基)酯致体外培养人肝细胞毒性中的调控作用[D].武汉:华中科技大学,2011.
[11]魏红山,李定国,陆汉明.细胞凋亡的信息转导机制与调节[J].国外医学:分子生物学分册,2000,22(1):27-31.
[12]Lawson JA,Fisher MA,Simmons CA,et al.Parenchymal cell apoptosis as a signal for sinusoidal sequestration and transendothelial migration of neutrophils in murine models of endotoxin and Fasantibody-induced liver injury[J].Hepatology,1998,28:761-767.