葛根素抑制间充质干细胞成脂肪分化的作用及机制

Effects and Mechanism of Puerarin in Inhibiting Adipogenic Differentation of Mesenchymal Stem Cells

目的:探讨葛根素对骨髓间充质干细胞(bone-marrow derived mesenchymal stem cells,BMSCs)成脂肪分化的抑制作用与分子机制。方法:Percoll梯度密度离心法分离BMSCs;采用BMSCs特异性无血清培养基进行培养及传代;流式细胞术鉴定BMSCs表面特异性抗原表达;0.2μM和0.5μM葛根素分别联合成脂肪分化诱导培养基BMSCs向脂肪细胞分化;油红染液在第12天鉴定脂肪细胞形成情况;real-time PCR和western blot分别鉴定成脂肪分化相关因子PPAR-γ、CEBP-α及CEBP-β的表达;观察0.2μM和0.5μM葛根素对成脂肪分化相关Wnt信号通路的影响。结果:葛根素可显著抑制BMSCs向脂肪细胞分化,并呈浓度依赖性(P<0.05),成脂肪诱导分化12天时,未加葛根素的BMSCs中PPAR-γ、CEBP-α及CEBP-β表达水平高于加葛根素组;葛根素组BMSCs胞内β-cateinin表达受到明显抑制。结论:葛根素可以明显抑制BMSCs向成脂肪细胞转化。

Objective: To explore the inhibitory effects of puerarin on adipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs) and its molecular mechanism. Methods: Percoll gradient density centrifugation method was used to separate BMSCs;BMSCs specific serum-free medium for the cultivation and the subculture;flow cytometry to identify the expressions of BMSCs surface specific antigen;0.2 μM and 0.5 μM puerarin combined with adipogenic differentiation-induced medium respectively to induce adipogenic differentiation of BMSCs respectively;oil red dye solution to identify the formation of adipocytes at the 12 th day;real-time PCR and western blot were adopted to measure the expressions of PPAR-γ, CEBP-α and CEBP-β in m RNA and protein levels;to observe the influence of 0.2 μM and 0.5 μM puerarin on adipogenic differentiation related wnt signaling pathway. Results: Puerarin could inhibit adipogenic differentiation of BMSCs remarkably, in a dose-dependent manner(P<0.05),After inducing for 12 days, the expressions of PPAR-γ, CEBP-α and CEBP-β in BMSCs without adding puerarin were higher than these of puerarin group;the expressions of β-cateinin in BMSCs were inhibited obviously in puerarin group. Conclusion: Puerarin could notably inhibit adipogenic differentiation of BMSCs.