| Objective:Chronic bronchitis is one of common diseases in breathe system.This is a unpeculiarly inflammation of bronchia catarrh and tissue around.It mainly comport as coughing,spitting or with gasping cross-outbreak.With the population aging and environment polluted, Chronic bronchitis's outbreak is manifolding,which is seriously harmful to the health of people.The Lung-QI deficiency syndrome (LQDS) is the major syndrome type of Chronic bronchitis.Recently,many scholars who with morden technology method,studied extensively on LQDS.They made some progresses in this field.They both think that CB are induced by the unbalance of proinflammatory factor and antinflammatory factor in the view of inflammation.The source of proinflammatory factor and antinflammatory factor can divide to three type:T cell secretion,mononuclear secretion,neutrophil secretion.TH1 and TH2 were differentiated by T cell,which could secret proinflammatory factor and antinflammatory factor. TH1 and TH2 were correlated with CB closely.But the study about the infection of TH1 and TH2's related cytokines is not reported.The disease model was induced by reforming smokig method and copied chronic bronchitis with LQDS. By examing the content of IL-4 and IFN-γin lung blood and tissue,this study explored the treatment role and mechanism of traditional Chinese medication and will reveal further the essence of chronic bronchitis with LQDS,which is hoped to present the evidence for the prevention and treatment of this disease in clinic.Methods: 50 healthy SD rats (Paroecious about half) raised in the experimental environment about one week were randomly divided in to five groups (10 per-group):①control group;②CB model group;③Bufeitang prednisone group;④Buqihuxuefang low-dose group;⑤Buqihuoxuefang high-dose group.Based on literature,the disease model was induced by reforming smoking method and copied chronic bronchitis with LODS.The model rats were placed in a smoke room specially made for 1m×1m×1m.The paring,sawdust and tobacco leaf each fifty gram were burn and fumed the smoke,which was bid,30 minutes every time,and kept on 30 day.The rats of normal group were fed in non-smoke condition.From the second day after instillation,five groups were continuously filled with saline or medicine for 21 days.The normal group and model group were filled with NS(10ml/kg),low-dose group and high-dose group were drenched with Chinese crude drug elixation differently( 12.8g/kg,25.6/kg,7 or 14 times the dosage of the adult's clinical drug consumption);prednisone group were drenched with prednsion(8.36g/kg,7 times the dosage of the adult's clinical drug consumption).Preparation of sample:All rats in five groups were treated once a day for 21 days.The total rats in each group were anesthesiaed and sampled blood from abdomen main artery,centrifuged 5min with 3500r/min,and stored them with the temperature of -20℃for examination.Lung tissue about 1cm×1cm×1cm size taken from the middle of lung,was fixed with 4% formaldehyde,embedded with paraffin commonly,and made slice.Preparation of homogenate of lung tissue:washed the lung tissue with iced saline ,absorbed the fluid of surface,weighed the wet weight is 0.3g,and then dripped precooling saline,milling-in with the machine of homogenate ,ad got 10% homogenate of lung tissue.Cetrifuged 5 min with 6000r/min with 4℃,extracted the supernatant,and stored them with the temperature -20℃for examination.By HE and SP,observe the change of lung tissue's pathomorph. By the radioactive immunochemistry,examing the content of IL-4 and IFN-γin lung blood and tissue.Results:1 Observation of the common conditionNormal group ate and drunk normally,reacted sensitivively,the fur was luster,the urine and stool were normal,with no death.Model group scrunched,coughed and breathed difficulty,had listlessness and withered fur.High-dose group and Low-dose group gradually had the better mind,fur and appetite,breathed smoothly,and their activity increased;prednisone group alsohad better condition,but inferior to High-dose group and Low-dose group.2 Observation of morphological lung tissue2.1 Observation with eyesThe lung's appearance of normal group was common with color pink,surface smooth,elasticity good and lobes of lung edge sharpness.The lung's appearance of model group was abnormal with color gray-black,surface concavity,without burnish and ecchymosis were seen.The lung tissues of every group to treat were abnormal with color redness and eccymosis sometimes,but they all had obvious changes to compare with the model group. 2.2 Observation of HEThe result of phthomorphology manifested that in normal group mucous membrane of bronchiole epithelia was integrity,cells lined up orderly,bronchiole dissepiment and alveoli had no distinctive pathomorphologic alteration under light microscope.In model group,bronchioles cavity was enlarged,vessel wall was thicked significantly,part of epithelial cellular necrosis and abscission,there was a lot of chronic inflammatory cells infiltration and venule with stagnant blood.Every group which was medicated all had obvious changes,especially the high-dose group.2.3 Observation of Immunohistochemical methodThe CB rats in normal group IL-4 were better positive expression in the Mucous membrane of bronchiole epithelium hyalomitome(Fig.11);The CB rats in model group IL-4 were weaker positive expression in the Mucous membrane of bronchiole epithlium hyalomitome(Fig.12);Positive signal were all butty.The CB rats in low-dose group and bufeitang group IL-4 were weaker positive expression in the Mucous membrane of bronchiole epithelium hyalomitome(Fig.13,14);The CB rats in High-dose group IL-4 were better positive expression in the Mucous membrane of bronchiole epithelium