PPAR δ激动剂GW501516对大鼠骨髓基质干细胞分化的作用

The effect of PPAR δ agonist-GW501516 on the differentiation of rats bone marrow stromal stem cells

目的探讨过氧化物酶体增殖激活受体δ(PPARδ)激动剂GW501516对大鼠骨髓基质干细胞(BMSCs)向成脂和成骨方向分化的作用。方法全骨髓培养法培养大鼠原代BMSCs后采用差速贴壁法纯化,流式细胞仪鉴定其细胞表面标志物,实验分为PPARδ激动剂组和对照组,分别进行成骨和成脂方向诱导培养,荧光定量PCR检测成骨细胞分化标志物(碱性磷酸酶、骨钙素)和脂肪细胞分化标志物(脂肪酸结合蛋白2、脂连素)mRNA的表达,油红O染色检测脂肪细胞分化,茜素红染色检测成骨细胞矿化情况。结果与对照组比较,PPARδ激动剂促进BMSCs向成骨细胞方向分化标志物表达,抑制向脂肪细胞方向分化标志物表达。茜素红染色显示PPARδ激动剂组细胞矿化结节较对照组增加,油红染色显示PPARδ激动剂组脂肪小滴明显减少。结论PPARδ激动剂促进BMSCs向成骨方向分化,抑制其向成脂方向分化。

Objective To investigate the effect of peroxisome proliferators-activated receptor δ （ PPAR δ） agonist GW501516 on the adipogenic and osteogenic differentiation of rats bone marrow stromal stem cells （BMSCs）. Methods Rat BMSCs were cultured using whole marrow adhesion method. The biomarkers on the cell membrane were identified using flow cytometry. BMSCs were randomly divided into PPAR δ agonist group and control group. The expression of osteoblast differentiation markers （ alkaline phosphatase, osteocalcin） and adipocyte differentia- tion markers （ap2, fat vitronectin） were detected using Real-time PCR after osteogenic and adipogenic induced culture. Mineralized nodules and lipid droplets were detected using alizarin red and oil red O staining. Results Compared with the control group, cells in PPAR δ agonist group had more expression of osteoblast differentiation markers and less expression of adipocyte differentiation markers. There were more mineralized nodules and fewer lipid droplets in the PPAR δ agonist group. Conclusion PPAR δ agonist can promote osteogenic differentiation of BMSCs, and inhibit their differentiation into adipocytes direction.