大黄对呼吸道合胞病毒抑制作用的研究  

Inhibitory effect of Rhubarb on respiratory syncytial virus

作　　者：陈美汐 申淋潮 吴欣 王自辉[4] 宋雪倩 郝智仪 王秀丽[3] Chen Meixi;Shen Linchao;Wu Xin;Wang Zihui;Song Xueqian;Hao Zhiyi;Wang Xiuli(Graduate School of Hebei Medical University,Shijiazhuang 050017,China;Xi′an Center for Disease Control and Prevention,Xi′an 710054,China;Department of Clinical Laboratory,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Traditional Chinese Medicine,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;College of Laboratory Technology,Hebei Medical University,Shijiazhuang 050017,China)

机构地区：[1]河北医科大学研究生院,石家庄050017 [2]西安市疾病预防控制中心,710054 [3]河北医科大学第二医院检验科,石家庄050000 [4]河北医科大学第二医院中医外科,石家庄050000 [5]河北医科大学检验技术学院,石家庄050017

出　　处：《国际呼吸杂志》2024年第4期459-466,共8页International Journal of Respiration

基　　金：河北省中医药管理局科研计划项目(2021142)。

摘　　要：目的探究在A549细胞中大黄对呼吸道合胞病毒复制的影响及在感染后调节自噬的作用。方法本研究为实验研究,采用呼吸道合胞病毒Long株感染人非小细胞肺癌系A549细胞,构建呼吸道合胞病毒感染细胞模型。使用A549细胞设置感染对照组、利巴韦林组、大黄低剂量组、大黄中剂量组、大黄高剂量组,在感染呼吸道合胞病毒和大黄处理24 h后,使用免疫印迹法检测细胞内病毒荧光蛋白(RSV-GFP)、微管相关蛋白1A/1B-轻链3(LC3)蛋白表达的相对水平,定量聚合酶链反应检测细胞内病毒基因(RSV-N)基因的相对量的表达。在感染呼吸道合胞病毒和大黄处理72 h后,使用CCK8法测定细胞活性,病毒空斑实验法测定上清病毒滴度。设置入胞感染对照组、入胞大黄组;吸附感染对照组、吸附大黄组;灭活感染对照组、灭活大黄组,在感染呼吸道合胞病毒和大黄处理24 h后,使用定量聚合酶链反应检测细胞内病毒基因(RSV-N)基因的相对量的表达,在感染呼吸道合胞病毒和大黄处理72 h后,使用CCK8法测定细胞活性。结果(1)不同浓度的大黄培养A549细胞72 h后存活率比较有统计学意义(P<0.001),与5 mg/L比较,500、1000 mg/L均有统计学意义(均P<0.05),10、25、50、100 mg/L均无统计学意义(均P>0.05),不同浓度利巴韦林培养A549细胞72 h后存活率比较无统计学意义(F=1.02,P=0.442);结果显示对于A549细胞100 mg/L大黄为最高安全剂量,本实验选取25、50、100 mg/L分别作为大黄低剂量组、大黄中剂量组、大黄高剂量组处理浓度,利巴韦林处理浓度为500μmol/L。(2)大黄低剂量组细胞存活率较感染对照组[(81.43±1.51)%比(37.07±1.59)%]升高、大黄中剂量组细胞存活率较大黄低剂量组[(89.06±1.23)%比(81.43±1.51)%]升高、大黄高剂量组细胞存活率(99.84±1.45)%、利巴韦林组(97.41±1.60)%较大黄中剂量组(89.06±1.23)%升高(均P<0.05)。(3)5组细胞RSV-N基因表达量及上�Objective To investigate the effect of Rhubarb on respiratory syncytial virus replication in A549 cells and its role in regulating autophagy after infection.Methods This was an experimental study.Human non-small cell lung cancer(NSCLC)A549 cells were infected with respiratory syncytial virus(RSV)Long strain to establish an RSV infected cell model.A549 cells were used to set up infection control group,ribavirin group,Rhubarb lowdose group,Rhubarb mediumdose group,and Rhubarb highdose group.After being infected with RSV and treated with Rhubarb for 24 h,western blot was used to detect the relative expression level of intracellular viral fluorescent protein(RSV-GFP)and autophagy marker(LC3)protein,and quantitative polymerase chain reaction was used to detect the relative expression levels of intracellular viral gene(RSV-N).72 h after infection with respiratory syncytial virus and treatment with Rhubarb,cck8 assay was used to determine the cell activity and viral plaque assay was employed to determine the viral titer of supernatant.The following groups-control group and Rhubarb group,adsorbed infection control group and adsorbed Rhubarb group,inactivated control group and inactivated Rhubarb group-were all set up.The relative expression of intracellular viral gene(RSV-N)was detected by quantitative polymerase chain reaction after infection with RSV and Rhubarb treatment for 24 h.The cellular activity was detected by cck8 method 72 h after infection with RSV and Rhubarb treatment.Results(1)The survival rate of A549 cells cultured with different concentrations of Rhubarb after 72 hours was statistically significant(P<0.001).Compared with 5 mg/L,500 and 1000 mg/L groups showed statistical significance(all P<0.05);10,25,50 and 100 mg/L groups showed no statistical significance(all P>0.05).There was no statistical significance in survival rate of A549 cells cultured with different concentrations of ribavirin for 72 h(F=1.02,P=0.442).For A549 cells,100 mg/L Rhubarb was the highest safe dose,and 25,50,and 100 mg/L were sele

关 键 词：呼吸道合胞体病毒感染 大黄 抑制作用 A549细胞 

分 类 号：R285[医药卫生—中药学]