补元汤通过HIF-1/NOR-1途径改善低氧性肺血管重构的机制研究  

Study on the Mechanism of Buyuan Decoction Improving Hypoxic Pulmonary Vascular Remodeling Based on HIF-1/NOR-1 Pathway

作　　者：宋文龙 李瑾 Song Wenlong;Li Jin(Jiande Integrated traditional Chinese and Western Medicine Hospital,Zhejiang,Jiande 311600,China.)

机构地区：[1]浙江省建德市中西医结合医院,浙江建德311600

出　　处：《中国中医急症》2023年第10期1723-1727,1743,共6页Journal of Emergency in Traditional Chinese Medicine

基　　金：浙江省医药卫生科技计划(2020KY809)。

摘　　要：目的 通过观察补元汤方对缺氧条件下大鼠肺动脉平滑肌(PASMC)中低氧诱导因子(HIF-1)、孤儿核受体(NOR-1)、B细胞淋巴因子2(Bcl-2)、细胞抗凋亡蛋白2(c IAP2)、细胞周期蛋白D1(Cyclin D1)、Rb1等蛋白及mRNA表达的变化以及细胞活力、凋亡水平的影响,探讨其延缓肺动脉重构的可能机制。方法 将大鼠PASMC传代后置于不同氧浓度下培养,模拟慢性阻塞性肺疾病(COPD)所处低氧条件,予补元汤含药血清进行干预,WB检测HIF-1、NOR-1、Bcl-2、cIAP2、Rb1及Cyclin D1蛋白表达水平,并以RT-PCR检测上述m RNA表达,CCK-8、流式细胞分别检测细胞活力及凋亡水平变化。结果 经低氧培养后,10%氧浓度组大鼠PASMC细胞HIF-1、Cyclin D1、Bcl-2、cIAP2表达上升,细胞活力及凋亡率较常氧组上升,NOR-1、Rb1下调(P <0.05);当氧浓度下降至1%后,随着暴露时间的增加,PASMC细胞活力较常氧及中度低氧明显下降,凋亡率亦上升(P <0.05);电镜下大鼠肺动脉平滑肌细胞形态学发生改变;与两低氧组比较,经补元汤含药血清干预后,PASMC中HIF-1、Cyclin D1表达水平下调,NOR-1、Rb1表达上调,而Bcl-2、cIAP2表达抑制,细胞活力较常氧下降,且凋亡亦处于较低水平(P <0.05)。结论 补元汤对低氧性肺血管重构有保护作用,可能与抑制HIF-1、Cyclin D1表达,上调NOR-1、Rb1,抑制Bcl-2及cIAP2表达水平,降低PASMC细胞增殖活力,并使其凋亡亦处于较低水平有关。Objective:To discover the possible mechanism of Buyuan Decoction in delaying pulmonary artery remodeling through experimental observation on the expression of proteins and mRNA such as Bcl-2,Cyclin D1,cIAP2,HIF-1,NOR-1,Rb1,as well as cell viability and apoptosis levels in rat pulmonary artery smooth muscle(PASMC)under hypoxic condition.Methods:Rat PASMC was cultured under different oxygen concentrations to simulate the hypoxic conditions of chronic obstructive pulmonary disease(COPD).Buyuan Decoction containing serum was administered for intervention.The expression levels of Bcl-2,Cyclin D1,cIAP2,HIF-1,NOR-1,and Rb1 proteins were detected by WB,relative mRNA expression was detected by RT-PCR,and cell viability and apoptosis levels were measured by CCK-8 and flow cytometry,respectively.Results:After hypoxia cultivation,the expression of HIF-1,Cyclin D1,Bcl-2,and cIAP2 in PASMC cells of rats in the 10%oxygen concentration group increased,and the cell viability and apoptosis rate increased compared with the normoxic group.NOR-1 and Rb1 were down regulated(P<0.05).When the oxygen concentration decreased to 1%,as the exposure time increased,the PASMC cell viability significantly decreased compared with normoxic and moderate hypoxia,and the apoptosis rate also increased(P<0.05).Morphological changes of pulmonary artery smooth muscle cells in rats were observed under electron microscopy.Compared with the two hypoxic groups,after the intervention of Buyuan Decoc⁃tion containing serum,the expression levels of HIF-1 and Cyclin D1 were down regulated,while the expression of NOR-1 and Rb1 was upregulated,the expression of Bcl-2 and cIAP2 was down regulated.The activity of PASMC cells was lower than that of normoxia,and apoptosis was also at a lower level(P<0.05).Conclusion:Buyuan Decoction has a protective effect on hypoxic pulmonary vascular remodeling,which may be related to inhibiting the expression of HIF-1 and Cyclin D1,upregulating NOR-1 and Rb1,and inhibiting the expression levels of Bcl-2 and cIAP2,reducing the pro

关 键 词：肺动脉重构 低氧 肺动脉平滑肌细胞 补元汤 

分 类 号：R285.5[医药卫生—中药学]