Study On The Effect And Mechanism Of Huatan Huoxue Formula Intervention On Asthma By Inhibiting ILC2s

Objective:In this paper,intraperitoneal injection and inhalation of ovalbumin(Ovalbumin,OVA)were used to replicate the mouse model of asthma,and the effect of Huatan Huoxue Formula on the intervention of asthma through ILC2 s and its mechanism in the migration pathway of ILC2 s in the lung-gut axis were investigated.Provide a basis for research and development in asthma.Method:1 Study on the effect of Huatan Huoxue Formula and its disassembled prescriptions on airway mucus hypersecretion in asthmatic mice In this study,64 BALB/c female mice were randomly divided into 8 groups: normal group,model group,dexamethasone group,Huatan Huoxue Formula group,Sanziyangqintang group,Taohongsiwutang group,Sanziyang Qintang + Taoren Honghua group,Sanziyangqintang + Siwutang group,8 in each group.On the 1st,8th,and 15 th days,intraperitoneal injection of 2m L OVA sensitizing fluid was performed,and on the 21 st and27th days,inhalation of 2% OVA challenge fluid was used to replicate the asthma mouse model;each group was given a corresponding drug by intragastric administration 1h before the atomization;Observe the behavior of mice and the incubation period of asthma;on the28 th day,take the serum,BALF and lung tissue of each group of mice.The rapid Wright-Jimsa staining method was used to observe the changes in the total number of white blood cells and the number of various types of white blood cells in BALF;the HE staining method was used to observe the pathological changes of lung tissue;the PAS staining method was used to observe the proliferation of lung goblet cells;The content of total Ig E and OVA-Ig E in serum and the content of IL-4,IL-9,IL-13 and MUC5 AC in BALF were detected by ELISA kit.2 Effect of Huatan Huoxue Formula on the migration of ILC2 s in the lung-gut axis of asthmatic mice The establishment and administration of asthma mouse model are the same as Part 1.On the 28 th day of the experiment,the BALF,lung tissue,small intestine tissue and mesenteric lymph nodes of each group of mice were taken;the content of IL-25,IL-33,TSLP,IL-6 and AREG in BALF was detected by ELISA kit;Quantitative reverse transcription PCR(RT-q PCR)detection of mi R-155,mi R-146 a,IL-25,Sphk1 in lung tissue and IL-25,Sphk1 m RNA expression in small intestine tissue;flow cytometry was used to detect ILC2 s,lung and mesenteric lymph nodes in lung tissue.The content of n ILC2 s and i ILC2 s in the medium;the expression of S1PR1 and ILC2s(GATA3 +EPCAM+IL-17RB)in the small intestine tissue was observed by immunofluorescence.Result:1 Study on the effect of Huatan Huoxue Formula and its disassembled prescriptions on airway mucus hypersecretion in asthmatic mice Both Huatan Huoxue Formula and its decomposed formulas can effectively improve asthma-like behavior induced by OVA in asthmatic mice,increase the incubation period of asthma(P<0.01),reduce the level of inflammatory cell infiltration in lung tissue,reduce the total number of white blood cells in BALF and the number of various types of leukocytes(P <0.05,P <0.01)and reduce the levels of total Ig E and OVA-Ig E in serum(P<0.01);meanwhile,Huatan Huoxue Formula and its decomposed formulas can alleviate the proliferation of goblet cells in the lung tissue of asthmatic mice,and can significantly reduce the content of inflammatory factors IL-4,IL-9 and IL-13 in BALF that are associated with high mucus secretion(P <0.05,P <0.01),and can significantly reduce the level of MUC5 AC,the main component of mucin(P <0.01),thereby inhibiting the high secretion of airway mucus in asthmatic mice.In addition,these results show that the effect of Huatan Huoxue Formula is better than that of each group.2 Effect of Huatan Huoxue Formula on the migration of ILC2 s in the lung-gut axis of asthmatic mice In the asthma model of OVA induced by Huatan Huoxue Formula,the content of IL-25,IL-33,TSLP,IL-6 and AREG in BALF of asthmatic mice can be significantly reduced(P<0.05,P <0.01).And down-regulated mi R-155 and increased mi R-146 a m RNA expression in lung tissue(P <0.01);flow cytometry detection of lung tissue ILC2 s showed that the proportion of ILC2 s decreased significantly(P <0.01).At the same time,through the detection of lung-intestinal axis ILC2 s migration-related indicators,it was found that Huatan Huoxue Formula can reduce the expression of IL-25 m RNA and Sphk1 m RNA in lung and small intestine tissue(P <0.01).Reducing the expression of S1PR1 and ILC2s(GATA3 + EPCAM + IL-17RB)in small intestine tissues.Through flow cytometry detection of i ILC2 and n ILC2 s in lung and small intestine tissues,it was found that Huatan Huoxue Formula can reduce the content of i ILC2 s in lung and small intestine tissues(P<0.01),and can reduce the content of n ILC2 s in the small intestine tissue(P <0.01),but has no obvious effect on the generation of n ILC2 s in the lung tissue.Conclusion:Huatan Huoxue Fang and its disassembled prescriptions can improve airway inflammation by inhibiting mucus hypersecretion in asthmatic mice,the mechanism of its intervention in asthma may be to block the migration of i ILC2 s in the small intestine to the lungs and reduce the content of ILC2 s in lung tissue,Thereby the type 2 immune response is reduced.