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目的探讨黄芪甲苷(Asd)对镉(Cd)致TM3细胞损伤的保护。方法 MTT细胞活性实验筛选Asd拮抗Cd的最佳保护浓度;流式细胞作TM3细胞凋亡分析;免疫组织化学检测连接蛋白43(connexin43,Cx43)的表达;电镜观察TM3细胞超微结构。结果 24 h、48 h MTT实验:对照组与Asd组细胞成活率差异无统计学意义,Cd组细胞成活率明显减弱且随Cd浓度增加减弱越明显,Cd+Asd组细胞成活率明显高于相应Cd组,Asd的最佳保护浓度为20 mg/L。流式细胞分析:对照组与Asd组24 h细胞凋亡率差异无统计学意义,Cd组随Cd浓度增加凋亡率逐渐增加,Cd+Asd组凋亡率均低于相应Cd组(P<0.01)。免疫组织化学:Cd处理后TM3细胞Cx43阳性产物表达的平均光密度值明显降低(P<0.01)、平均灰度值明显升高(P<0.01),Cd+Asd组阳性产物表达虽较对照组减弱但明显高于相应Cd组(P<0.01)。电镜观察:Cd处理后TM3细胞超微结构破坏明显,Cd+Asd组超微结构破坏较相应Cd组为轻。结论 Cd引起TM3细胞损伤并降低Cx43的表达,Asd有较好的保护效果。
Objective To investigate the protective effects of Astragaloside Ⅳ on the damage of TM3 cells induced by cadmium (Cd). Methods MTT activity was used to screen the best protection concentration of Asd against Cd. Flow cytometry was used to analyze TM3 cell apoptosis. Immunohistochemistry was used to detect the expression of connexin43 (Cx43). The ultrastructure of TM3 cells was observed by electron microscopy. Results MTT assay at 24 h and 48 h showed no significant difference in survival rate between control group and Asd group. The survival rate of cells in Cd group was significantly weakened and the more obvious with the increase of Cd concentration, the survival rate of Cd + Asd group was significantly higher than the corresponding Cd, Asd the best protection concentration of 20 mg / L. Flow cytometry analysis showed that there was no significant difference in 24 h apoptosis rate between control group and Asd group. The apoptosis rate of Cd + Asd group was higher than that of Cd group (P < 0.01). Immunohistochemistry: The average optical density of Cx43 positive products in TM3 cells was significantly decreased (P <0.01), the average gray value was significantly increased (P <0.01), while the positive expression of Cdx Asd in TM3 cells was higher than that in control group Attenuated but significantly higher than the corresponding Cd group (P <0.01). Electron microscopy showed that the ultrastructural destruction of TM3 cells was obvious after Cd treatment, and the ultrastructure destruction of Cd + Asd group was lighter than that of Cd group. Conclusion Cd can cause TM3 cell injury and reduce the expression of Cx43, Asd has a good protective effect.