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目的探讨Ⅰ型去乙酰化酶抑制剂(histone deacetylase inhibitor,HDACi)MS-275对间充质干细胞(mesen-chymal stem cells,MSCs)成脂分化的影响。方法利用噻唑蓝(MTT)和碘化乙啶(PI)染色结合流式细胞术(FCM)分别检测不同浓度MS-275对C3H/10T1/2细胞活性和细胞周期的影响;油红O染色分析MS-275对C3H/10T1/2细胞成脂分化能力的影响;同时应用实时定量PCR(real-time PCR)检测MS-275对成脂分化标志物:脂结合蛋白(aP2)、围脂素(perilipin)、脂联素(Adipoq),以及成脂分化关键转录因子:过氧化物酶体增殖物激活受体-γ2(PPAR-γ2)mRNA转录水平的影响。结果随着MS-275浓度升高,其对C3H/10T1/2细胞的抑制率也随着增高,IC50约为8μmol/L(P<0.05);流式结果表明,MS-275将C3H/10T1/2细胞周期阻滞在G0/G1期;0.5μmol/L的MS-275明显减少C3H/10T1/2细胞的脂滴积累;同时成脂分化标志物aP2、perilipin、Adipoq及成脂关键转录因子PPAR-γ2的转录水平也显著降低(P<0.05)。结论 MS-275减弱了间充质干细胞成脂分化的能力,表明抑制Ⅰ型HDACs的活性可部分抑制MSCs向成脂分化。
Objective To investigate the effect of type I deacetylase inhibitor (HDACi) MS-275 on adipogenic differentiation of mesenchymal stem cells (MSCs). Methods MTT, PI staining and flow cytometry (FCM) were used to detect the effects of different concentrations of MS-275 on C3H / 10T1 / 2 cell viability and cell cycle. Oil red O staining MS-275 on adipogenic differentiation ability of C3H / 10T1 / 2 cells was detected by real-time PCR. The effects of MS-275 on adipogenic differentiation markers such as lipid binding protein (aP2) perilipin, Adipoq and the key transcription factor of adipogenic differentiation: the transcription level of peroxisome proliferator activated receptor-γ2 (PPAR-γ2) mRNA. Results The inhibitory rate of C3H / 10T1 / 2 cells increased with the increase of MS-275 concentration and the IC50 was about 8μmol / L (P <0.05). The results of flow cytometry showed that MS- / 2 cell cycle arrest in G0 / G1 phase; 0.5μmol / L MS-275 significantly reduced the lipid droplets accumulation in C3H / 10T1 / 2 cells; at the same time adipogenic differentiation markers aP2, perilipin, Adipoq and adipogenic key transcription factor The transcription level of PPAR-γ2 was also significantly decreased (P <0.05). Conclusion MS-275 attenuates the ability of mesenchymal stem cells to differentiate into adipogenic cells, indicating that inhibiting the activity of type I HDACs can partially inhibit the differentiation of MSCs into adipogenic cells.