流感病毒性肺炎小鼠肺组织炎性细胞因子表达及疏风宣肺方和解表清里方的调控作用  

The expression of the inflammation-related cytokines in pneumonia mice infected with influenza virus and regulation of Shufengxuanfei and Jiebiaoqingli herbal anti-virus formulas

作　　者：卢娜娜[1] 刘琪[1,2] 顾立刚[1] 吴珺[1] 邱泽计[1] 张洪春[3] 晁恩祥[3] 张沂[1] 于卓男[1] 

机构地区：[1]北京中医药大学基础医学院中医药抗病毒实验室 [2]山西中医学院,山西太原030024 [3]北京中日友好医院

出　　处：《细胞与分子免疫学杂志》2014年第3期254-257,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81173371);北京中医药大学自主课题(2013-JYBZZ-XS-016)

摘　　要：目的研究流感病毒性肺炎小鼠肺组织炎性细胞因子表达及疏风宣肺方和解表清里方的调控作用。方法制备甲型流感病毒性肺炎小鼠模型,随机分为空白组、肺炎模型组、奥司他韦对照组、疏风宣肺方大、中、小剂量组(SL、SM和SS),解表清里方大、中、小剂量组(JL、JM和JS)。提取总RNA,采用基因芯片Pathway分析,挑选参与调控炎症相关通路中的靶基因。同时,通过实时荧光定量PCR(qRT-PCR)对IL-1β、IL-8、IL-10、CCL5(RANTES)、ICAM-1的mRNA表达水平进行验证。采用Western blot法对肺组织IL-1β的表达进行验证。结果模型组差异表达基因IL-1β、CXCR2、CCL5、IL-10、IL-6、IL-18、TGF-β1、CCL2明显上调,较正常组差异显著。各剂量治疗组对IL-1β、CXCR2、IL-10、IL-6表达有显著的下调作用,SM及SS组下调IL-18、TGF-β1、CCL2、CCL5基因的表达,JM及JL组下调差异表达基因IL-18、CCL5。qRT-PCR和Western blot法结果显示,两种方药各剂量组均可降低IL-1β的mRNA与蛋白表达(P<0.05或P<0.01)。SM组和JM组对IL-8、IL-10、RANTES、ICAM-1的mRNA有显著抑制(P<0.05或P<0.01)。qRT-PCR结果与基因芯片结果基本一致。结论疏风宣肺方和解表清里方均可抑制流感病毒感染后肺组织的免疫炎症损伤。Objective To investigate the expression of the inflammation-related cytokines in pneumonia mice infected with influenza virus and regulation of Shufengxuanfei（SFXF） and Jiebiaoqingli （JBQL） Chinese herbal anti-virus formulas. Methods Mice were anesthetized and then infected intranasally by dropping 0.05 mL of influenza virus suspension （4 x LDso） except normal group. Mice were divided randomly into nine groups： normal group, model group （virus only）, control group r ll. 375 g./（kg · d）Oseltamivir], low-dose SFXF [0.94 g/（ kg · d）], medium-dose SFXF [ 1.88 g/（ kg · d）], high-dose SFXF [3.76 g/（ kg · d） ], low-dose JBQL [ 1.09 g/（ kg · d） ], medium-dose JBQL [2.18 g/（ kg · d） ] and high-dose JBQL [4.36 g/（kg · d）]. Oseltamivir group, SFXF groups and JBQL groups were administered to mice by oral gavage in equal dose of 0.2 mL daily for 4 consecutive days, while the rest of the groups received water only. Total RNA was extracted in each group. Then gene chips were used to screen these RNA samples. Select differentially expressed genes of cytokines involved in inflammation. Some candidate genes, such as IL-1β, IL-8, IL-10, RANTES and ICAM-I were verified by qRT-PCR. To confirm the genes expression data from the microarray involved in inflammation in response to virus infection and treatment, we used qPCR to verify mRNA relative expressions of IL-β3, IL-8, IL-10, RANTES and ICAM-1. The expression of IL-1β protein in lung tissues was verified by Western blotting. Results IL-115, CXCR2, CCL5, IL-10, IL-6, IL-18, TGF-β1 and CCL2 were up-regulated in model group. Gene expressions of IL-1β, CXCR2, CCLS, IL-10 and IL-6 were significantly down-regulated by all therapeutic groups. SFXF in medium-dose and low-dose down-regulated gene expressions of IL-18, TGF-β, CCL2 and CCL IL-18 and CCL5 was down-regulated by both low-dose and medium-dose JBQL. qRT-PCR and western blot experiments showed that two formulas in medium-dose can down-regulate mRNA and protein expression of

关 键 词：疏风宣肺方 解表清里方 肺炎 流感病毒 炎症 细胞因子 

分 类 号：R563.1[医药卫生—呼吸系统] R392.11[医药卫生—内科学]