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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. 培养C3H10T1/2细胞系，分为对照组、成脂诱导组和成脂诱导处理组（Y-27632+成脂诱导），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，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、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. To investigate the effect of Rho kinases inhibitor Y-27632 on proliferation and adipogenic differentiation of C3H10T1/2 mesenchymal stem cell. C3H10T1/2 cell were cultured in DMEM medium with 10% fetal bovine serum at 37℃ in 5% CO2 incubator. The experimental divided into control group, adipogenic group, adipogenic with Y-27632 group. The effects of Y-27632 on proliferation were detected by MTT. The adipogenic differentiation was characterized by oil red O staining. The differentiation of C3H10T1/2 cell to adipocyte was verified by expression of genes PPARγ and C/EBPα through RT-PCR. The results showed that Y-27632 obviously inhibited the proliferation of C3H10T1/2 cell in a dose-dependengt manner(P<0.05). High concentrations of Y-27632 significantly inhibited adipogenic differentiation of C3H10T1/2 cell (P<0.05). RT-PCR showed that the expression of PPARγ and C/EBPα in the 3rd, 5th day and 7th day were significantly inhibited by Y-27632 compared to the control (P<0.05). In summary, Y-27632 could inhibit the proliferation and adipogenic differentiation of C3H10T1/2 cell. |
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