亚硒酸钠与纳米硒对镉致小鼠睾丸间质细胞毒性的干预作用

PROTECTIVE EFFECTS OF SODIUM SELENITE AND NANO-SELENIUM ON CADMIUM-INDUCED TESTICULAR INTERSTITIAL CELL TOXICITY IN MICE

目的 比较亚硒酸钠(Se)和纳米硒(Nano-Se)对镉(Cd)致小鼠睾丸间质细胞毒性的干预作用。方法将小鼠睾丸间质细胞分成四组,分别是对照组、镉染毒组(Cd20μmol/L)、硒+镉染毒组(Se+Cd)、纳米硒+镉染毒组(Nano-Se+Cd),每组至少3个样本,重复3次以上。CCK-8检测细胞活性,Hoechst染色检测细胞凋亡形态变化,流式细胞仪分析细胞凋亡率,Westernblot测定蛋白的相对表达含量。结果 与Cd染毒组比较,Se组和Nano-Se组细胞活性明显上升(P<0.01);当Se和Nano-Se的浓度达20μmol/L时,Se组和Nano-Se组细胞活性最高(P<0.01),且Nano-Se与相同浓度的Se比较,Nano-Se组的细胞活性较高(P<0.01)。细胞形态观察发现,Cd染毒组细胞皱缩变圆,体积变小,细胞间距变大,荧光染色细胞核染色较多;与Cd染毒组比较,Se组和Nano-Se组生长形态较好,荧光染色细胞核染色较少,Nano-Se组比Se组生长情况更佳。Cd染毒组细胞凋亡率显著高于对照组(P<0.01),而与Cd染毒组比较,Se组和Nano-Se组细胞凋亡率明显降低(P<0.01);且与Se组比较,Nano-Se组细胞凋亡率更低(P<0.01)。与对照组比较,Cd染毒组TM3细胞Bax/Bcl-2比值明显增大(P<0.01);与Se组比较,Nano-Se组的Bax/Bcl-2比值更低(P<0.05)。结论 Nano-Se与Se比较对Cd致TM3细胞毒性干预作用更好,可能与Nano-Se能更好抑制镉引起的TM3细胞凋亡有关。

Objective To compare the protective effects of sodium selenite(Se) and nano-selenium(Nano-Se) on cadmium(Cd)-induced testicular interstitial cell toxicity in mice.Methods Mouse testicular Leydig cells were divided into four groups:control group,cadmium exposure group(Cd 20 μmol/L),selenium+cadmium group(Se+Cd),and nanoselenium+cadmium group(Nano-Se+Cd).Cell viability was detected by CCK-8 Kit,and apoptotic morphological changes were detected by Hoechst staining.Flow cytometry was used to analyze cell apoptosis rate and the relative protein levels were determined by Western blot.Results In contrast to the Cd exposure group,the cell viability in Se and Nano-Se groups was significantly higher(P<0.01).When the concentration of the Se and Nano-Se reached 20 μmol/L,the cell viability of the Se group and Nano-Se group was the highest(P<0.01).At the same concentration the cell viability of the Nano-Se group was higher than that of the Se group(P<0.01).The cell morphology observation showed that the cells in the Cd exposure group were wrinkled and rounded,smaller in size,with larger cell spacing and more fluorescent staining of nuclei.The cell in the Se and Nano-Se groups had better growth morphology and less fluorescent staining of nuclei by comparison with the Cd exposure group,and the Nano-Se group had better cellular growth than the Se group.The apoptosis rate in the Cd exposure group increased dramatically in comparison with the control group(P<0.01),while the apoptosis rate decreased considerably in the Se and Nano-Se groups(P<0.01).Meanwhile,the apoptosis rate of the Nano-Se group was lower than that of the Se group(P<0.01).Compared with the control group,the Bax/Bcl-2 ratio increased significantly in the Cd-exposured TM3cells(P<0.01).Compared with the Se group,the Bax/Bcl-2 ratio in the Nano-Se group was decreased(P<0.05).Conclusion Nano-Se was better than Se in terms of the protective effects on Cd-induced TM3 cell toxicity,which was probably related to the fact that Nano-Se could better inhibit cadmium-induced apoptosis in TM3 cells.