Research On The Mechanism Of Qinbaiqingfei Concentrated Pills In The Treatment Of Mycoplasma Pneumonia Based On Intestinal Flora And Mucosal Immunity

Objective: Through the establishment of a Mycoplasma pneumoniae pneumonia(MPP)model,based on the intestinal flora-mucosal immunity theory,the use of related technologies to explore the regulation of Qinbaiqingfei Concentrated Pills on the mucosal immune function and intestinal microecology of MPP rats.Method : First: Take the blank rats as the normal reference and azithromycin(Xisumei)as the control drug.After administering the MPP rats,the MP amplification vector amount in the lungs,HE pathological sections of lung tissue,serum IL-10,TNF-α,IL-4,INF-γ,etc.The level of inflammatory factors and the ratio of Th1/Th2 of immune cytokines were tested to evaluate the efficacy of Qinbaiqingfei Concentrate Pills.Subsequently: Using 16 Sr DNA technology and the Illumina Nova sequencing platform to sequence the intestinal flora of rats,through the complexity analysis of the intestinal flora of rat feces and comparison of multiple samples,the differences and dominant bacteria of the samples at different classification levels between the groups were analyzed.The phylogenetic tree analysis of species type and genus level identifies species with significant differences.To clarify the ability of Qinbai to adjust the structure of intestinal flora in MPP rats.Further: Use GC-MS technology to study the content of short-chain fatty acids in rat feces,determine the content of acetate,propionate,butyrate,isobutyric acid,and isovaleric acid in rat feces,and explore Qinbai’s MPP rats The ability to regulate and control short-chain fatty acids.Final: ELISA technology and Western blotting method were used to detect serum and organ tissues of rats in this experiment.Serum LPS,TGFβ1,intestinal mucosa SIgA were detected by ELISA kit,lung tissue MyD88,IKKα,IκBα,NFκB p65 and colon tissue HDAC1,HDAC3,and NFκB p65 were tested for protein expression to clarify the regulatory pathways of Qinbaiqingfei Concentrated Pills on humoral and cellular immunity in vivo.Results:(1)Qinbai Qingfei Concentrate Pills can inhibit the proliferation of MP in the body,and significantly reduce the content of pathogens in the lung tissue;at the same time,Qinbai can repair lung tissue damage caused by MPP,reduce the thickness of alveolar walls and restore alveolar shape,Inhibit the accumulation of eosinophils,maintain the normal morphology of connective tissue and blood vessels in the lungs,etc.;restore the high expression of serum IL-10,TNF-α,IL-4,INF-γ and other inflammatory factors in rats caused by MPP,and increase the Th1/Th2 ratio.(2)From the perspective of MPP,the abundance of Firmicutes,General Archaea and Actinomycota at the phylum level can decrease and the Bacteroides phyla can be increased,and Qinbai has a significant callback effect on this.At the class level,the decline of Salmonella,Bacillus,Propionibacterium,Klosterella,Actinomycetes and the increase of Bacteroides,the regulation of Qinbazai class level was significantly better than that of the azithromycin group.Bacteroides,Clostridium streptococcus,Campylobacter,Erysipelas,Clostridia＿UCG-014,Bifidobacterium,Acholeplasmatales,Rhododactylidae,etc.are higher than the blank group at the order level,Bacteroides,RF39,Acetobacterales,Coryneformes,Staphylococcales and other abundance decreased,Qinbai and azithromycin can produce significant protection,mainly concentrated in Erysipelas,Clostridia＿UCG-014,Bifidobacterium,Coryneforme four bacteria,and Qinbai pairs have nothing to do There is less intervention of bacteria,and it basically does not interfere with the abundance of other bacteria while regulating the unbalanced bacteria.The family level results are similar to the former.The relative abundance of Muribaculaceae,Prevotellaceae,Enterococcus,Aerococcaceae,Acetobacteraceae,Corynebacteriaceae,Staphylococcaceae,etc.in the blank group Decrease;the relative abundances of the rest of the Veronococcus,Bifidobacteria,Cholesteromycetes,and Defluviitaleaceae in the blank group are higher than those in the model group.Combining the species evolution tree to study the genus level,Qinbai not only has the ability to call back unbalanced bacteria,but also has a strong ability to promote the growth of lactobacilli.(3)GC-MS analysis confirmed the ability of Qinbai to significantly restore the acetate,propionate,butyrate,and isobutyric acid content of SCFAs.(4)Qinbai Qingfei Concentrated Pills can significantly inhibit the expression of MyD88,IKKα,IκBα,NFκB p65 in lung tissues,inhibit serum LPS,TGFβ1,and promote the secretion of SIgA in the intestinal mucosa,confirming that Qinbai has an effect on cellular immunity.Inhibition and promotion of humoral immunity.The experiment also confirmed that Qinbai can inhibit the expression of HDAC1,HDAC3,NFκB p65 and other proteins in colon tissue.Conclusion:(1)Qinbai can remove Mycoplasma pneumoniae in rat lung tissue and repair damaged lung tissue.(2)Qinbai can inhibit the secretion of IL-10,TNF-α,IL-4,INF-γ and other cytokines,and increase the ratio of Th1/Th2.(3)Qinbai can adjust the dysregulated colony of MPP rats at various classification levels,increase the abundance of gram-positive bacteria,promote the production of lactobacilli,and reduce the secretion of gram-negative bacteria LPS;The secretion of metabolites acetate,propionate,butyrate,etc.plays a role in promoting.(4)Qinbai can promote the secretion of SIgA through the regulation of intestinal flora and the regulation of TGFβpathway.(5)Qinbai can inhibit HDAC1 and HDAC3 by regulating SCFAs,thereby inhibiting the expression of NF-κB pathway proteins MyD88,IKKα,IκBα,and NFκB p65 in the lung and intestine mucosa.