摘要
目的探讨1,2-二氯乙烷(1,2-DCE)及其体内代谢产物对人星形胶质细胞(HAs)的毒性。方法取对数生长期的HAs建立离体细胞培养实验模型,分别采用不同剂量的1,2-DCE(5.00、10.00、25.00、50.00、100.00mmol/L)、2-氯乙醇(5.00、25.00、50.00、100.00、200.00 mmol/L)、2-氯乙醛(1.00、5.00、10.00、20.00、50.00mmol/L)和氯乙酸(0.01、0.05、0.10、0.50、1.00 mmol/L)对HAs进行染毒,培养24 h后,采用荧光倒置相差显微镜观察HAs形态,采用CCK-8比色法检测HAs的存活率和抑制率,估算24 h半抑制浓度(24 h-IC50),采用磷脂结合蛋白Ⅴ-异硫氰酸荧光素/碘化丙啶双标记流式细胞技术检测细胞的凋亡情况。结果染毒24 h后,1,2-DCE、2-氯乙醇、2-氯乙醛和氯乙酸均可使HAs发生不同程度的改变,表现为胞体变小、伪足变尖、胞内颗粒增多,细胞透明度下降,圆形悬浮细胞增多。随着1,2-DCE、2-氯乙醇、2-氯乙醛和氯乙酸染毒剂量的增加,HAs的细胞存活率均下降(P<0.01),细胞抑制率均增加(P<0.01),均呈剂量-效应关系;上述4种受试物对HAs的24 h-IC50分别为56.25、235.00、26.43和1.38 mmol/L。1,2-DCE、2-氯乙醇和氯乙酸均可诱导HAs凋亡,HAs的凋亡率与上述3种受试物的染毒剂量均呈正相关(P<0.01)。结论 1,2-DCE及其代谢产物2-氯乙醇、2-氯乙醛和氯乙酸均可对HAs造成毒性损伤,并可诱导HAs发生凋亡,以氯乙酸毒性作用最强。
Objective To explore the toxicity of 1,2-dichloroethane（ 1,2-DCE） and its metabolites on human astrocytes（ HAs）. Methods Different doses of 1,2-DCE（ 5. 00,10. 00,25. 00,50. 00 and 100. 00 mmol/L）,2-chlorohydrins（ 5. 00,25. 00,50. 00,100. 00 and 200. 00 mmol/L）,2-chloroacetaldehyde（ 1. 00,5. 00,10. 00,20. 00 and 50. 00 mmol / L） and chloroacetic acid（ 0. 01,0. 05,0. 10,0. 50 and 1. 00 mmol / L） were used for treating HAs in vitro during their logarithmic phase. After 24 hours of culture,the morphology of HAs was observed by fluorescent inverted phase contrast microscope. The survival rate and the inhibition ratio of HAs were detected by CCK-8 colorimetry to estimate the50% inhibiting concentration in 24 hours（ 24 h-IC50）. The apoptosis of HAs was tested by double-labeling and flow cytometry using Annexin Ⅴ-fluorescein isothiocyanate and propidium iodide. Results The morphology of HAs changed in varying degrees after 24 hours exposure to 1,2-DCE,2-chlorohydrins,2-chloroacetaldehyde and chloroacetic acid. The changes included smaller size of cells,pseudopodia tapering,increased intracellular particles and suspension of circular cells and decreased transparency of cells. With the increasing does of 1,2-DCE,2-chlorohydrins,2-chloroacetaldehyde and chloroacetic acid exposure,the survival rates of HAs decreased（ P〈0. 01）,while its inhibition ratios increased（ P〈0. 01）. They all showed dose-effect relationship. 24 h-IC50 of the above 4 chemicals were 56. 25,235. 00,26. 43 and1. 38 mmol / L,respectively. The 1,2-DCE,2-chlorohydrins and chloroacetic acid could induce the apoptosis of HAs and the apoptosis rate of HAs was positively correlated with the 3 kinds of chemicals（ P〈0. 01）. Conclusion 1,2-DCE and its metabolites 2-chloroacetaldehyde,2-chlorohydrins and chloroacetic acid can lead to toxic damage and induce the apoptosis of HAs. Chloroacetic acid has the strongest toxicity among the metabolites.
出处
《中国职业医学》
CAS
北大核心
2016年第1期30-36,共7页
China Occupational Medicine
基金
国家临床重点专科建设项目(2011-09)
广东省职业病防治重点实验室(2012A061400007)
广东省化学中毒与核辐射突发事件医学救援应急技术研究中心(2012A032100002)
广东省自然科学基金(S2013010011816)
广东省医学科学技术研究基金(B2014017,A2012068)
关键词
1
2-二氯乙烷
2-氯乙醇
2-氯乙醛
氯乙酸
人星形胶质细胞
细胞毒性
半抑制浓度
1
2-Dichloroethane
2-Chlorohydrins
2-chloroacetaldehyde
Chloroacetic acid
Human astrocyte
Cell toxicity
50% Inhibiting concentration
