黄芪多糖对低氧环境中BMSCs成脂分化的影响

Effects of Astragalus polysaccharides on adipogenic differentiation of bone marrow mesenchymal stem cells in low oxygen environment

摘要:

探讨黄芪多糖 (APS) 对低氧环境中骨髓间充质干细胞 (BMSCs) 成脂诱导分化的影响。采用四甲基偶氮唑盐 (Methyl thiazolyl tetrazolium，MTT) 法筛选促进BMSCs增殖的最佳APS浓度，干预不同氧浓度下 (3%、6%、10%和20%) 成脂诱导培养的BMSCs，通过油红O染色观察细胞内脂滴形成及Real-time PCR和Western blotting，检测成脂相关基因过氧化物酶体增殖物激活受体γ2 (PPAR-γ2) 和脂蛋白脂肪酶 (LPL) 的mRNA和蛋白水平。结果表明，与对照组相比，40 μg/mL APS能显著促进不同氧浓度下BMSCs的增殖 (P<0.05)；含有40 μg/mL APS的成脂诱导剂能提升低氧环境中BMSCs内脂滴含量及PPAR-γ2和LPL的蛋白和mRNA水平，在氧浓度为10%时其促进作用较显著 (P<0.05)，差异有统计学意义。40 μg/mL APS具有促进低氧环境中BMSCs增殖和成脂诱导分化的作用，其促分化作用与细胞培养的氧环境相关。

关键词:骨髓间充质干细胞，低氧环境，黄芪多糖，成脂分化

Abstract:

To study the effect of Astragalus polysaccharide (APS) on adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) cultured in hypoxic environment. The optimal APS concentration, which could promote the proliferation of BMSCs, was screened by methyl thiazolyl tetrazolium method. The concentration was used to intervene in BMSCs-induced by adipogenic differentiation fluid growing in different oxygen concentrations (3%, 6%, 10% and 20%). The formation of lipid droplets in the BMSCs-intervened was observed by oil red O staining under the optical microscope. The mRNA and protein levels of the lipid relating genes peroxisome proliferator activated receptor gamma 2 (PPAR-γ2) and lipoprotein lipase (LPL) were detected by Real-time PCR and Western blotting, respectively. The results showed that, comparing with the control group, 40 μg/mL APS could significantly promote the proliferation of BMSCs under low oxygen concentration. A large amount of lipid droplets existed in BMSCs growing in the adipogenic inducing fluid containing 40 μg/mL APS and the hypoxic environment, and the protein and mRNA levels of PPAR-γ2 and LPL also raised. It was worth noting that the phenomenon was more significant in 10% oxygen concentration, and the difference was statistically significant (P<0.05). 40 μg/mL APS had effect on promoting the proliferation and adipogenic differentiation of BMSCs cultured in hypoxic environment, and the effect was related to the concentration of oxygen of BMSCs-cultured.

Key words:bone mesenchymal stem cells, low oxygen environment, Astragalus polysaccharides, adipogenic differentiation