17β-雌二醇对骨髓间充质细胞向成骨细胞分化的诱导作用

Experimental study of 17β-estradiol induced bone marrow mesenchymal stem cells to osteoblast differentiation in vitr

目的 观察17β-雌二醇在骨髓间充质干细胞(BMSCs)向成骨细胞分化中的作用。方法 将诱导分化后的BMSCs分为对照组及17β-雌二醇组(1×10^-10、1×10^-9、1×10^-8 mol/L的17β-雌二醇),在诱导第7天,测定碱性磷酸酶(ALP)活性,细胞活力,Wnt1、Wnt10b及β-连环蛋白(β-catenin)表达量;在诱导第14天,测定各组细胞Ⅰ型胶原酶(COIⅠ)mRNA含量;在诱导第21天,测定各组细胞成骨矿化能力,骨桥蛋白(OPN)及骨钙素(OCN)mRNA含量。结果 与对照组比较,1×10^-10、1×10^-9、1×10^-8 mol/L的17β-雌二醇能显著提高细胞活力[(0.41±0.03)比(0.45±0.04)、(0.51±0.05)、(0.57±0.05)](P<0.01),ALP活性[(0.23±0.02)比(0.34±0.02)、(0.42±0.03)、(0.48±0.05)](P<0.01),增加钙结节数量(P<0.01),上调COIⅠ、OPN、OCN mRNA表达量及Wnt1、Wnt10b及β-catenin表达量(P<0.01)。结论 17β-雌二醇可能通过激活Wnt/β-catenin信号通路促进BMSCs向成骨细胞分化。

Objective To explore experimental study of 17β-estradiol induced bone marrow mesenchymal stem cells (BMSCs) to osteoblast differentiation in vitro.Methods BMSCs was divided into control group, 17β-estradiol group (1×10-10, 1×10-9, 1×10-8 mol/L 17β-estradiol). The activity of alkaline phosphatase (ALP), cell viability, the expression of Wnt 1, Wnt 10b and β-catenin were measured on the 7th day after induction. The expression of collagenase type Ⅰ (COIⅠ) mRNA was measured on the 14th day after induction. The osteogenic mineralization ability, the expression of osteopontin (OPN) and osteocalcin (OCN) mRNA content were measured on the 21st day after induction.Results Compared with the control group, the activity of ALP [ (0.23±0.02) vs. (0.34±0.02), (0.42±0.03), (0.48±0.05)] (P<0.01), cell viability [ (0.41±0.03) vs. (0.45±0.04), (0.51±0.05), (0.57±0.05)] (P<0.01), the number of calcium nodule (P<0.01), the expression of COI Ⅰ, OPN and OCN mRNA, the expression of Wnt1, Wnt10band β-cateninwas increased in 10-10, 10-9, 10-8 mol/L 17β-estradiol group (P<0.01).Conclusion 17β-estradiol might promote BMSCs differentiation into osteoblast via activating Wnt/β-catenin signal pathway.