Based On The MAPK Signal Transduction Pathway To Explore Clear Lung Low School Particles On The Immune Effects Of Bacterial Lung Infections BALB/C Mice

Objective:Through the establishment of immune compromised mouse model of lunginfection, discuss Qing-Fei-Pei-Yuan granules affect clearing the lungs of MAPKsignaling pathway protein p38,JNK, ERK protein and mRNA expression, therebyinvestigating the inflammatory process of improving immune function and the controlmechanism for clinical Qing-Fei-Pei-Yuan granules provide experimental evidence fora wide range of applicationsMethods:1. Animal grouping: The frist grouping: Sixty female BALB/c mouse wereaveraged divided randomly into two groups according to the random number table:normal group(12rats in each group), model group(48rats in each group), After thesuccess of modeling, due to anesthesia death4, the remaining44mice. The secondgrouping: forty-four female BALB/c mouse were averaged divided randomly into fourgroups according to the random number table: model group,AZT group,Tang Herbgroup, Qing-Fei-Pei-Yuan granules group,11rats in each group, in order todistinguish we dyed them with Picric Acid solution.2. Animal model making and Administration: The normal group was given0.9%saline according0.25ml/10g standards by intrapersonal injection after three days,the model group vaccinate an equal volume of FLV virus. According to FLV viruspathogenic characteristics, determine the modeling period was21d. When immunecompromised mouse model is successful, injected0.03-0.05ml10%chloral hydrateinto abdominal, then gave them50μl the Ⅲtype pneumococcal bacterial suspensionusing intranasal method, According to pathogenic characteristics of the ⅢtypeStreptococcus pneumonia, determine the modeling period is7days. After one dayfrom successful modeling, then begin to fed mice of each group with correspondingdrug, the AZT group was given AZT suspension（0.08g/kg. d）for14days, Tang Herbgroup was given Tang Herb tablet suspension （2.56g/kg.d） for14days,Qing-Fei-Pei-Yuan granules group was given Qingfeipeiyuan particles（2g/kg.d）for 14days, the normal group and model group were given0.9%saline for14days, Thefrequency of drenching of all groups are one time per day, and dose is0.2ml10g-1.3. Index detection: Observe the general condition of the mice and measure bodyweight and abdominal circumference dynamically in the experiment. Put the mice todeath6hours after the last administration, remove the lung and divide it into two partsaseptically, then mark and store them in vials, preserve them in-80℃refrigerator.One part of these lung tissue were measured the p38、JNK、ERK protein expression byWestern Blot method, the other part were measured the p38、JNK、ERK mRNAexpression by Real time PCR method.Results:1. The general condition of mice were improved in different degrees in eachtreatment group Comparison with the model group compared with the model group.2. p38、 JNK、 ERK mRNA and protein expression of the lung tissue aresignificantly higher than the model group(P <0.05),After drug intervention,Qing-Fei-Pei-Yuan granules group and other groups are reduced to different extents.3. p38protein expression of the lung tissue in Qing-Fei-Pei-Yuan granules groupand other groups are lower than the mode group, and the difference was statisticallysignificant (P <0.05);It is also lower than Tang Herb group, the difference arestatistically significant (P <0.05);It has no difference between Qing-Fei-Pei-Yuangranules group and AZT group(P＞0.05).4. JNK protein expression of the lung tissue in Qing-Fei-Pei-Yuan granules groupand other groups are lower than the model group, and the difference was statisticallysignificant (P <0.05);It is lower than AZT group and Tang Herb group, the differenceare statistically significant (P <0.05).5. ERK protein expression of the lung tissue in Qing-Fei-Pei-Yuan