Mechanism Of Jianpi Buifei Huatan Prescription In Treatment Of Brochial Asthma Based On “intestinal Flora-SCFAs/histamine-MCs”

Objective:Based on the correlation of intestinal flora to disease outcome and its impact on immunity to respiratory diseases,this study explored the mechanism of Jianpi Bufei Huatan prescription for the treatment of clinical remission of asthma(CRA)by intestinal flora.Methods:According to the principle of complete randomness,the 48 female BLAB/c mice were divided into 6 groups,namely blank control group(HSE group),model group(MOD group),traditional Chinese medicine with model group(TCM group),western medicine with model group(MST group)),co-feeding model group(C-MOD group),and co-feeding traditional Chinese medicine with model group(C-TCM group),with 8 rats in each group.These groups who participated in the CRA model were sensitized with OVA+AL(OH)3 and nebulized with OVA.Invasive pulmonary ventilation function test was used to detect FEV1/FVC and FEF 25＿75/FVC under anesthesia to evaluate pulmonary ventilation function;Hematoxylin and eosin staining was used to detect the pathological changes of lung and colon tissues;Sandwich-ELISA was used to detect IL-33 and histamine.The protein expression of leukotriene C4(LTC4)and mast cell growth factor receptor(C-KIT)were detected by Western blotting(WB).The expression of LTC4 m RNA was detected by real-time fluorescence quantitative assay(PCR).The content of intestinal flora was detected by 16 S r DNA sequencing technology.short-chain fatty acid(SCFAs)were detect by gas chromatography-mass spectrometry(GC-MS).Immunofluorescence(IF)was used to label mast cell chymotrypsin 1(MAPT-1).Result:1.Pulmonary function and pathology: Compared with HSE group,MOD group had decreased pulmonary ventilation function(P<0.05)and a lot of pathological changes in lung and colon(P<0.05);Compared with the MOD group,the lung ventilation function of TCM and MST groups were improved(P<0.05),and the pathological conditions of lung and colon were relieved;2.IL-33、LTC4:The levels of IL-33,LTC-4 protein and m RNA in MOD group were higher than those in HSE group(P<0.05).The levels of IL-33,LTC-4 protein and m RNA in TCM and MST groups were lower than those in MOD group(P<0.05).3.Intestinal flora,SCFAs and histamine in feces and histamine in serum: Compared with HSE group,the abundance of Bacteroides and Actinomycetes decreased(P<0.05),the abundance of Firmicutes increased(P<0.05),the F/B ratio increased(P<0.05),the total amount of SCFAs decreased(P<0.05),the acetic acid and propionic acid decreased significantly(P<0.05),and the histamine levels in serum and feces increased(P<0.05)in MOD group.Compared with MOD group,TCM group had higher abundance of Bacteroides and Actinomycetes(P<0.05),lower abundance of Firmicutes(P<0.05),lower F/B ratio(P<0.05),the level of SCFAs in feces changed,and lower histamine levels in serum and feces(P<0.05).4.Pathology,intestinal flora and their metabolites in co-feeding experiments: Compared with the MOD group,the pathological changes of lung and intestine were relieved,the histamine level in serum was decreased(P<0.05),and the histamine level in feces tended to decrease in C-MOD group(P<0.05).Compared with TCM group,C-TCM group had more serious pathological changes in lung and intestine(P<0.05)),higher histamine levels in feces and serum(P<0.05).Compared with C-MOD group,C-TCM group showed no difference in flora richness compared with SCFAs(P>0.05),decreased histamine levels in serum and decreased histamine levels in feces(P<0.05).5.Mast cell markers(MAPT-1,C-KIT): Compared with HSE group,the expression of MAPT-1 and C-KIT in MOD group was higher(P<0.05).Compared with MOD group,the expression of MAPT-1 and C-KIT in C-MOD group,TCM group and C-TCM group were lower(P<0.05).Compared with TCM group,the expression of MAPT-1 and C-KIT were higher in C-TCM group(P<0.05).Conclusion:1.Jianpi Bufei Huatan Recipe can effectively alleviate the inflammatory cell infiltration in lung and colon tissues of ARS mice,and inhibit airway remodeling;2.Jianpi Bufei Huatan Recipe can decrease the level of IL-33 in serum and the expression of LTC4 protein and m RNA in airway of ARS mice,and improve the pulmonary ventilation function.3.Jianpi Bufei Huatan Recipe can regulate the abundance of intestinal flora,histamine and SCFAs in ARS mice.4.The therapeutic effect of Jianpi Bufei Huatan Recipe on ARS mice through "intestinal flora-SCFAs/histamine-MCs" axis.