目的:观察疏风宣肺解毒方对流感病毒性肺炎小鼠肺组织Janus激酶信号转导与转录激活因子(JAK-STAT)通路的调控作用。方法将60只小鼠按随机数字表法分为正常组、模型组、达菲对照组和疏风宣肺方高、中、低剂量组,每组10只。应用0.05 mL的4LD50流感病毒肺适应株FM1滴鼻感染小鼠,建立小鼠流感病毒性肺炎模型;正常组以0.05 mL生理盐水滴鼻。制模成功2 h后,正常组、模型组灌服蒸馏水;达菲对照组灌服达菲(磷酸奥司他韦)2.5 g·mL-1·d-1;疏风宣肺方高、中、低剂量组分别灌服疏风宣肺解毒方药(由菊花、桑叶、杏仁、桔梗、连翘、柴胡等组成,颗粒剂),按照人与小鼠体表面积换算给药剂量,以加倍量为高剂量,折半量为低剂量,每日1次,每次0.2 mL。连续给药4 d后取小鼠肺组织,采用基因芯片技术检测小鼠JAK-STAT通路相关差异基因的表达,筛选差异表达基因的标准为:上调基因P<0.05,且log2比值>1,下调基因P<0.05,且log2比值<-1。应用实时荧光定量反转录-聚合酶链反应(RT-qPCR)测定肺组织Janus激酶(JAK)、γ干扰素(IFN-γ)的mRNA表达水平。结果与正常组比较,模型组差异表达基因STAT5〔log2(正常组/模型组)=2.32〕、白细胞介素-4受体亚单位〔IL4RA,log2(正常组/模型组)=4.77〕、白细胞介素-12受体〔IL12R, log2(正常组/模型组)=1.58〕、 JAK〔log2(正常组/模型组)=2.41〕均明显上调,干扰素(IFN)明显下调〔log2(正常组/模型组)=-1.45〕;与模型组比较,达菲对照组〔log2(达菲对照组/模型组)=1.51〕、方药各组〔log2(方药低剂量组/模型组)=1.46,log2(方药中剂量组/模型组)=1.72,log2(方药高剂量组/模型组)=1.40〕差异表达基因IFN明显上调,STAT5〔log2(达菲对照组/模型组)=-2.06,log2(方药低剂量组/模型组)?
ObjectiveTo investigate the regulatory effects of traditional Chinese medicine (TCM) Shufengxuanfeijiedu formula on Janus kinase signal transducer and activators of transcription (JAK-STAT) of lung tissues in mice with influenza viral pneumonia.Methods According to random number table, 60 mice were randomly divided into six groups with 10 mice in each group: normal group (N), model group (M), Tamiflu control group (C) and low (SL), medium (SM), high dose (SH) Shufengxuanfeijiedu formula groups. The mouse model of influenza virus pneumonia was reproduced by dropping of 0.05 mL 4LD50 inflluenza virus FM1 strain which can be adapted to lung tissue into the nose; while the N received nose instillation of 0.05 mL normal saline. After successful modeling for 2 hours, distilled water was given orally (by lavage) to N and M; Duffy (oseltamivir) 2.5 g·mL-1·d-1 was administrated to C; the TCM SL, SM, SH were intragastrically administered with different doses of shufengxuanfeijiedu decoction into the corresponding groups respectively (the ingredients of prescription: chrysanthemum, mulberry leaf, almond, platycodon root, forsythia, bupleurum etc. forming granules), according to the suitable dose of granules used for human body surface, the dose used for mouse surface area was calculated, the high dose means the dose used in the medium dose group doubled, the low dose means 1/2 dose used in medium group, once a day, once 0.2 mL for consecutive 4 days. Afterwards, the lung tissues were collected, the mouse differential gene expressions related to JAK-STAT pathway were detected by gene chip technology, the standards for screening of differential gene expression were as follows: up-regulated gene was P 1; down-regulation gene wasP 〈 0.05, and log2ratio 〈 -1. The levels in lung tissue kinase (JAK) andγinterferon (IFN-γ) mRNA expressions were determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR).Results Com