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目的观察Cr3+对成骨细胞SaO2的细胞毒性以及在Cr3+的作用下对成骨细胞Tnfrsf17基因的表达的影响。方法体外培养SaO2成骨细胞,MTT法检测细胞活力,ALP试剂盒检测成骨细胞碱性磷酸酶的分泌情况,Von Kossa试剂盒检测细胞钙结节形成。通过Rt-PCR及Western检测Cr3+对Tnfrsf17表达的影响。结果在Cr3+的刺激作用下,与对照组相比,成骨细胞在相应时间节点细胞活性和钙结节形成明显受限,ALP的分泌呈时间与剂量依赖性的降低。在Cr3+的刺激作用下,0.1、1.3和150 mg各干预组与对照组相比,Tnfrsf17的基因表达水平及蛋白表达水平在24、48和72 h后均有所下降。结论 Cr3+对成骨细胞有细胞毒性而且可以下调Tnfrsf17基因及其蛋白产物的表达。
Objective To investigate the cytotoxicity of Cr3 + on osteoblasts and the effect of Cr3 + on the expression of Tnfrsf17 in osteoblasts. Methods SaO2 osteoblasts were cultured in vitro. Cell viability was assayed by MTT assay. ALP kit was used to detect osteoblasts alkaline phosphatase secretion. Von Kossa kit was used to detect the formation of calcium nodules. The effect of Cr3 + on the expression of Tnfrsf17 by Rt-PCR and Western blotting. Results Compared with the control group, the cell viability and calcium formation at the corresponding time points were significantly inhibited by Cr3 + stimulation, and the secretion of ALP was decreased in a time-and dose-dependent manner. Under the stimulation of Cr3 +, the gene expression and protein expression of Tnfrsf17 decreased at 24, 48 and 72 h after treatment with 0.1, 1.3 and 150 mg of each compared with the control group. Conclusion Cr3 + is cytotoxic to osteoblasts and down-regulates Tnfrsf17 gene and its protein product expression.