The Role Of Transcription Factor NF-κb, AP-1in Qingfeipeiyuan Particles Intervention Immunodeficiency And Pulmonary Infection BALB/C Mice Model’ Immune And Inflammatory Processes

Objective:By studying effects of the Qingfeipeiyuan particles on the expression of NF-κB/AP-1in immunocompromised and pulmonary infected mice, to explore the possible molecularmechanisms. in order to provide experimental evidence for the Applications ofQingfeipeiyuan particles, also do some exploration for the development of effective drugsfor curing immunodeficiency and pulmonary infection.Methods:1. Animal groupingFirst, divide the60female BALB/c mice into two groups, the normal group has12mice,the buildding group which is to be built the module has48mice,when modulesucessfulled made, divided the buildding group randomLy into four groups according to therandom number table: model group, AZT group, Tangcao tablet group, Qingfeipeiyuanparticles group,In order to distinguish we dyed them with Picric Acid solution.2.Animal model makingThe normal group was given0.9%saline according0.25mL/10g standards byIntraperitoneal injection after three days, the buildding group vaccinate an equal volume ofFLV virus.According to FLV virus pathogenic characteristics, determine the modeling periodwas21d.When immunocompromised mouse model is successfull, injected0.03-0.05mL10%chloral hydrate into abdominal,then gave them50μl the Ⅲ type pneumococcalbacterial suspension using intranasal method, According to pathogenic characteristics of theⅢ type Streptococcus pneumoniae, determine the modeling period is7days,It is time totreat them by drenching.3. AdministrationAfter one day from successful modeling,then begin to fed mice of each group withcorresponding drug, the AZT group was given AZT suspension（0.08g/kg.d）for14days,Tangcao tablet group was given Tangcao tablet suspension（2.56g/kg.d）for14days,Qingfeipeiyuan particles group was given Qingfeipeiyuan particles（2g/kg.d）for14days,the normal group and model group were given0.9%saline for14days, The frequency ofdrenching of all groups are one time per day,and dose is0.2mL10g-1.4. Index detectionObserve the general condition of the mice such as activity、eating、hair color and measurebody weight 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 partof these lung tissue were measured the NF-κB/AP-1protein expression by Western Blotmethod, the other part were measured the NF-κB/AP-1mRNA expression by Real time PCRmethod.Results:1.The general condition of mice wered improved in different degrees in each treatmentgroup Comparison with the model group compared with the model group.2. NF-κB p65、 AP-1c-fos、 c-jun mRNA and protein expression of the lung tissue aresignificantly higher than the model group(P <0.05).3. NF-κB p65protein expression of the lung tissue of Qingfeipeiyuan particles groupand other groups are lower than the modle group,and the difference was statisticallysignificant (P <0.05);It is also lower than AZT group and TangHerb group, the difference arestatistically significant (P <0.05).4. When Qingfeipeiyuan particles group and AZT group compared with the modelgroup,2-△△ct＜0.5，It explains that NF-κB p65mRNA expression of Qingfeipeiyuan particlesgroup and AZT group was significantly reduced compared with the model group; TangHerbgroup compared with the model group,0.5＜2-△△ct＜2，It explains that NF-κB p65mRNAexpression of these two groups has no difference; Qingfeipeiyuan particles group comparedwith TangHerb group,2-△△ct＜0.5，It explains that NF-κB p65mRNA expression ofQingfeipeiyuan particles group was significantly reduced compared with the TangHerbgroup; Qingfeipeiyuan particles group compared with AZT group,0.5＜2-△△ct＜2，It explainsthat NF-κB p65mRNA expression of these two groups has no difference.5.AP-1c-fos protein expression of the lung tissue of Qingfeipeiyuan particles groupand other groups are lower than the modle group,and the difference was statisticallysignificant (P <0.05);It is higher than AZT group,but lower than TangHerb group, thedifference are statistically significant (P <0.05).6. When Qingfeipeiyuan particles group and other groups compared with the modelgroup,2-△△ct＜0.5，It explains that AP-1c-fos mRNA expression of Qingfeipeiyuan particles group and AZT group was significantly reduced compared with the model group; TangHerbgroup compared with the Qingfeipeiyuan particles group and the model group,0.5＜2-△△ct＜2，It explains that AP-1c-fos mRNA expression of this three groups has no difference;Qingfeipeiyuan particles group compared with the AZT group,2＜2-△△ct，It explains thatAP-1c-fos mRNA expression of Qingfeipeiyuan particles group was significantly increasedcompared with the AZT group.7. AP-1c-jun protein expression of the lung tissue in Qingfeipeiyuan particles groupand AZT group are lower than the modle group,and the difference was statisticallysignificant (P <0.05);but it has no difference between the modle group and TangHerbgroup(P＞0.05); AP-1c-jun protein expression of the lung tissue of Qingfeipeiyuan particlesgroup is lower than the AZT group and TangHerb group, and the difference was statisticallysignificant (P <0.05).8. When Qingfeipeiyuan particles group、TangHerb group and AZT group comparedwith the model group,2-△△ct＜0.5，It explains that AP-1c-jun mRNA expression of this threegroups was significantly reduced compared with the model group; Qingfeipeiyuan particlesgroup compared with the TangHerb group0.5＜2-△△ct＜2，It has no difference between thetwo groups. When compared with AZT group,2-△△ct＜0.5,It explains that AP-1c-jun mRNAexpression of Qingfeipeiyuan particles group was significantly reduced compared with theTangHerb group and AZT group.Conclusion:1. Qingfeipeiyuan particles can improve the general station of mouse models,Thisverify the effectiveness of Qingfeipeiyuan particles in therapying immunocompromised andpulmonary infected.2.The activation of transcription factor NF-κB and AP-1are related to theimmunodeficiency and pulmonary infection.3. Qingfeipeiyuan particles can efficiently inhibit the activation of transcription factorNF-κB and AP-1, regulate related immune and inflammatory mediators, thereby controlinflammation, improve the symptoms of immunocompromised and pulmonary infected. Itillustrates that the mechanism that Qingfeipeiyuan particles treat immunocompromised andpulmonary infected micemay be associated with the transcription factor NF-κB and AP-1.4. Transcription factor NF-κB, AP-1may be potential targets for the treatment ofmmunodeficiency and pulmonary infection.