阿托伐他汀对体外培养的大鼠骨髓间充质干细胞CXC趋化因子受体4表达及迁移能力的影响

Effect of atorvastatin on the expression of chemokine receptor-4 and migration ability of rat bone marrow mesenchymal stem cells cultivated in vitro

目的观察不同浓度阿托伐他汀（ATV）对体外培养的大鼠骨髓间充质干细胞（BMSC）中 CXC趋化因子受体4（CXCR4）表达及迁移能力的影响。方法分离培养鉴定大鼠BMSC。将第4-6代细胞分为对照组和ATV处理0.1 nmol/L组、1.0 nmol/L组、10.0 nmol/L组、100.0 nmol/L组、1 000.0 nmol/L组。ATV处理12 h后，细胞免疫荧光法、蛋白免疫印迹法（Western blot）检测各组细胞中CXCR4蛋白表达；实时定量聚合酶链反应（RT-PCR）检测各组细胞中CXCR4 mRNA表达；Transwell小室法检测细胞迁移能力。组间细胞中mRNA、蛋白表达和细胞迁移能力比较行独立样本t检验。结果细胞免疫荧光法检测结果显示，1.0 nmol/L组、10.0 nmol/L组细胞中CXCR4蛋白表达量较对照组、0.1 nmol/L组、100.0 nmol/L组、1 000.0 nmol/L组明显增高；RT-PCR、Western blot检测结果显示，10.0 nmol/L组细胞中CXCR4 mARNA、蛋白表达均较1.0 nmol/L组和对照组明显增高，差异有统计学意义（FmARNA=20.36，P=0.005；F蛋白=33.17，P=0.009）；Transwell小室法检测结果显示，10.0 nmol/L组细胞迁移能力较1.0 nmol/L和对照组明显提高，差异有统计学意义（F=43.77，P=0.000）。结论较低浓度ATV可呈剂量依赖性促进体外培养的BMSC中CXCR4表达并提高其迁移能力。

ObjectiveTo investigate the effects of migration and expression from chemokine receptor 4 （chemokine receptor-4, CXCR4） of rat bone marrow mesenchymal stem cells （BMSCs） which were pretreated by atorvastatin （ATV） in vitro.MethodsIsolated, cultivated, identified the BMSCs, pretreated P4-P6 of BMSCs with different concentrations of ATV for 12 hours. The experimental group was divided into control group, 0.1 nM/L （group 0.1 nM）, 1 nM/L （1 nM group）, 10 nM/L （10 nM group）, 100 nM/L （100 nM group）, 1 000 nM/L （1 000 nM group）. The mRNA and protein of CXCR4 were determined by real time-polymerase chain reaction and Western blot. Immunofluoreseence assay were used to detect the expression levels of CXCR4. The migration ability of BMSCs were measured by transwell chamber.ResultsImmunofluoreseence assay showed the protein level of CXCR4 of group 1 nM and 10 nM were significantly higher than the other group. RT-PCR and Western blot showed the protein and mRNA level of CXCR4 in 10 nM was higher than that in group 1 nM. The migration ability of group 10 nM was higher than 1 nM and control group.ConclusionsATV can be dose-dependent promote expression levels of CXCR4 of BMSCs cultivated in vitro.