紫菀酮和白藜芦醇在肺动脉高压细胞模型中的作用机制研究  

Study on the mechanism of shionone and resveratrol in pulmonary arterial hypertension cell model

作　　者：官占江 黑炳昌 郝文波 林树东 姜云飞 刘旭之 孟宪国 王媛媛 赵明宇 于海涛 杨磊 GUAN Zhan-jiang;HEI Bing-chang;HAO Wen-bo(Intensive Care Unit,The Third Affiliated Hospital of Qiqihar Medical College,Qiqihar 161000,China)

机构地区：[1]齐齐哈尔医学院附属第三医院重症医学科,161000 [2]齐齐哈尔医学院附属第三医院胸心外科,161000 [3]齐齐哈尔医学院附属第三医院临床检验,161000 [4]齐齐哈尔医学院附属第三医院呼吸内科,161000 [5]齐齐哈尔医学院附属第三医院临床药学,161000 [6]齐齐哈尔医学院附属第三医院血管外科,161000

出　　处：《中国现代药物应用》2025年第6期175-180,共6页Chinese Journal of Modern Drug Application

基　　金：2020年度黑龙江省省属本科高校基本科研业务费科学技术研究项目(项目编号:2020-KYYWF-00185),项目名称:白藜芦醇对实验性兔肺动脉高压相关炎症因子TNF-α以及IL-6表达的影响研究。

摘　　要：目的对比紫菀酮、白藜芦醇(RES)在肺动脉高压(PAH)细胞模型中的作用机制。方法体外培养大鼠肺动脉平滑肌细胞(PASMCs细胞)和大鼠肺动脉内皮细胞(rPAEC细胞),将PASMCs细胞和rPAEC细胞分别在含有2%O_(2)和5%CO_(2)的培养箱中培养24 h作为模型组,而将PASMCs细胞和rPAEC细胞正常条件下在含有21%O_(2)和5%CO_(2)培养箱中培养24 h作为对照组。分别使用含有2、4、8μg/ml紫菀酮和10、20、40μmol/L白藜芦醇处理模型组细胞,培养24 h,分别命名为紫菀酮低剂量组、紫菀酮中剂量组、紫菀酮高剂量组及白藜芦醇组低剂量组、白藜芦醇中剂量组、白藜芦醇高剂量组,每种细胞均分为8个组,每组均做6个重复。采用细胞增殖检测法(MTT法)检测各组不同细胞活力,流式细胞仪检测各组不同细胞凋亡水平,酶联免疫吸附试验(ELISA)检测各组不同细胞肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β和IL-6,逆转录实时定量聚合酶链式反应(RT-qPCR)和蛋白质印迹分析各组细胞内皮型一氧化氮合酶(eNOS)和沉默调节蛋白1(SIRT1)的mRNA相对表达量和蛋白表达水平。分析紫菀酮和白藜芦醇对PASMCs细胞和rPAEC细胞中eNOS和SIRT1的mRNA相对表达量和蛋白表达的影响,紫菀酮和白藜芦醇对PASMCs细胞和rPAEC细胞增殖和凋亡的影响,紫菀酮和白藜芦醇对PASMCs细胞和rPAEC细胞TNF-α、IL-1β、IL-6表达的影响。结果与对照组比较,模型组PASMCs细胞和rPAEC细胞的eNOS和SIRT1的mRNA相对表达量和蛋白表达水平均显著降低(P<0.05);与模型组比较,紫菀酮和白藜芦醇的低、中、高剂量组的PASMCs细胞和rPAEC细胞的eNOS和SIRT1的mRNA相对表达量和蛋白表达水平均显著提高(P<0.05);紫菀酮和白藜芦醇的中、高剂量组的PASMCs细胞和rPAEC细胞的eNOS和SIRT1的mRNA相对表达量和蛋白表达水平均显著高于低剂量组,紫菀酮和白藜芦醇的高剂量组的PASMCs细胞和rPAEC细胞的eNOS和SIRT1的mRNObjective To compare the mechanism of shionone and resveratrol(RES)in pulmonary arterial hypertension(PAH)cell model.Methods Rat pulmonary artery smooth muscle cells(PASMCs)and rat pulmonary artery endothelial cell(rPAEC)were cultured in vitro.PASMCs and rPAEC were cultured in an incubator containing 2%O_(2) and 5%CO_(2)for 24 h as a model group,while PASMCs and rPAEC were cultured in an incubator containing 21%O_(2) and 5%CO_(2)for 24 h under normal conditions as a control group.The cells in the model group were treated with 2,4,8μg/ml shionone and 10,20,40μmol/L resveratrol for 24 h,and were were named as low-dose shionone group,medium-dose shionone group,high-dose shionone group,low-dose resveratrol group,medium-dose resveratrol group,and high-dose resveratrol group,and each type of cells was divided into 8 groups,and 6 replicates were made in each group.Tetrazolium-based MTT assay was used to detect the viability of different cells in each group,flow cytometry was used to detect the apoptosis level of different cells in each group,enzyme-linked immunosorbent assay(ELISA)was used to detect the tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6 of different cells in each group,and reverse transcription real-time quantitative polymerase chain reaction(RT-qPCR)and western blot were used to analyze the relative mRNA expression levels and protein expression levels of endothelial nitric oxide synthase(eNOS)and silent mating type information regulation 2 homolog 1(SIRT1)in each group.Analysis of the effects of shionone and resveratrol on the relative mRNA expression and protein expression of eNOS and SIRT1 in PASMCs and rPAEC,the effects of shionone and resveratrol on proliferation and apoptosis of PASMCs and rPAEC,and the effects of shionone and resveratrol on expression of TNF-α,IL-1βand IL-6 in PASMCs and rPAEC.Results The relative mRNA expression and protein expression levels of eNOS and SIRT1 in PASMCs and rPAEC in the model group were significantly reduced compared with the control group(P<0.05).C

关 键 词：紫菀酮 白藜芦醇 肺动脉高压 沉默调节蛋白1 肺动脉平滑肌细胞 内皮细胞 

分 类 号：R285.5[医药卫生—中药学]