镉对体外培养鸡骨髓基质细胞成骨分化的毒性作用

Toxic effects of cadmium on osteogenic differentiation of chicken bone marrow stromal cells in vitro

骨是镉毒性作用的主要靶器官之一,但其对鸡骨髓基质细胞(bone marrow stromal cells,BMSCs)增殖和成骨分化的毒性作用仍不清楚。本研究利用差速贴壁纯化法获得鸡BMSCs,加入不同浓度镉处理不同时间,采用CCK-8法检测细胞增殖,碱性磷酸酶(alkaline phosphatase,ALP)和茜素红染色鉴定成骨分化,RT-PCR检测成骨相关基因(COL1、OSX、RUNX2、ALP、OCN、OPG、OPN、RANKL)表达变化。结果显示,1~10μmol/L的镉显著促进BMSCs增殖,20μmol/L的镉显著抑制其增殖(P<0.05);5μmol/L以上的镉可显著抑制ALP活性,并以浓度依赖方式极显著抑制成骨相关基因(COL1、OSX、RUNX2、ALP、OCN、OPG、OPN)mRNA表达,上调RANKL mRNA表达(P<0.01)。表明,一定浓度镉可抑制鸡BMSCs体外增殖及其向成骨细胞(OB)的分化。

Bone is one of the main target organs for cadmium toxicity,but this toxic effect on the proliferation and osteogenic differentiation of bone marrow stromal cells(BMSCs)is still unclear.In this study,chicken BMSCs were obtained by differential adhesion and were treated with different concentrations of cadmium for different lengths of time.Cell proliferation was detected by CCK-8,and osteogenic differentiation was identified by alkaline phosphatase(ALP)and alizarin red staining.Expression changes of osteoblast-related genes(COL1,OSX,RUNX2,ALP,OCN,OPG,OPN,RANKL)were detected by RT-PCR.The results showed that cadmium concentrations from 1~10μmol/L significantly promoted the proliferation of BMSCs,but those at and above 20μmol/L significantly inhibited this proliferation(P<0.05).Cadmium concentrations above 5μmol/L significantly inhibited ALP activity and the mRNA expression of osteogenic genes(COL1,OSX,RUNX2,ALP,OCN,OPG,OPN),and up-regulated the mRNA expression of RANKL dosage-dependently(P<0.01).In conclusion,cadmium inhibited chicken BMSCs proliferation and osteogenic differentiation in vitro.