granulesgroup and AZT group and Tang Herb group are lower than the model group, and thedifference was statistically significant (P <0.05); ERK protein expression of the lungtissue in Qing-Fei-Pei-Yuan granules group is lower than the AZT group, and thedifference was statistically significant (P <0.05); it has no difference betweenQing-Fei-Pei-Yuan granules group and Tang Herb group(P＞0.05).6. When Qing-Fei-Pei-Yuan granules group and AZT group and Tang Herb groupcompared with the model group,2-△△ct＜0.5，It explains that p38mRNA expression of Qing-Fei-Pei-Yuan granules group and AZT group and Tang Herb group wassignificantly reduced compared with the model group; Qing-Fei-Pei-Yuan granulesgroup compared with the Tang Herb group,2-△△ct＞2，It explains that p38mRNAexpression of Qing-Fei-Pei-Yuan granules group was significantly increasedcompared with the Tang Herb group; Qing-Fei-Pei-Yuan granules group comparedwith AZT group,0.5＜2-△△ct＜2，It explains that p38mRNA expression of these twogroups has no difference.7.①When Qing-Fei-Pei-Yuan granules group and AZT group and Tang Herbgroup compared with the model group,2-△△ct＜0.5，It explains that JNK1mRNAexpression of Qing-Fei-Pei-Yuan granules group and AZT and Tang Herb group wassignificantly reduced compared with the model group; Qing-Fei-Pei-Yuan granulesgroup compared with the AZT group and Tang Herb group,2-△△ct＜0.5，It explains thatJNK1mRNA expression of Qing-Fei-Pei-Yuan granules group was significantlyreduced compared with the AZT group and Tang Herb.②When Qing-Fei-Pei-Yuangranules group and AZT group and Tang Herb group compared with the model group,2-△△ct＜0.5，It explains that JNK2mRNA expression of Qing-Fei-Pei-Yuan granulesgroup and AZT and Tang Herb group was significantly reduced compared with themodel group; Qing-Fei-Pei-Yuan granules group compared with the AZT group andTang Herb group,0.5＜2-△△ct＜2;It explains that JNK2mRNA expression of this threegroups has no difference.8.①When Qing-Fei-Pei-Yuan granules group and AZT group compared with themodel group,2-△△ct＜0.5，It explains that ERK1mRNA expression of this two groupswas significantly reduced compared with the model group; Qing-Fei-Pei-Yuangranules group compared with the Tang Herb group,0.5＜2-△△ct＜2，It explains thatERK1mRNA expression of this two groups has no difference;Qing-Fei-Pei-Yuan granules group was significantly increased compared with the AZT group,2-△△ct＞2, Itexplains that ERK1mRNA expression of Qing-Fei-Pei-Yuan granules group wassignificantly increased compared with the AZT group.②When Qing-Fei-Pei-Yuangranules group and AZT group compared with the model group,2-△△ct＜0.5，Itexplains that ERK2mRNA expression of this two groups was significantly reducedcompared with the model group; the Tang Herb group compared with the model group,0.5＜2-△△ct＜2, It explains that ERK2mRNA expression of this two groups has nodifference; Qing-Fei-Pei-Yuan granules group compared with the AZT group andTang Herb group,2-△△ct＜0.5， It explains that ERK2mRNA expression ofQing-Fei-Pei-Yuan granules group was significantly reduced compared with the AZTgroup and Tang Herb.Conclusion:1. Qing-Fei-Pei-Yuan granules and each treatment group can improve the generalstation of mouse models. It explains that Qing-Fei-Pei-Yuan granules have a goodtherapeutic effect for immunodeficiency and pulmonary infection.2. The p38、JNK and ERK protein and mRNA expression of the model mice’ lungtissue is significant increase compared with the normal group,It reveals that theyparticipate the occurrence of immunodeficiency and pulmonary infection.3. Qing-Fei-Pei-Yuan granules improve immune function and controlinflammation process by inhibiting the transcription factor targets of p38、JNK andERK.4. MAPK signaling pathway may be one of the molecular mechanisms thattreatment of immunodeficiency and pulmonary infection for Qingfeipeiyuan particles.