二氯乙烷及氯乙酸对细胞周期及凋亡的影响研究

摘要/Abstract

摘要： 目的初步探索1,2-二氯乙烷和氯乙酸对大鼠神经胶质瘤细胞和大鼠雪旺细胞周期和凋亡的影响。方法 0、15、30、60 mmol/L 1,2-二氯乙烷和0、225、450、900 μmol/L氯乙酸分别与大鼠神经胶质瘤细胞和大鼠雪旺细胞共培养24 h,通过流式细胞仪检测细胞周期及凋亡的变化,评价1,2-二氯乙烷和氯乙酸对两种细胞的毒性作用。结果大鼠神经胶质瘤细胞和大鼠雪旺细胞的相对存活率均随着1,2-二氯乙烷和氯乙酸染毒剂量的增加而逐渐降低,1,2-DCE对大鼠神经胶质瘤细胞和大鼠雪旺细胞的IC₅₀值分别为（77.1±1.4）和（72.9±4.3）mmol/L,氯乙酸对大鼠神经胶质瘤细胞和大鼠雪旺细胞的IC₅₀值分别为（895.0±44.5）和（803.2±37.3）μmol/L。0、15、30、60 mmol/L 1,2-二氯乙烷对大鼠神经胶质瘤细胞凋亡率分别为（5.55±0.78）%、（4.93±0.08）%、（6.31±0.53）%和（38.93±2.53）%,G₀/G₁期细胞比值分别为（61.46±0.79）%、（62.43±2.13）%、（60.19±6.41）%和（43.39±4.82）%,S期细胞比值分别为（13.46±4.57）%、（16.89±3.25）%、（18.56±4.68）%和（32.23±3.49）%,G₂/M期细胞比值分别为（23.36±4.53）%、（18.58±5.57）%、（20.14±6.40）%和（23.12±6.46）%。60 mmol/L 1,2-二氯乙烷可引起大鼠神经胶质瘤细胞的凋亡率升高、G₀/G₁期细胞比值降低、S期细胞比值升高（均P<0.01）。0、15、30、60 mmol/L 1,2-二氯乙烷对大鼠雪旺细胞凋亡率分别为（5.50±1.89）%、（7.95±2.38）%、（7.89±0.99）%和（29.20±7.53）%,G₀/G₁期细胞比值分别为（39.08±3.20）%、（44.00±20.65）%、（42.15±18.03）%和（37.65±6.39）%,S期细胞比值分别为（28.15±3.62）%、（25.77±15.47）%、（25.20±7.58）%和（23.85±2.00）%,G₂/M期细胞比值分别为（32.02±2.78）%、（29.58±7.06）%、（31.49±12.27）%和（37.10±7.68）%。60 mmol/L 1,2-二氯乙烷可引起大鼠雪旺细胞凋亡率升高（P<0.01）。0、225、450、900 μmol/L氯乙酸对大鼠神经胶质瘤细胞凋亡率分别为（5.55±0.78）%、（6.87±4.07）%、（5.16±0.95）%和（5.70±1.21）%,G₀/G₁期细胞比值分别为（61.46±0.79）%、（60.15±7.92）%、（61.27±3.11）%和（63.57±3.51）%,S期细胞比值分别为（13.46±4.57）%、（18.81±5.95）%、（16.92±4.68）%和（13.99±1.34）%,G₂/M期细胞比值分别为（23.36±4.53）%、（18.93±1.96）%、（19.90±4.91）%和（20.86±2.03）%。0、225、450、900 μmol/L氯乙酸对大鼠雪旺细胞凋亡率分别为（5.50±1.89）%、（7.60±2.60）%、（9.20±2.92）%和（37.88±6.99）%,G₀/G₁期细胞比值分别为（39.08±3.20）%、（39.88±6.06）%、（37.30±9.58）%和（36.59±7.16）%,S期细胞比值分别为（28.15±3.62）%、（20.91±7.76）%、（22.01±7.28）%和（16.72±5.49）%,G₂/M期细胞比值分别为（32.02±2.78）%、（38.54±2.71）%、（38.74±5.21）%和（45.49±5.70）%。900 μmol/L氯乙酸可引起大鼠雪旺细胞凋亡率和G₂/M期升高（均P<0.05）。结论 1,2-二氯乙烷可引起大鼠神经胶质瘤细胞和大鼠雪旺细胞凋亡率升高、大鼠神经胶质瘤细胞周期阻滞。氯乙酸可引起大鼠雪旺细胞凋亡率升高、细胞周期阻滞。