hyalomitome.Be similar with nomal group,positive reaction mainly expressed in Mucous membrane of bronchiole epithelium hyalomitome(Fig.15).Semiquantitative result showed:The IL-4 contents in lung tissue of CB rats in model group were significantly lower than the nomal group(P<0.01);The IL-4 contents in lung tissue in three therapeutic group was higher than the model group(P<0.01 or P<0.05 );Compared three therapeutic groups,The IL-4 contents in lung tissue in high-does group and low-dose group were higher than bufeitang group (P<0.01);Compared with high-dose group and low-dose group,there were no statistics meanin(P>0.05).The CB rats in normal group IFN-γwere weaker positive expression in the Mucous membrane of bronchiole epithelium hyalomitome(Fig.16);The CB rats in model group IFN-γwere better positive expression in the Mucous membrane of bronchiole epithlium hyalomitome(Fig.17);Positive signal were all butty.The CB rats in low-dose group and bufeitang group IFN-γwere better positive expression in the Mucous membrane of bronchiole epithelium hyalomitome(Fig.18,19);The CB rats in High-dose group IFN-γwere weaker positive expression in the Mucous membrane of bronchiole epithelium hyalomitome.Be similar with nomal group,positive reaction mainly expressed in Mucous membrane of bronchiole epithelium hyalomitome(Fig.20), Semiquantitative result showed:The IFN-γcontents in lung tissue of CB rats in model group were significantly higher than the nomal group(P<0,01);The IFN-γcontents in lung tissue in three therapeutic group was lower than the model group(P<0.01 or P<0.05 );Compared three therapeutic groups,The IFN-γcontents in lung tissue in high-does group were higher than low-dose group (P<0.01);Compared with low-dose group and bufeitang group,there were no statistics meanin(P>0.05).3 The IL-4,IFN-γcontents and the rate of TH1/TH2 change in serum and lung tissue of each group rats3.1 The IL-4 contents change in serum and lung tissue of each group ratsThe result showed: the IL-4 contents in blood serum of CB rats in model group were significantly lower than the nomal group(P<0.01);The IL-4 contents in blood serum in three therapeutic groups were higher than the model group(P<0.01);Compared three therapeutic groups,The IL-4 contents in blood serum in high-dose group were higher than bufeitang group and low-dose group(P <0.01 or P <0.05);there were no statistics meaning in low-dose group and bufeitang group(P>0.05).The IL-4 contents in lung tissue of CB rats in model group were significantly lower than the nomal group(P<0.01);The IL-4 contents in lung tissue in three therapeutic groups were higher than the model group(P<0.01);Compared three therapeutic groups,The IL-4 contents in high-dose group and low-dose group were higher than bufeitang group(P <0.01 or P <0.05);There were no statistics meaning in high-dose group and low-dose group(P>0.05).Table23.2 The IFN-γcontents change in serum ang lung tissue of each group ratsThe result showed: the IFN-γcontents in blood serum of CB rats in model group were significantly higher than the nomal group(P<0.01);The IFN-γcontents in blood serum in three therapeutic groups were lower than the model group(P<0.01);Compared three therapeutic groups, the IFN-γcontents in blood serum in high-dose group and low-dose group were lower than bufeitang group (P<0.01 or P<0.05);the IFN-γcontents in blood serum in high-dose group were lower than low-dose group(P<0.01).The IFN-γcontents in lung tissue of CB rats in model group were significantly higher than the nomal group(P<0.01);the IFN-γcontents in lung tissue in three therapeutic groups were lower than the model group(P<0.01);Compared three therapeutic groups, the IFN-γcontents in lung tissue in high-dose group and low-dose group were lower than bufeitang group( P<0.01);the IFN-γcontents in lung tisssue in high-dose group and low-dose group were were no statistics meaning(P>0.05). Table33.3The rate of TH1/TH2 change in serum and lung tissue of each group ratsThe result showed: the rate of TH1/TH2 in blood serum of CB rats in model group were significantly higher than the nomal group( P<0.01);the rate of TH1/TH2 in blood serum in three therapeutic groups were lower than model group(P<0.01);compared three therapeutic groups, the rate of TH1/TH2 in blood serum in high-dose group and low-dose group were lower than bufeitang group(P<0.01); the IFN-γcontents in blood serum in high-dose group and low-dose group were no statistics meaning(P>0.05).The rate of TH1/TH2 in lung tissue of CB rats in model group were significantly higher than the nomal group(P<0.01);the rate of TH1/TH2 in lung tissue in three therapeutic groups were lower than model group(P<0.01 or P<0.05);compared three therapeutic groups, the rate of TH1/TH2 in lung tissue in high-dose group were lower than low-dose group and bufeitang group(P<0.01);the IFN-γcontents in lung tissue in low-dose group and bufeitang group were no statistics meaning(P>0.05).Conclusion:1 Under the condition of CB Lung-QI Deficiency Syndrome,the resistance inflammation cytokine IL-4 content in blood serum and lung tissue are decreased,the promotion inflammation cytokine IFN-γare increased.They co-induced unpeculiarly inflammation reaction of CB Lung-QI Deficiency Syndrome.This alleosis probably causes the pathogenesis of CB Lung-QI Deficiency Syndrome.2 Buqihuoxue recipe brings up the IL-4 contents in blood serum and lung tissue,adjust the IFN-γcontents and the rate of TH1/TH2 in blood serum and lung tissue,thus attenuates the promation of CB Lung-QI Deficiency Syndrome's unpeculiarly inflammation reaction by cytokine network, gain the ends that preventing and curing CB Lung-QI Deficiency Syndrome.
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