POSTN通过ERK1/2信号通路诱导肺动脉平滑肌细胞增殖

Periostin Induces the Proliferation of Pulmonary Artery Smooth Muscle Cell Through ERK1/2 Signaling Pathway

目的探索骨膜蛋白(periostin,POSTN)诱导肺动脉平滑肌细胞(pulmonary artery smooth muscle cell,PASMC)增殖在缺氧诱导肺动脉高压(hypoxia-induced pulmonary hypertension,HPH)发病中的作用及其机制。方法取自雄性大鼠的原代PASMC暴露于缺氧环境模拟HPH,将细胞分为常氧组、缺氧组、缺氧+对照腺病毒转染组(Ad-shNC组)、缺氧+POSTN沉默腺病毒转染组(Ad-shPOSTN组)、POSTN组、POSTN+U0126组。采用α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)评估PASMC的纯度,CCK-8法检测细胞活力,Western blot法检测PASMC中POSTN、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、骨形态发生蛋白Ⅱ型受体(bone morphogenetic protein typeⅡreceptor,BMPR2)、磷酸化的细胞外信号调节激酶1/2(phosphorylated extracellular signal-regulated kinase 1/2,p-ERK1/2)及细胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2,ERK1/2)的蛋白表达水平。结果与常氧组比较,缺氧促进PASMC增殖和POSTN蛋白表达,抑制BMPR2蛋白表达。下调POSTN表达可抑制缺氧诱导的PASMC增殖,恢复BMPR2蛋白表达水平。此外,POSTN明显增加p-ERK1/2或ERK1/2的蛋白表达水平,增加PASMC增殖,而这些效应可被U0126阻断。结论在HPH病理机制中,POSTN促进PASMC增殖,其潜在的机制可能与调节BMPR2表达和活化ERK1/2信号通路有关。

Objective To explore the role and mechanism of periostin(POSTN)in the proliferation of pulmonary artery smooth muscle cell(PASMC)in hypoxia-induced pulmonary hypertension(HPH).Methods The primary PASMC from male rats were exposed to hypoxic environment to simulate HPH.The cells were divided into normoxia group,hypoxia group,hypoxia+control adenovirus transfected group(hypoxia+Ad-shNC group),hypoxia+POSTN silenced adenovirus transfected group(hypoxia+Ad-shPOSTN group),POSTN group,POSTN+U0126 group.α-smooth muscle actin(α-SMA)was used to evaluate the purity of PASMC,CCK-8 assay was performed to detect cell viability,and Western blot was utilized to detect the protein expression level of POSTN,proliferating cell nuclear antigen(PCNA),bone morphogenetic protein typeⅡreceptor(BMPR2),phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2),and extracellular signal-regulated kinase 1/2(ERK1/2)in PASMC.Results Compared with the normoxia group,hypoxia promoted the proliferation of PASMC and the expression level of POSTN protein,while inhibited the expression level of BMPR2 protein.Down-regulation of POSTN expression could inhibit hypoxia-induced proliferation of PASMC and restore BMPR2 protein expression level.In addition,POSTN significantly increased the protein expression ratio of p-ERK1/2 or ERK1/2,and the proliferation of PASMC,which could be blocked by U0126.Conclusion In the pathological mechanism of HPH,POSTN promotes the proliferation of PASMC,and its potential mechanism may be related to the regulation of BMPR2 expression and activation of ERK1/2 signaling pathway.