人参皂苷Rg1和Rb1对流感病毒感染细胞的影响及其机制研究  

Effect and mechanism of ginsenoside Rgl and Rbl on influenza A virus infection

作　　者：张春晶[1,2] 顾立刚[2] 牛旭艳[2] 王甫[2] 彭桂英[2] 

机构地区：[1]北京中医药大学基础医学院中西医结合基础中医药防治病毒性疾病教育部重点实验室,100029 [2]齐齐哈尔医学院生化与分子生物学教研室

出　　处：《中华微生物学和免疫学杂志》2009年第10期894-898,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30772872）;国家自然科学基金青年科学基金资助项目（30701105）

摘　　要：目的研究人参皂苷Rg1和Rb1对流感病毒感染所致的细胞氧化应激损伤、细胞内转录因子（NF—KB和AP—1）转录活性及前炎症细胞因子IL-8释放的影响。方法甲型流感病毒H3N2作为刺激因素，以DCFH—DA为探针，流式细胞仪检测细胞内活性氧（iROS）产生水平。利用双荧光素酶顺式报告系统，榆测人参皂苷Rg1和Rb1对NF—κB—luc及AP-1-luc相对荧光素酶值的影响。RT—PCR法进一步验证人参皂苷Rg1和Rb1对TL-8mRNA表达水平的影响。结果病毒感染后细胞中ROS产生增加45％～55％，而给予有效浓度人参皂苷Rg1和Rb1后，荧光强度降低，与正常细胞内自由基含量基本相同。通过NF-κB及AP-1双荧光素酶报告系统检测发现，与正常对照组比较，病毒感染的细胞中NF—κB及AP-1荧光素酶报告活性明显升高；与病毒损伤组比较，人参皂苷Rg1、Rb1治疗组NF—κB及AP—1荧光素酶报告活性明显受到抑制。RT—PCR结果显示人参皂苷Rg1和Rh1能明显抑制病毒诱导的1L-8mRNA表达水平的变化，使之趋近于正常对照组水平。结论人参皂苷Rg1和Rb1可拈抗病毒感染细胞中ROS产生水平、降低氧化应激敏感的信号传导通路Nn-κB及AP-1的转录活性，并抑制二者下游靶基因IL-8表达。Objective To study the effect of ginsenoside Rg1 and Rb1 on the oxygen radical scavenger, antioxidant and antiinflammatory agent through inhibition nuclear factor-kappa B（NF-KB） and activator protein-1 （ AP-1 ） activation, as well as IL-8 release infected by influenza virus. Methods Epithelial cells was exposed to influenza virus infection for 2 h before treatment with Rgland Rbl for 24 h. The quantities of generation of intracellular reactive oxygen species（iROS） was analyzed by flow cytometric analysis represented by DCF fuorescence. NF-κB-responsive elemcnt-luciferasc reportor genc and AP-1-responsive element-luciferase reporter genc was transfected and dual-luciferase cis-reporting systems was used to assay the transcriptional activity of NF-κB and AP-1 under the stimulated circumstance of influenza virus infection. RT-PCR was used to detect the gene transcription level of IL-8. Results Oxidant production, measured as the fluorescence of dichlorofluorescein, was significant increased of 45% -55% by influenza virus infection. This increase in iROS generation was significant blocked by Rg1 and Rb1. The relative luciferase reporter assay of NF-κB or AP-1 is apparently improved by influenza virus infection. But ginsenoside Rgl and Rbl significantly repressed the relative value of luciferase. RT-PCR showed upregulation of inflammatory markers IL-8 in A549 cells infected by influenza virus, Rgl and Rbl had a significant effect on the expression of IL- 8 after treated with influenza virus. Conclusion Ginsenoside Rgland Rbl play a protective role by scavenging of reactive oxygen species（ROS） , reducing transcriptional activation of NF-κB and AP-1 , as well as on the downregulation of the downstream inflammatory markers target gene expression IL-8.

关 键 词：人参皂苷Rg1和Rb1 甲型H3N2流感病毒 氧化应激 核因子-ΚB 激活蛋白-1 

分 类 号：R28[医药卫生—中药学]