饮用水有机提取物对L02细胞增殖及细胞周期的影响
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摘要
目的:探讨饮用水有机提取物对人正常肝细胞(L02)的细胞增殖及细胞周期的影响。方法:体外培养L02细胞,将其分别暴露于培养液(空白对照组)、0.1%DMSO(溶剂对照组)和0.312 5、0.625 0、1.250 0、2.500 0、5.000 0 L/m L固相萃取法提取的饮用水有机提取物(染毒组)中24、48及72 h,采用MTT法观察L02细胞的增殖情况,流式细胞仪检测细胞周期的构成比。结果:(1)与溶剂对照组比较,各染毒组细胞活力随着有机提取物浓度的增加及时间的延长而降低,差异有统计学意义(P<0.05);(2)随染毒剂量的增加,细胞周期中G0/G1期细胞比例逐渐下降,除24 h、48 h 0.312 5 L/m L染毒组外,其余剂量染毒组G0/G1期细胞比例均明显低于溶剂对照组(P<0.05);S期细胞比例则随有机提取物浓度的增加而上升,除24、48及72 h 0.312 5 L/m L的染毒组外,其余染毒组中S期细胞比例的增加与对照组比较,差异有统计学意义(P<0.05);染毒48 h、72 h后,L02细胞中G0/G1期细胞比例低于24 h染毒组(P<0.05);S期细胞比例则高于24 h染毒组(P<0.05)。结论:在本研究条件下,饮用水有机提取物可以阻滞L02细胞于S期,抑制细胞的增殖,且呈时间、剂量依赖性。
Objective: To investigate the influence of organic extracts from drinking water on liver cell( L02) proliferation and cell cycle. Methods: The L02 cells was cultivated in vitro: blank control group( culture solution),solvent control group( 0. 1% DMSO); organic extracts group( The dose of pollutants groups were 0. 312 5,0. 625 0,1. 250 0,2. 500 0 and 5. 000 0 L / m L samples were extracted by solid phase extraction method). Each group was treated for 24 h,48 h and 72 h. Cell proliferation was measured by MTT assay. The constituent ratio of cell cycle was detected with flow cytometry.Results:( 1) Compared with solvent control group,cell vitality of L02 cells decreased gradually with the increasing of exposure time and concentration of organic extracts. The differences were statistically significant( P < 0. 05).( 2) The proportion of cells in G0 / G1 phase decreased gradually with the increasing concentration of organic extracts. Except for 0. 312 5 L / m L of pollutants group at 24 h and48 h,significant differences were found in other exposure groups( P < 0. 05). The proportion of cell in S phase increased gradually with the increasing concentration of organic extracts. Except for0. 312 5 L / m L at 24 h,48 h and 72 hr exposure,significant differences existed compared with solvent control group,differences were statistically significant( P < 0. 05). Compared with 24 h exposure group,exposed groups at 48 h- exposure and 72 h- exposure demonstrated significantly increase in G0 / G1 phase cell proportion and S phase cell proportion with increasing time( P < 0. 05). Conclusions: The results of the present paper showed that the organic extracts from drinking water can inhibit cell proliferation and change cell cycle of L02 cells.
Objective: To investigate the influence of organic extracts from drinking water on liver cell( L02) proliferation and cell cycle. Methods: The L02 cells was cultivated in vitro: blank control group( culture solution),solvent control group( 0. 1% DMSO); organic extracts group( The dose of pollutants groups were 0. 312 5,0. 625 0,1. 250 0,2. 500 0 and 5. 000 0 L / m L samples were extracted by solid phase extraction method). Each group was treated for 24 h,48 h and 72 h. Cell proliferation was measured by MTT assay. The constituent ratio of cell cycle was detected with flow cytometry.Results:( 1) Compared with solvent control group,cell vitality of L02 cells decreased gradually with the increasing of exposure time and concentration of organic extracts. The differences were statistically significant( P < 0. 05).( 2) The proportion of cells in G0 / G1 phase decreased gradually with the increasing concentration of organic extracts. Except for 0. 312 5 L / m L of pollutants group at 24 h and48 h,significant differences were found in other exposure groups( P < 0. 05). The proportion of cell in S phase increased gradually with the increasing concentration of organic extracts. Except for0. 312 5 L / m L at 24 h,48 h and 72 hr exposure,significant differences existed compared with solvent control group,differences were statistically significant( P < 0. 05). Compared with 24 h exposure group,exposed groups at 48 h- exposure and 72 h- exposure demonstrated significantly increase in G0 / G1 phase cell proportion and S phase cell proportion with increasing time( P < 0. 05). Conclusions: The results of the present paper showed that the organic extracts from drinking water can inhibit cell proliferation and change cell cycle of L02 cells.
引文
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[2]王文忠.浅析水环境中有毒污染物的来源和危害[J].山西水土保持科技,2013(3):30-31.
[3]韩方岸,将兆峰,柯士呜,等.镇江农村肿瘤高发的病因学调查[J].预防医学情报杂志,2011(1):1-4.
[4]高红萍,王丽,白钢,等.包头市黄河水与自来水有机提取物致突变性研究[J].包头医学院学报,2007(6):588-589.
[5]田怀军,吴德生,陶锐,等.氯化饮用水有机提取物对睾丸支持细胞的毒性效应[J].环境与健康杂志,2001(3):146-147.
[6]杨光红,王士然,张爱华,等.G市某区管网末梢水中有机提取物对雄性大鼠生殖激素分泌水平的影响[J].环境与职业医学,2013(5):342-345.
[7]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(9):799-801.
[8]徐文零,杨光红,王士然,等.G市某区管网末梢水有机污染物对大鼠精子形态的影响[J].贵阳医学院学报,2013(2):134-137.
[9]李骞,敖云霞,杨光红,等.饮用水有机提取物致大鼠肝脏DNA及蛋白质氧化损伤的研究[J].现代预防医学,2014(22):4138-4140.
[10]孟凡生,王业耀,陈晶.我国水环境有机物分析前处理技术[J].环境监测管理与技术,2010(4):15-18.
[11]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:华中科技大学博士研究生论文,2011.
[12]李春.细胞周期及其调控[J].福建医药杂志,2013(5):146-147.
[13]高世勇,曲笑莹.细胞周期同步化研究进展[J].中国药理学通报,2014(1):17-21.
[14]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(11):984-986.
[15]Xu L,Wang C,Wen Z,et,al.Selective up-regulation of CDK2 is critical for TLR9 signaling stimulated proliferation of human lung cancer cell[J].Immunol Lett,2010(2):93-99.