关键词: 1,2-二氯乙烷, 氯乙酸, 大鼠神经胶质瘤细胞, 大鼠雪旺细胞, 细胞周期, 细胞凋亡

Abstract: Objective To preliminarily explore the effects of 1,2-dichloroethane and chloroacetic acid on the cell cycle and apoptosis in rat glioma cells and rat Schwann cells. Methods 0,15,30,60 mmol/L 1,2-dichloroethane and 0,225,450,900 μmol/L chloroacetic acid were co-cultured with rat glioma cells and rat Schwann cells for 24 h,respectively. The changes in the cell cycle and apoptosis were detected by flow cytometry,to evaluate the toxicity of 1,2-dichloroethane and chloroacetic acid in the two cells. Results With the increasing toxic doses of 1,2-dichloroethane and chloroacetic acid,the relative cell survival rate was gradually decreased in both cells. The IC₅₀ value of 1,2-dichloroethane was （77.1±1.4） and （72.9±4.3）mmol/L in rat glioma cells and rat Schwann cells respectively. The IC₅₀ value of chloroacetic acid was （895.0±44.5） and （803.2±37.3）μmol/L in rat glioma cells and rat Schwann cells respectively. The apoptosis rates of 0,15,30,60 mmol/L 1,2-dichloroethane in rat glioma cells were respectively （5.55±0.78）%,（4.93±0.08）%,（6.31±0.53）% and（38.93±2.53）%,the cell ratios in G₀/G₁ phase were respectively （61.46±0.79）%,（62.43±2.13）%,（60.19±6.41）% and（43.39±4.82）%,in S phase were respectively（13.46±4.57）%,（16.89±3.25）%,（18.56±4.68）% and（32.23±3.49）%,in G₂/M phase were respectively（23.36±4.53）%,（18.58±5.57）%,（20.14±6.40）% and（23.12±6.46）%. 1,2-dichloroethane can cause an increased rate of apoptosis,lower cell ratio in G0/G1 phase and increased cell ratio in S phase in rat glioma cells at the dose of 60 mmol/L（all P<0.01）. The apoptosis rates of 0,15,30,60 mmol/L 1,2-dichloroethane in rat Schwann cells were respectively （5.50±1.89）%,（7.95±2.38）%,（7.89±0.99）% and （29.20±7.53）%,and the cell ratios in G₀/G₁ phase were respectively （39.08±3.20）%,（44.00±20.65）%,（42.15±18.03）% and（37.65±6.39）%,in S phase were respectively（28.15±3.62）%,（25.77±15.47）%,（25.20±7.58）% and（23.85±2.00）%,in G₂/M phase were respectively （32.02±2.78）%,（29.58±7.06）%,（31.49±12.27）% and（37.10±7.68）%. 60 mmol/L 1,2-dichloroethane can cause an increased rate of apoptosis in rat Schwann cells（P<0.01）. The apoptosis rates of 0,225,450,900 μmol/L chloroacetic acid in rat glioma cells were respectively （5.55±0.78）%,（6.87±4.07）%,（5.16±0.95）% and（5.70±1.21）%,and the cell ratios in G0/G1 phase were respectively （61.46±0.79）%,（60.15±7.92）%,（61.27±3.11）% and （63.57±3.51）%,in S phase were respectively （13.46±4.57）%,（18.81±5.95）%,（16.92±4.68）% and （13.99±1.34）%,in G₂/M phase were respectively （23.36±4.53）%,（18.93±1.96）%,（19.90±4.91）% and （20.86±2.03）%. The apoptosis rates of 0,225,450,900 μmol/L chloroacetic acid in rat Schwann cells were respectively（5.50±1.89）%,（7.60±2.60）%,（9.20±2.92）% and （37.88±6.99）%,and the cell ratios in G₀/G₁ phase were respectively（39.08±3.20）%,（39.88±6.06）%,（37.30±9.58）% and（36.59±7.16）%,in S phase were respectively（28.15±3.62）%,（20.91±7.76）%,（22.01±7.28）% and（16.72±5.49）%,in G₂/M phase were respectively （32.02±2.78）%,（38.54±2.71）%,（38.74±5.21）% and（45.49±5.70）%. 900 μmol/L chloroacetic acid can cause an increased rate of apoptosis and increased cell ratio in G₂/M phase in rat Schwann cells（both P<0.05）. Conclusion 1,2-dichloroethane can cause increased apoptotic rate of rat glioma cells and Schwann cells,cycle arrest in rat glioma cells. Chloroacetic acid can cause increased apoptotic rate and cycle arrest in rat Schwann cells.

Key words: 1,2-dichloroethane, Chloroacetic acid, Rat glioma cells, Rat Schwann cells, Cell cycle, Cell apoptosis

中图分类号:

R114

曾丽海, 李宏玲, 谢植伟, 郑杰蔚, 李国樑, 殷霄. 二氯乙烷及氯乙酸对细胞周期及凋亡的影响研究. [J]职业与健康, 2023, 39(4): 433-439.

ZENG Li-hai, LI Hong-ling, XIE Zhi-wei, ZHENG Jie-wei, LI Guo-liang, YIN Xiao. Study on effects of dichloroethane and chloroacetic acid on cell cycle and apoptosis. [J]OCCUPATION AND HEALTH, 2023, 39(4): 433-439.

