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观察低氧对大鼠肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)Periostin表达的影响及其相关信号转导机制。胶原酶I法原代培养PASMCs,经低氧(5%O2)分别处理PASMCs 2,6,12,24 h后,RT-PCR和Western blot法检测Periostin mRNA和蛋白表达。加入PI3K/Akt通路特异性抑制剂LY294002(10μmol/L)进行干预,Western blot分析比较不同条件下低氧处理24 h后大鼠PASMCs中Periostin和Akt/P-Akt的蛋白表达。结果表明,与常氧组比较,低氧处理6 h组、12 h组和24 h组Periostin mRNA和蛋白的表达均显著上升(P<0.05,P<0.01),低氧处理后的PASMCs中Periostin mRNA和蛋白的表达逐渐升高;低氧处理2 h组无显著差异(P>0.05)。用LY294002对PASMCs处理,并低氧24 h后,Periostin的表达被显著抑制(P<0.01),细胞P-Akt的表达下调(P<0.05),总Akt的蛋白表达没有明显差异(P>0.05)。推测低氧可诱导大鼠PASMCs中Periostin mRNA和蛋白的表达上调。低氧可能通过激活PI3K/Akt通路促进Akt的磷酸化,进而使Periostin在PASMCs中过表达,提示Periostin在低氧性PASMCs增殖过程中可能起着重要作用。
To observe the effect of hypoxia on Periostin expression in rat pulmonary artery smooth muscle cells (PASMCs) and its related signal transduction mechanism. PASMCs were cultured by collagenase I method. Periostin mRNA and protein expression were detected by RT-PCR and Western blot after treated with hypoxia (5% O2) for 2, 6, 12 and 24 h respectively. The PI3K / Akt pathway-specific inhibitor LY294002 (10μmol / L) was added for intervention. The protein expression of Periostin and Akt / P-Akt in PASMCs were detected by Western blot analysis under different conditions for 24 h. The results showed that Periostin mRNA and protein expression in 6 h, 12 h and 24 h hypoxia groups were significantly increased (P <0.05, P <0.01) compared with normoxia group. Periostin mRNA and protein expression gradually increased; hypoxia 2 h group no significant difference (P> 0.05). PASMCs were treated with LY294002 for 24 h and Periostin expression was significantly inhibited (P <0.01), P-Akt expression was down-regulated (P <0.05), and total Akt protein expression was not significantly different (P> 0.05 ). Hypoxia-inducible Periostin mRNA and protein expression in PASMCs were up-regulated. Hypoxia may promote the phosphorylation of Akt by activating the PI3K / Akt pathway, and then overexpress Periostin in PASMCs, suggesting Periostin may play an important role in the proliferation of hypoxic PASMCs.