BMP4通过PKA-钙蛋白酶-TGF-β1通路抑制肺动脉平滑肌细胞增殖和胶原合成  

BMP4 inhibits pulmonary artery smooth muscle cells proliferation and collagen synthesis via the PKA-Calpain-TGF-β1 pathway

作　　者：徐彪 吴巧玲[1] 倪吉祥[1] 罗干 赵天明[1] Xu Biao;Wu Qiaoling;Ni Jixiang;Luo Gan;Zhao Tianming(Department of Pulmonary and Critical Care Medicine,the People′s Hospital of China Three Gorges University,the First Hospital of Yichang,Yichang 443000,China)

机构地区：[1]三峡大学人民医院宜昌市第一人民医院呼吸与危重症医学科,宜昌443000

出　　处：《国际呼吸杂志》2022年第5期388-394,共7页International Journal of Respiration

基　　金：湖北省自然科学基金(2019CFB745)。

摘　　要：目的探究骨形成蛋白4(BMP4)在血小板源性生长因子(PDGF)诱导的肺动脉重塑中的作用及其机制。方法本研究为实验研究。使用细胞活力检测、蛋白活力测定、蛋白质印迹法、siRNA干扰技术等方法研究BMP4对肺动脉平滑肌细胞(PASMC)的作用。烟雾连续暴露12周小鼠构建肺动脉重塑的动物模型,免疫荧光检测肺动脉BMP4的表达。结果相较于PDGF组,PDGF+BMP4组能够抑制Ⅰ型胶原蛋白的合成(P<0.001)和钙蛋白酶活性(P<0.001)。相较于对照组,蛋白激酶A(PKA)的活性增加(P<0.001)。已知PKA是钙蛋白酶活性的负调控因子。使用PKA激活剂后,PDGF诱导的钙蛋白酶活性下降(P<0.01)。siRNA沉默PKA,由BMP4下调的钙蛋白酶活性得以恢复(P<0.001),并增加了Ⅰ型胶原蛋白的表达量(P<0.001)。与PDGF组相比,PDGF+BMP4抑制转化生长因子β1(TGF-β1)的活化(P<0.001)和Smad2/3的磷酸化(P<0.001)。在烟雾暴露的小鼠肺动脉中,BMP4蛋白表达降低(P<0.001)。结论BMP4通过激活PKA抑制钙蛋白酶-TGF-β1信号轴,导致PASMCⅠ型胶原蛋白合成下降。BMP4作为一种保护因素阻碍肺动脉重塑。Objective To explores the effects and mechanism of bone morphogenetic protein 4(BMP4)in platelet-derived growth factor(PDGF)-induced pulmonary artery remodeling.Methods This is an experimental study.The effects of BMP4 on pulmonary artery smooth muscle cells(PASMC)were investigated by cell viability assay,protein viability assay,western blotting,and siRNA interference technology.The mice were exposed to smoke for 12 weeks to set up an animal model of pulmonary artery remodeling,and the expression of BMP4 in the pulmonary artery was detected by immunofluorescence.Results The synthesis of collagen typeⅠ(P<0.001)and calpain activity(P<0.001)could be inhibited in the PDGF+BMP4 group compared with PDGF group.The activity of protein kinase A(PKA)increased(P<0.001)compared with the control group.PKA had been known to be a negative regulator of calpain activity.PDGF-induced calpain activity decreased after PKA activator(P<0.01).siRNA silencing of PKA restored calpain activity down-regulated by BMP4(P<0.001),and increased the expression collagen typeⅠ(P<0.001);PDGF+BMP4 inhibited the activation of transforming growth factor-β1(TGF-β1)(P<0.001)and the phosphorylation of Smad2/3(P<0.001)compared with the PDGF group.BMP4 protein expression decreased in the pulmonary arteries of smoke-exposed mice(P<0.001).Conclusions BMP4 inhibits the calpain-TGF-β1 signaling axis by activating PKA,causing the decrease of typeⅠcollagen synthesis in PASMC.BMP4 acts as a protective factor blocking pulmonary artery remodeling.

关 键 词：肺动脉 平滑肌细胞 血小板源性生长因子 骨形成蛋白4 钙蛋白酶 

分 类 号：R544.1[医药卫生—心血管疾病]