二硫化碳对植入期小鼠子宫内膜细胞DNA损伤的研究

目的 用单细胞凝胶电泳(SCGE)技术检测二硫化碳(carbon disulfide,CS:)致植入期小鼠子宫内膜细胞DNA损伤,探讨其胚胎植人障碍机制.方法 用机械刮取法制备胚胎植入期小鼠子宫内膜单细胞悬液,台盼蓝检测细胞活性,用0、500、1000、2500μmol/L CS_2与细胞共培养1h后进行SCGE实验,采集彗旱图像,CASP系统自动分析各项指标.结果 不同剂量的CS_2染毒对DNA造成不同程度的损伤,形成较为典型的正常细胞和彗星细胞图像.与对照组比较,500、1000、2500 μmol/LCS_2染毒组彗星头部DNA含量百分比(HDNA%)分别减少了7.49%、12.19%、24.36%,差异均有统计学意义(P<0.01);500、1000、2500 μmol/LCS_2染毒组彗星尾部DNA含量百分比(TDNA%)、尾长(TL)、Olive尾矩(0TM)分别增加了7.13、11.60、23.18倍,3.68、5.98、9.62倍和9.16、16.84、39.32倍,差异均有统计学意义(P<0.01).与500、1000 μmol/L CS_2染毒组比较,2500μml/L CS_2染毒组的TDNA%、TL、彗星全长(CL)、尾矩(TM)、OTM增加了1.98、0.92、1.27、0.52、0.37倍和0.17、5.31、1.90、2.97、1.26倍,差异有统计学意义(P<0.01);与500 μmol/L CS_2染毒组比较,1000μmol/L CS_2染毒组的TDNA%、,TL、CL、TM、OTM增加了0.55、0.49、0.16、1.18、0.76倍,差异有统计学意义(P<0.01).HDNA%、TDNA%、TL、TM、OTM与染毒剂量的回归系数分别为-13.78,13.78,0.05,4.38,3.23,差异有统计学意义(P(0.01).结论 CS_2致植入期小鼠子宫内膜细胞DNA损伤,随着染毒剂量增加,损伤程度明显增加.CS_2损伤植入期小鼠母体子宫内膜细胞可能是影响胚胎正常植入的重要原因之一.

DNA damage of mice endometrial cells at implantation phase exposed to carbon disulfide

Objective To detect the DNA damage of mice endometrial cells induced by carbon disulfide with single cell gel electrophoresis(SCGE)and explore the mechanism of embryo implantation dis-order,Methods Endometrial cells,obtained by mechanical scrape,were used to test cell viability by trypan blue,Single cell suspension was exposed to the different concentrations of carbon disulfide(CS_2)at four dose groups(O,500,1000,2500 μmol/L).DNA damage wag detected by SCGE.The SCGE resuits were analyzed by CASP software.Results Different dosages of CS_2 concentration induced different varying degrees of damage,lorming typical normal cell and comet cell images.Compared with the control group,HDNA%decreased by 7.49%,12.1 9%and 24.36%respectively,and TDNA%,TL,OTM increased by 7.13,11.60,23.18,3.68.5.98.9.62,and 9.16,16.84,39.32 times respectively,in the groups of 500,1000,2500μmol/L CS_2(P<0.01).Com-pared with the group of 500,1000 μmol/L CS_2,TDNA%,TL CL,TM,OTM in the group of 2500μmol/L CS_2 increased by 1.98,0.92,1.27,0.52,0.37 and 0.17,5.31,1.90,2.97,1.26 times respectively(P<0.01).Com-pared with the group of 500μmol/L CS_2,TDNA%,TL,CL TM,OTM in the group of 1000 μmol/L CS_2,in creased by 0.55,0.49,0.16,1.18,0.76 times respectively(P<0.01).The result of regression analysis sbowed that regression coefficients between HIDNA%,TDNA%,TL TM,OTM and the doses were-13.78,13.78.0.05.4.38 and 3.23 respectively.Conclusions CS_2 exposure could induce DNA damage in the endometrial cells of mice at the phase of implantation.The degree of DNA damage increases with the increasing CS_2 concentration.CS_2 might affect the implantation of embryo by doing harm to the endometrial cells.