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目的探讨血小板源性生长因子(Platelet derived growth factor,PDGF)-BB诱导骨髓间充质干细胞(Bone marrow-derived mesenchymal stem cells,BMSCs)向血管平滑肌样细胞(Vascular smooth muscle-like cells,VSMLCs)分化过程中Periostin的表达及其在促VSMLCs分化中的作用。方法采用全骨髓贴壁培养法分离和培养大鼠BMSCs,取第2代BMSCs分为2组:诱导Ⅰ组(用50 ng/ml PDGF-BB单独向VSMLCs诱导)和诱导Ⅱ组(加入地塞米松1μmol/L、胰岛素1μmol/L、吲哚美辛1μmol/L、3-异丁基-1甲基黄嘌呤0.5 mmol/L,向脂肪样细胞诱导),以大鼠胸大动脉平滑肌细胞作为阳性对照。分别于诱导后7 d和14 d,采用RT-PCR检测细胞中平滑肌肌动蛋白(SMα-actin)、平滑肌肌球蛋白重链(SM MHC)、平滑肌肌钙结合蛋白(SM Calponin)和Periostin mRNA的转录水平,Western blot检测Periostin蛋白的表达水平。结果诱导Ⅰ组细胞的SMα-actin、SM MHC、SMCalponin和Periostin基因mRNA及Periostin蛋白的表达水平14 d比7 d显著增强,且差异有统计学意义(P<0.05);14 d与阳性细胞相比,差异无统计学意义(P>0.05);未诱导组及诱导Ⅱ组在14 d均无表达。结论 PDGF-BB(50 ng/ml)能够单独诱导BMSCs向VSMCs分化,Periostin在此过程中起重要作用。
Objective To investigate the differentiation of Bone marrow-derived mesenchymal stem cells (BMSCs) into vascular smooth muscle-like cells (VSMLCs) by platelet derived growth factor (PDGF) -BB. Expression of Periostin and its role in promoting differentiation of VSMCs in. Methods BMSCs were isolated and cultured by whole bone marrow adherent culture. The second passage BMSCs were divided into two groups: Induction group Ⅰ (induced by VSMCs with 50 ng / ml PDGF-BB alone) and induction group Ⅱ 1 μmol / L of insulin, 1 μmol / L of insulin, 1 μmol / L of indomethacin, and 0.5 mmol / L of 3-isobutyl-1-methylxanthine) were injected into adipose- Control. The expression of SMα-actin, SM MHC, SM Calponin and Periostin mRNA were detected by RT-PCR at 7 and 14 days after induction respectively. The transcription level of Periostin protein was detected by Western blot. Results The expression of SMα-actin, SM MHC, SMCalponin and Periostin mRNA and Periostin mRNA and protein in groupⅠ were significantly higher than those on day 7 after 14 d (P <0.05) The difference was not statistically significant (P> 0.05). The non-induced group and induced group Ⅱ were not expressed on the 14th day. Conclusion PDGF-BB (50 ng / ml) can induce the differentiation of BMSCs into VSMCs alone. Periostin plays an important role in this process.