Rho激酶抑制剂Y-27632对C3H10T1/2细胞增殖与成脂分化的作用

探究Rho激酶抑制剂Y-27632对间充质干细胞（mesenchymal stem cells,MSCs）C3H10T1/2增殖和成脂分化的影响.实验分为对照组、成脂诱导组和Y-27632处理组（Y-27632+成脂诱导）. 利用MTT检测细胞增殖情况，油红O染色,异丙醇萃取法检测细胞成脂分化情况，半定量RT-PCR检测过氧化物酶体增殖物激活受体γ(peroxisome proliferator activiated receptor γ, PPARγ)和CCAAT增强子结合蛋白α (CCAAT enhancer binding protein α, C/EBPα)基因表达. 结果表明，Y-27632能够显著抑制C3H10T1/2细胞的增殖(P<0.05)，并呈一定的浓度依赖性；高浓度Y-27632对C3H10T1/2细胞成脂分化具有显著抑制作用（P<0.05）；半定量RT-PCR结果显示，成脂诱导处理组PPARγ和C/EBPα表达量在第3 d、5 d和7 d显著低于成脂诱导组（P<0.05）. 综上所述，Y-27632能够抑制C3H10T1/2细胞增殖与成脂分化.

Effect of Rho Kinases Inhibitor Y-27632 on Proliferation and Adipogenic Differentiation of Mesenchymal Stem Cell C3H10T1/2

To investigate the effect of Rho kinases inhibitor Y-27632 on the proliferation and adipogenic differentiation of mesenchymal stem cells, the C3H10T1/2 cells were cultured in DMEM medium with 10% fetal bovine serum at 37 ℃ in 5% CO2 incubator and divided into the control, adipogenic and Y-27632 groups. The cell proliferation was detected by MTT; the adipogenic differentiation was characterized by oil red O staining. The C3H10T1/2 cell differentiation into adipocytes was also examined by the gene expression of PPARγ and C/EBPα by RT-PCR. The results showed that Y-27632 inhibited the proliferation of C3H10T1/2 cell in a dose-dependent manner (P<0.05). High concentrations of Y-27632 inhibited adipogenic differentiation of C3H10T1/2 cell (P<0.05). The RT-PCR assays showed that the PPARγ and C/EBPα expression were inhibited by Y-27632 at day 3, 5 and 7 during differentiation as compared to the control (P<0.05). In summary, Y-27632 is able to inhibit the proliferation and adipogenic differentiation of C3H10T1/2 cell.