疏风宣肺方、解表清里方药对流感病毒小鼠TLR7、MyD88、NF-κB mRNA及蛋白表达的影响

Effect of Shufeng Xuanfei Recipe and Jiebiao Qingli Recipe on mRNA and Protein Expressions of TLR7， MyD88， and NF-κB in Mice Infected with Influenza Virus

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DOI：10.7661/CJIM.2013.09.1256

中文关键词: 疏风宣肺方解表清里方流感病毒 Toll样受体7

英文关键词:Shufeng Xuanfei Recipe Jiebiao Qingli Recipe influenza virus Toll-like receptor 7

基金项目:国家自然科学基金资助项目（No81173371）

作者	单位	E-mail
刘琪,卢娜娜,周旭澎
顾立刚	北京中医药大学基础医学院中医药防治病毒性疾病实验室（北京 100029）	lggulg@163.com

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中文摘要:

目的观察疏风宣肺和解表清里方药对流感病毒亚甲型肺适应株（FM1）感染小鼠肺组织细胞Toll样受体7（Toll-like receptor 7， TLR7）、髓样分化因子88（myeloid differentiation factor 88， MyD88）、激活核因子Kappa B（nuclear factor-kappaB，NF-κB） mRNA及蛋白表达的影响。方法选择108只小鼠，随机分为9组：正常组，模型组，磷酸奥司他韦胶囊组［西药组，2.5 g/（mL?d）］，疏风宣肺方（中药1）高、中、低剂量组［3.762、1.881、0.941 g/（kg?d）］，解表清里方（中药2）高、中、低剂量组［4.368、2.184、1.092 g/（kg?d）］，每组12只。将各组小鼠用乙醚轻度麻醉，正常组以0.05 mL生理盐水滴鼻，其余各组以0.05 mL 4LD50流感病毒FM1稀释液滴鼻感染小鼠。造模成功率100%。感染后2 h灌胃给药。正常组、模型组灌胃蒸馏水， 0.2 mL/次，1次/d，连续干预4天。采用RT-PCR反应测定肺组织TLR7、MyD88、NF-κB mRNA表达，采用Western blot法测定肺组织TLR7、MyD88、NF-κB蛋白表达水平。结果与正常组比较，模型组TLR7、MyD88、NF-κB mRNA表达升高（P<0.01）；与模型组比较，西药组，中药1高、中、低剂量组TLR7、MyD88、NF-κB mRNA及蛋白表达降低（P<0.05，P<0.01），中药2高、中剂量组TLR7、NF-κB mRNA及蛋白表达降低（P<0.05，P<0.01），中药2高、中剂量组MyD88 mRNA表达降低（P<0.05），中药2中剂量组MyD88蛋白表达降低（P<0.05），中药2低剂量组TLR7、NF-κB蛋白表达降低（P<0.05）。与西药组比较，中药1低剂量组MyD88蛋白表达降低（P<0.05），中药1中、低剂量组NF-κB蛋白表达降低（P<0.01）；中药2高、中、低剂量组TLR7 mRNA及蛋白表达降低（P<0.05，P<0.01），MyD88蛋白表达降低（P<0.01），中药2低剂量组NF-κB mRNA及蛋白表达降低（P<0.05，P<0.01）。结论疏风宣肺方各剂量和解表清里中剂量能通过调节MyD88依赖的TLR信号传导通路下调NF-κB活性，发挥抗流感病毒的作用。疏风宣肺方效果优于解表清里方。

英文摘要:

Objective To observe effect of Shufeng Xuanfei Recipe （SXR） and Jiebiao Qingli Recipe （JQR） on mRNA and protein expressions of Toll-like receptor 7 （TLR7）， myeloid differentiation factor88 （MyD88）， and nuclear factor-kappaB （NF-κB） in mice infected with influenza virus FM1. Methods One hundred and eight mice were randomly divided into nine groups， i.e.， the normal control group， the model group， the Oseltamivir group （at the daily dose of 2.5 g/mL）， the high dose SXR group （at the daily dose of 3.762 g/kg）， the middle dose SXR group （at the daily dose of 1.881 g/kg）， the low dose SXR group （at the daily dose of 0.941 g/kg）， the high dose JQR group （at the daily dose of 4.368 g/kg），the middle dose JQR group （at the daily dose of 2.184 g/kg）， and the low dose JQR group （at the daily dose of 1.092 g/kg）， 12 in each group. All mice were mildly anesthetized by ether. Mice in the normal control group were treated by nasal drop of 0.05 mL normal saline， while mice in the rest groups were infected by nasal drop of 0.05 mL influenza virus strain FM1 （LD50）. The successful modeling rate was 100%. All medication was performed by gastrogavage 2 h after infection. Distilled water was given by gastrogavage to mice in the normal control group and the model group at the daily dose of 0.2 mL， each time per day for 4 successive days. mRNA expressions of TLR7， MyD88， and NF-κB in the lung tissue were determined by Western blot. Results Compared with the normal control group， mRNA expressions of TLR7， MyD88， and NF-κB increased in the model group （P<0.01）. Compared with the model group， mRNA and protein expressions of TLR7， MyD88， and NF-κB decreased in the Oseltamivir group， the high， middle， and low dose SXR groups （P<0.05，P<0.01）； mRNA and protein expressions of TLR7 and NF-κB decreased in the high and middle dose JQR groups （P<0.05，P<0.01）； mRNA expressions of MyD88 decreased in the high and middle dose JQR groups （P<0.05）； protein expressions of MyD88 decreased in the middle dose JQR group （P<0.05）； protein expnessions of TLR7 and NF-κB decreased in the low dose JQR group （P<0.05）. Compared with the Oseltamivir group， protein expressions of MyD88 decreased in the low dose SXR group （P<0.05）； protein expressions of NF-κB decreased in the middle and low dose SXR groups （P<0.01）； mRNA and protein expressions of TLR7 （P<0.05，P<0.01）， and protein expressions of MyD88 （P<0.01） decreased in the high， middle， and low dose JQR groups； mRNA and protein expressions of NF-κB decreased in the low dose JQR group （P<0.05，P<0.01）. Conclusions Each dose SXR and middle dose JQR could down-regulating the activity of NF-κB through adjusting MyD88 dependent TLR signal pathway， thus fighting against influenza virus. SXR was more effective than JQR.

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