参考文献

[1] 张寿林.职业病防治知识讲座(十三) 急性职业性有机溶剂中毒(二)—二氯乙烷、正己烷[J].劳动保护,2002(11):52-53.
[2] 夏元洵.化学物质毒性全书[M].上海:上海科学技术出版社,1991:307.
[3] 高岚岳,齐莹,金亚平.1,2-二氯乙烷的神经毒性[J].中国工业医学杂志,2012,25(1):42-43,57.
[4] CHEN S,ZHANG Z,LIN H,et al.1,2-Dichloroethane-induced toxic encephalopathy:A case series with morphological investigations[J].Neurol Sci,2015,351:36-40.
[5] 李红梅,孙即彬,李侠,等.1例职业性急性中毒性脑病引起的思考[J].职业卫生与应急救援,2021,39(6):710-712.
[6] 王凌霄,罗超红,王高阳,等.ERK/AP-1信号通路在亚急性1,2-二氯乙烷中毒性脑水肿中的作用[J].中国工业医学杂志,2021,34(5):399-403+410,481.
[7] 胡函文,刘泰伶,陈育全,等.22例1,2-二氯乙烷中毒性脑病中内皮微颗粒的变化及临床意义[J].黑龙江医药,2020,33(2):275-278.
[8] 殷霄,贾肖辉,程浩,等.1,2-二氯乙烷对SD大鼠亚急性吸入毒性研究[J].中国职业医学,2015,42(4):378-384,391.
[9] 贾肖辉,程浩,徐丹丹,等.水通道蛋白4在1,2-二氯乙烷中毒性脑水肿中表达研究[J].中国职业医学,2016,43(2):138-142.
[10] WANG G,QI Y,GAO L,et al.Effects of subacute exposure to 1,2-dichloroethane on mouse behavior and the related mechanisms[J].Hum ExpToxicol,2013,32(9):983-991.
[11] 齐莹,石磊,高岚岳,等.亚急性1,2-二氯乙烷染毒对小鼠行为及脑神经递质含量的影响[J].中华劳动卫生职业病杂志,2011,19(6):413-416.
[12] 齐莹,石磊,高岚岳,等.1,2-二氯乙烷染毒对小鼠脑组织氨基酸类神经递质的影响[J].中国工业医学杂志,2010,23(6):439-441.
[13] 李国樑,郑杰蔚,殷霄,等.1,2-二氯乙烷亚急性暴露对大鼠抑郁样行为及单胺神经递质水平的影响[J].中国职业医学,2019,46(2):152-156.
[14] 李坤阳.低浓度1,2-二氯乙烷亚慢性暴露对小鼠海马谷氨酸代谢的影响及其机制研究[D].沈阳:中国医科大学,2020.
[15] 赵凤红,徐天胜,朴杰,等.亚慢性1,2-二氯乙烷染毒对小鼠空间学习记忆能力的影响[J].中国工业医学杂志,2018,31(4):276-278.
[16] BOWLER R M,GYSENS S,HARTNEY C.Neuropsychological effects of ethylene dichloride exposure[J].Neurotoxicology,2003,24(4-5):553-562.
[17] 陈育全,林毓嫱,蒋文中,等.职业性亚急性1,2-二氯乙烷中毒致精神行为异常13例[J].中国工业医学杂志,2018,31(6):432-433,448.
[18] YUAN L,YU W M,QU C K.DNA damage-induced G2/M chechpoint in SV40 large Tantigen-immortalized embryonic fibroblast cells requires SHP-2 tyrosine phosphatase[J].Biol Chem,2003,278(44):42812-42820.
[19] TAFAZOLI M,BAAETEN A,GEERLINGS P,et al.In vitro mutagenicity and genotoxicity study of a number of short-chain chlorinated hydrocarbons using the micronucleus test and the alkaline single cell gel electrophoresis technique(Comet assay) in human lymphocytes:a structure-activity relationship(QSAR) analysis of the genotoxic and cytotoxic potential[J].Mutagenesis,1998,13(2):115-126.
[20] CHEN Q,KANG J,FU C.The independence of and associations among apoptosis,autophagy,and necrosis[J].Signal Transduct Target Ther,2018(3):18.
[21] ZHANG Y,LI G,ZHONG Y,et al.1,2-Dichloroethane induce reproductive toxicity mediated by CREB/CREM signaling pathway in male NIH Swiss mouse[J].Toxicol Sci,2017,160(2):299-314.
[22] 陈铿铿,赖关朝,曾丽海,等.1,2-二氯乙烷通过线粒体通路诱导人星形胶质细胞凋亡研究[J].中国职业医学,2018,45(4):417-423.
[23] 李思,李宏玲,赵娜,等.1,2-二氯乙烷诱导SH-SY5Y细胞凋亡及对P53、BCL-2、BAX蛋白表达影响[J].中国职业医学,2017,44(2):164-169,175.
[24] 李晨.1,2-二氯乙烷染毒对体外培养SW620细胞增殖、周期和凋亡的影响研究[D].杭州:浙江大学,2012.
[25] 赖关朝,罗国忠,曾丽海,等.1,2-二氯乙烷及其代谢产物对人星形胶质细胞代谢研究[J].中国职业医学,2016,43(1):30-36.