他克莫司对小鼠骨髓间充质干细胞增殖及成骨分化的影响

Effects of immunosuppressant tacrolimus on proliferation and osteogenic differentiation of mouse BMSC

目的:研究他克莫司对小鼠骨髓间充质干细胞(BMSC)增殖及成骨分化的影响。方法:取小鼠BMSC分为3组,低、高剂量他克莫司组分别用含不同浓度(50、500 nmol/L)他克莫司的成骨诱导培养基培养,空白对照组用不含他克莫司的成骨诱导培养基培养。培养3 d,通过CCK-8法检测细胞的增殖能力;培养5、10 d,采用qRT-PCR检测成骨分化相关基因Runt相关转录因子2(Runx2)、锌指结构转录因子(Osterix)、碱性磷酸酶(Alp)、骨钙蛋白(Ocn)mRNA的表达水平;培养10、14 d,用ALP染色和茜素红染色检测细胞的成骨分化能力。结果:与空白对照组相比,低、高剂量他克莫司组BMSC增殖能力增强,成骨分化相关基因Runx2、Osterix、Alp、Ocn表达上调,ALP活性增强,矿化结节形成增多(P<0.05)。结论:他克莫司可促进小鼠BMSC的增殖和成骨分化。

Aim:To evaluate the effects of tacrolimus on the proliferation and osteogenic differentiation of mouse bone marrow mesenchymal stem cells(BMSC).Methods:Mouse BMSC were collected and cultured with different concentrations of tacrolimus(50 and 500 nmol/L).BMSC in control group were cultured with osteogenic differentiation media without tacrolimus.After 3 days cell culture,CCK-8 assay was used to detect cell proliferation.After 5 days and 10 days,the gene expression levels of osteogenic differentiation related markers runt-related transcription factor-2(Runx2),zinc finger structure transcription factor(Osterix),alkaline phosphatase(Alp)and osteocalcin(Ocn)were evaluated by qRT-PCR.ALP staining and alizarin red staining were respectively performed after 10 days and 14 days so as to detect the osteogenic differentiation ability.Results:Both 50 nmol/L and 500 nmol/L tacrolimus enhanced cell proliferation.BMSC in tacrolimus treated groups exhibited higher mRNA expressions of osteogenic differentiation related markers,enhanced ALP activity and mineralization nodes formation(P<0.05).Conclusion:Tacrolimus promotes the proliferation and osteogenic differentiation of mouse BMSC.