CAJ | 학술논문

目的观察疏风宣肺和解表清里方药对流感病毒性肺炎小鼠肺组织CD4+辅助性T细胞1/2(Th1/2)平衡的调控作用.方法选择SPF级雄性ICR小鼠108只,按随机数字表法分为对照组、模型组、达菲组以及疏风宣肺方高、中、低剂量组和解表清里方高、中、低剂量组9组,每组12只.采用流感病毒亚甲型鼠肺适应株(FM1)滴鼻感染小鼠建立流感病毒性肺炎模型,对照组以0.05 ml生理盐水滴鼻.于病毒感染后2 h,对照组、模型组灌胃蒸馏水;达菲组给予达菲2.5 g·ml-1·d-1;其余6组分别灌服疏风宣肺抗流感方药(主要药物金银花、连翘、牛蒡子、蝉蜕、荆芥等)高、中、低剂量(3.762、1.881、0.941 g·kg-1·d-1)和解表清里抗流感方药(主要药物炙麻黄、生石膏、黄芩、杏仁、生甘草等)高、中、低剂量(4.368、2.184、1.092 g·kg-1·d-1);均每日1次,每次0.2 ml,连用4 d.采用实时荧光定量聚合酶链反应(PCR)和蛋白质免疫印迹法(Western blotting)分别测定肺组织γ-干扰素(IFN-γ)、白细胞介素-4(IL-4)mRNA以及蛋白表达水平.结果与对照组比较,模型组IFN-γmRNA及蛋白表达均明显降低,IL-4 mRNA及蛋白表达均明显升高(均P<0.05).与模型组比较,各药物组IFN-γmRNA及蛋白表达均升高,以疏风宣肺方低剂量组和解表清里方中剂量组升高更显著(IFN-γmRNA:0.85±0.14、0.58±0.06比0.15±0.48,均P<0.01);IL-4 mRNA及蛋白表达均有所降低,以疏风宣肺方中剂量组降低更显著(IL-4 mRNA:1.06±0.19比2.89±0.75,P<0.05).结论疏风宣肺方通过升高IFN-γ水平、降低IL-4水平,纠正Th1/2的平衡,减轻肺组织免疫病理损伤,以中低剂量组为优.
목적관찰소풍선폐화해표청리방약대류감병독성폐염소서폐조직CD4+보조성T세포1/2(Th1/2)평형적조공작용.방법선택SPF급웅성ICR소서108지,안수궤수자표법분위대조조、모형조、체비조이급소풍선폐방고、중、저제량조화해표청리방고、중、저제량조9조,매조12지.채용류감병독아갑형서폐괄응주(FM1)적비감염소서건립류감병독성폐염모형,대조조이0.05 ml생리염수적비.우병독감염후2 h,대조조、모형조관위증류수;체비조급여체비2.5 g·ml-1·d-1;기여6조분별관복소풍선폐항류감방약(주요약물금은화、련교、우방자、선세、형개등)고、중、저제량(3.762、1.881、0.941 g·kg-1·d-1)화해표청리항류감방약(주요약물자마황、생석고、황금、행인、생감초등)고、중、저제량(4.368、2.184、1.092 g·kg-1·d-1);균매일1차,매차0.2 ml,련용4 d.채용실시형광정량취합매련반응(PCR)화단백질면역인적법(Western blotting)분별측정폐조직γ-간우소(IFN-γ)、백세포개소-4(IL-4)mRNA이급단백표체수평.결과여대조조비교,모형조IFN-γmRNA급단백표체균명현강저,IL-4 mRNA급단백표체균명현승고(균P<0.05).여모형조비교,각약물조IFN-γmRNA급단백표체균승고,이소풍선폐방저제량조화해표청리방중제량조승고경현저(IFN-γmRNA:0.85±0.14、0.58±0.06비0.15±0.48,균P<0.01);IL-4 mRNA급단백표체균유소강저,이소풍선폐방중제량조강저경현저(IL-4 mRNA:1.06±0.19비2.89±0.75,P<0.05).결론소풍선폐방통과승고IFN-γ수평、강저IL-4수평,규정Th1/2적평형,감경폐조직면역병리손상,이중저제량조위우.
Objective To investigate regulatory effects of Shufeng Xuanfei and Jiebiao Qingli Formulae on CD4+helper T cell 1/2(Th1/2)cytokines of mice with influenza virus FM1 pneumonia. Methods One hundred and eight male mice,ICR grade,were randomly divided into nine groups:control group,model group,oseltamivir group, the high,middle and low dose groups of Shufeng Xuanfei Formula and the high,middle and low dose groups of Jiebiao Qingli Formula(each,n=12). The pneumonia model was established by dropping influenza virus strain FM1 into the nose,while the control group was given normal saline 0.05 ml into the nose. After the above infectious treatment of mice for 2 hours,they were treated with oseltamivir 2.5 g·ml-1 ·d-1 for 4 days in oseltamivir group, high,middle and low doses(3.762,1.881,0.941 g·kg-1 ·d-1)of Shufeng Xuanfei Formula(the main ingredients including honeysuckle, forsythia,burdock,periostraum cicada,schizonepeta,et al.)and high,middle and low dose(4.368,2.184, 1.092 g·kg-1 ·d-1)of Jiebiao Qingli Formula〔mainly ephedra,gypsum,scutellaria baicalensis,apricot(almond),raw glycyrrhiza,et al.〕were administered intragastrically in the corresponding groups,0.2 ml once a day for 4 days;in the control and model groups,the mice were treated by intragastrical administration of the same amount of normal saline for 4 days. The mRNA and protein expressions of interferon-γ(IFN-γ)and interleukin-4(IL-4)were determined by real-time fluorescent quantitative polymerase chain reaction(PCR)and Western blotting. Results Compared with control group,in model group,the expressions of IFN-γmRNA and protein were significantly down-regulated,while those of IL-4 mRNA and protein,obviously up-regulated(all P<0.05). Compared with model group,in all the drug treatment groups,the expressions of IFN-γmRNA and protein were significantly up-regulated,especially prominent in the low dose group of Shufeng Xuanfei Formula and the middle dose group of Jiebiao Qingli Formula(IFN-γmRNA:0.85±0.14,0.58±0.06 vs. 0.15±0.48,both P<0.01),while those of IL-4 mRNA and protein,obviously down-regulated,especially marked being in the middle dose group of Shufeng Xuanfei Formula(IL-4 mRNA:1.06±0.19 vs. 2.89±0.75,P<0.05). Conclusion The middle and low dose of Shufeng Xuanfei Formula can ameliorate the immune-pathological injury of influenza viral pneumonia by regulating the balance of Th1/Th2 through up-regulating the expression of IFN-γand down-regulating the expression of IL-4.