| Aim:We established asthma models with RSV infection and OVA as allergen, to observe the effects of Gubenfangxiao decoction (GBFXD) on asthmatic mice and the immunity way.Method:RSV was first prepared,and the virulence is100TCID50.54female BALB/c mouses were divided randomly into six groups,including the control group,the asthma group, the montelukast group,the low dose of GBFXD group,the GBFXD in medium dose,and the high dose of GBFXD.In addition to the normal control group,the mice were given RSV intranasal infection.The mice were sensitized with intraperitoneal OVA injection on week1and week3ofter infection, and with atomization inhaled OVA3weeks later.Atomization was made once a day and for30minutes once a time in10days.The control group was stimulate asthma with normal saline,to observe whether these is restless,fecal incontinence,rapid breathing and other symptoms.The control group and asthma group were lavaged by normal saline,others were administered intragastrically with GBFXD or montelukast after atomization each day.All mice were sacrificed24hours after the last administration to gather the BALF for figuring out cell counts and cell classifications, and to observe the pathological changes of lungs.We use ELISA to setting-out the levels of IL-4and IFN-γ.Examining the expression of STAT1and STAT3through western blot.Results:(1)Asthma group compare with control group:the quantity of cells and eosinophils is apparently higher (p<0.01), the counts of eosinophils and neutrophils are a little higher, IL-4levels were higher (P<0.01) and IFN-y were lower (P<0.01);(2)Asthma group is higher in the expression of p-STAT1and p-STAT3in lungs than the control group;(3)Administered with GBFXD or montelukast,the counts of leukocyte are lower in BALF (P<0.05),Lymphocytes and basophils were obviously lower (P<0.01),which demonstrated that GBFXD takes a part in the process of airway inflammatory reaction,and is beneficial for asthma;(4)Contrast to asthma group,GBFXD and montelukast groups have a lower IL-4levels and a higher IFN-y levels (P<0.05),but there is little difference between montelukast group and the GBFXD groups.It is showed that GBFXD can obviously increase IFN-y/IL-4to regulate the function of Thl and Th2;(5)The mice in control group has normal lung and bronchial structures.There is also no inflammatory exudation around,without mucosal edema and hyperemia, wall and smooth muscle structure is also normal.In asthma group,bronchial mucosa have heavy hyperemia and edema, inflammatory cells infiltration,wall and smooth muscle have a hyperplasia and hypertrophy.Administered with GBFXD or montelukast,inflammatory infiltration is significantly improved,the thickness of the wall and smooth muscle is reduced,it is indicated that GBFXD can change the pathology of asthma mice in airway.(6)Signal transduction and transcription activating factor (STAT) plays an important role in the inflammation and immunity regulation of asthma. GBFXD low dose group have a poor efficacy, but in other groups the expression p-STAT1and p-STAT3are lowered (P<0.05), which proved that GBFXD inhibits the immunity of asthma by restraining the phosphorylation of STAT1and STAT3in the signaling pathway.Conclusion:(1) GBFXD of preventing asthma attack might be ascribed to the decreasing of airway inflammation.(2) GBFXD also can decrease IL-4levels and increase IFN-y levels, what is good for Th1/Th2imbalance.(3) Activation of STAT1(p-STAT1) and STAT3(p-STAT3) involve in the mechanisms of asthma, GBFXD can reduce the expression of p-STAT1and p-STAT3, relieve inflammation and do good for the treatment of asthma. |
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