目的研究化学毒性物质氯化镉(cadmium chloride,CdCl2)对骨髓间充质干细胞(bone marrow mesenchymal stemcells,BMSCs)形态学、MTT检测结果、细胞凋亡及DNA稳定性的影响。方法用MTT法筛选CdCl2对BMSCs抑制作用的最佳浓度;将BMSCs(F3)分为对照组及CdCl2最佳浓度(200μmol/L)干预组,培养24 h后用MTT法检测BMSCs的生存情况,用流式细胞仪(flow-cytometer,FCM)检测细胞凋亡情况,用单细胞凝胶电泳技术(SCGE)检测BMSCs的DNA损伤情况。结果 CdCl2可对BMSCs在6.25~200μmol/L有明显的抑制作用,细胞给药12及24 h后,存在剂量-效应关系和时间-效应关系,CdCl2对BMSCs抑制作用最强的浓度为200μmol/L(24 h);显微镜下观察,对照组细胞呈成纤维细胞样贴壁生长,胞体透亮,折光性强;CdCl2诱导组细胞呈凸起回缩,细胞变小、变圆,贴壁不牢,悬浮细胞增多;与对照组比较,CdCl2诱导组细胞存活率明显降低,细胞凋亡明显增加,细胞的DNA拖尾率、平均尾长显著增加(P〈0.01)。结论 CdCl2在质量浓度为6.25~200μmol/L时对BMSCs有明显的抑制作用;对BMSCs抑制作用最强的浓度为200μmol/L(24 h),且可引起大鼠BMSCs凋亡及DNA的损伤。
In this study,we aim to investigate the effects of cadmium chloride(CdCl 2) on the morphology,MTT results,apoptosis,and DNA stability of bone marrow mesenchymal stem cells(BMSCs),and hope to provide a toxicological reference for the clinical application of BMSCs.The optimal concentration of CdCl 2 was selected using MTT assay.BMSCs(F3) were divided into two groups: control group and 200 μmol/L CdCl 2 induced group.After cultured for 24 h,the BMSCs of the two groups were tested by MTT assay for survival effect,by FCM for apoptotic ratios and by SCGE for the DNA damage.MTT assay results showed that CdCl 2 in 6.25-200 μmol/L had obvious inhibitive effects on BMSCs in dose-response relationship and time-effect relationship.The strongest inhibitive effect of CdCl 2 on rat BMSCs was appeared when the CdCl 2 concentration is 200 mmol/L and the stimulation duration is 24 h.In the CdCl 2 induced group,the cell viability significantly reduced,the apoptotic ratio was significantly increased,and the DNA damage(tail length,tail DNA %) are significantly increased,as compared with controls(P 0.01).All these results suggested that CdCl 2(work concentration is 6.25-200 μmol/L,optimal concentration is 200 μmol/L) has obvious inhibitive effects on BMSCs,and can induce apoptosis and DNA damage.