脂联素对不同培养条件下成骨细胞与间充质干细胞中SDF-1/CXCR4表达影响的实验研究

目的研究不同培养条件下,脂联素对成骨细胞的基质细胞衍生因子(SDF-1)以及间充质干细胞CXCR4表达的影响。方法提取重组的脂联素蛋白,将10μg/m L脂联素作用于单独培养的MC3T3-E1以及C3H10T1/2细胞,Real-time定量PCR方法检测SDF-1/CXCR4表达变化;设计MC3T3-E1及C3H10T1/2细胞共培养体系,比较共培养对MC3T3-E1及C3H10T1/2细胞的SDF-1/CXCR4表达变化;将脂联素加入共培养体系,比较脂联素对共培养体系中MC3T3-E1及C3H10T1/2细胞SDF-1/CXCR4 mRNA表达变化。结果 Real-time定量PCR结果显示:单独培养条件下,脂联素促进了MC3T3-E1细胞SDF-1mRNA的表达;而在共培养体系中,脂联素则下调了MC3T3-E1细胞SDF-1mRNA的表达。无论单独培养还是共同培养,脂联素对C3H10T1/2细胞的CXCR4表达均起下调作用。结论脂联素对成骨细胞SDF-1的表达呈双向调节作用,而对间充质干细胞CXCR4则起下调作用。脂联素通过调节SDF-1/CXCR4影响间充质干细胞和成骨细胞的微环境。

An experimental study on the effect of adiponectin on SDF-1/CXCR4 expression of osteoblasts and mesenchymal stem cells in different culture systems

Objective To determine the effect of adiponectin on SDF-1/CXCR4 expression of MC3T3-E1 and C3H10T1/2 cells within different culture systems. Methods Cultured MC3T3-E1 and C3H10T1/2 cells were treated with and without adiponeetin re- spectively. Real-time quantitative PCR （qPCR） was used to detect SDF-1 and CXCR4 mRNA expressions. Then, a co-cuhure system with MC3T3-E1 cells in upper chamber and C3HIOT1/2 ceils in lower chamber was designed. After adiponectin treatment,cells were collected and SDF-1/CXCR4 mRNA expressions were examined. Results qPCR results showed that adiponectin increased SDF-1 mR- NA expression when MC3T3-E1 cells were cultured alone,while it decreased SDF-1 mRNA expression when MC3T3-E1 cells were cul- tured in co-cuhure system. Adiponectin inhibited CXCR4 mRNA expression in C3H10T1/2 cells,regardless of culture conditions. Con- clusions We concluded that adiponectin has dual effects on SDF-1 expression of MC3T3-E1 eells. Adiponectin influences mesenchy- mal stem cells and osteoblasts niche by regulating SDF-1/CXCR4 axis.