BMP-7/Smads通路参与EndoMT在大鼠HHPH中的作用  

Role of BMP-7/Smads pathway in regulation of EndoMT in rats with HHPH

作　　者：张晶晶[1,2] 武垣伶 黄丹娜 高慧[1] 楼国强[1] 周卓琳 Lingfang SHI 王万铁[1] ZHANG Jing-jing;WU Yuan-ling;HUANG Dan-na;GAO Hui;LOU Guo-qiang;ZHOU Zhuo-lin;Lingfang SHI;WANG Wan-tie(Department of Pathophysiology,Wenzhou Medical University,Wenzhou 325035,China;Department of Pathology,Second Affiliated Hospital,Wenzhou Medical University,Wenzhou 325035,China;Division of Pulmonary and Critical Care Medicine,Stanford University Medical School,Stanford CA 94305-5236,USA.)

机构地区：[1]温州医科大学病理生理学教研室,浙江温州325035 [2]温州医科大学附属第二医院病理科,浙江温州325035 [3]Division of Pulmonary and Critical Care Medicine,Stanford University Medical School,Stanford,CA 94305-5236,USA

出　　处：《中国病理生理杂志》2020年第2期316-322,共7页Chinese Journal of Pathophysiology

基　　金：浙江省中医药重点研究计划(No.2018ZZ018);温州市高层次人才创新技术重点资助项目(No.2016-07);温州市引智项目(No.604090355/033)

摘　　要：目的:探讨骨形态发生蛋白(BMP)-7/Smads通路参与的内皮-间充质转化(EndoMT)在大鼠低氧高二氧化碳性肺动脉高压(HHPH)中的作用。方法:将大鼠肺动脉内皮细胞(RPAECs)分为常氧对照(Con)组、低氧高二氧化碳(HH)组、BMP受体激动剂rhBMP-7组、BMP受体抑制剂DMH-1组和溶剂DMSO组。各组给予相应的药物后,Con组置于常氧培养箱中培养,其余4组置于低氧高二氧化碳培养箱中培养。采用免疫荧光标记法观察细胞CD31和α-平滑肌肌动蛋白(α-SMA)的表达水平;RT-PCR和Western blot法分别检测各组细胞α-SMA、CD31和Smad1/5/8的mRNA和蛋白表达水平;CCK-8法检测细胞活力;Transwell小室法检测细胞的迁移水平。结果:与Con组相比,HH组α-SMA的mRNA和蛋白表达水平升高,而CD31的mRNA和蛋白表达水平、Smad1/5/8的mRNA和p-Smad1/5/8的蛋白表达水平降低,同时细胞活力降低而迁移增多(P<0.05)。与HH组相比,rhBMP-7组α-SMA的mRNA和蛋白表达水平降低,而CD31的mRNA和蛋白表达水平、Smad1/5/8的mRNA和p-Smad1/5/8的蛋白表达水平升高,同时细胞活力升高而迁移减少(P<0.05)。与rhBMP-7组相比,DMH-1组α-SMA的mRNA和蛋白表达水平升高,而CD31的mRNA和蛋白表达水平及Smad1/5/8的mRNA和p-Smad1/5/8的蛋白表达水平降低,同时细胞活力降低而迁移增多(P<0.05)。结论:低氧高二氧化碳可促进RPAECs发生EndoMT,从而促进HHPH的发展,其机制可能与抑制BMP-7/Smads通路有关。AIM:To investigate the role of bone morphogenetic protein 7(BMP-7)/Smads pathway in the re-gulation of endothelial-mesenchymal transition(EndoMT)in rats with hypoxia-hypercapnia pulmonary hypertension(HHPH).METHODS:The rat pulmonary artery endothelial cells(RPAECs)were divided into normoxic control(Con)group,hypoxia-hypercapnia(HH)group,BMP receptor agonist rhBMP-7 group,BMP receptor inhibitor DMH-1 group and solvent DMSO group.After given the corresponding drugs in each group,the cells in Con group were cultured in a normal-oxygen incubator,and the cells in the remaining 4 groups were cultured in a low-oxygen and high-carbon-dioxide incubator.The expression levels of CD31 andα-smooth muscle actin(α-SMA)were observed by immunofluorescence staining.The mRNA and protein expression levels ofα-SMA,CD31 and Smad1/5/8 were detected by RT-PCR and Western blot,respectively.The cell viability was measured by CCK-8 assay.The cell migration ability was detected by Transwell chamber assay.RESULTS:Compared with Con group,the expression ofα-SMA at mRNA and protein levels in HH group was increased,the expression levels of CD31 mRNA and protein,Smad1/5/8 mRNA and p-Smad1/5/8 protein were decreased,the cell viability was decreased and the number of migratory cells was increased(P<0.05).Compared with HH group,the expression ofα-SMA at mRNA and protein levels in rhBMP-7 group was decreased,the expression levels of CD31 mRNA and protein,Smad1/5/8 mRNA and p-Smad1/5/8 protein were increased,the cell viability was increased and the number of migratory cells was reduced(P<0.05).Compared with rhBMP-7 group,the expression ofα-SMA at mRNA and protein in DMH-1 group was increased,the expression levels of CD31 mRNA and protein,Smad1/5/8 mRNA and p-Smad1/5/8 protein were decreased,the cell viability was decreased and the number of migratory cells was increased(P<0.05).CONCLUSION:Hypoxia-hypercapnia conditions promote EndoMT in RPAECs,which promotes the development of hypoxia-hypercapnia pulmonary hypertension,and the underling mechanis

关 键 词：BMP-7/Smads通路 内皮-间充质转化 低氧高二氧化碳性肺动脉高压 肺动脉内皮细胞 

分 类 号：R363[医药卫生—病理学] R563[医药卫生—基础医学]