Exploring The Intervention Mechanism Of Wuhu Decoction On RSV-induced Asthma DCs Autophagy Based On STAT3 Signaling Pathwa

Objective: To investigate the mechanism of autophagy of DCs induced by RSV through STAT3 signaling pathway in the devel-opment of asthma,and to elucidate the mechanism of autophagy of DCs induced by Wuhu decoction through regulating STAT3 signaling pathway to improve airway inflammation and airway remodeling in mice with virus-induced asthma,reduce airway hyp-erresponsiveness and inhibit the development of "phlegm",so as to provide a new theoretical and experimental basis for the pre-vention and treatment of this disease in Chinese medic ine.Methods:(1)RSV combined with OVA inhalation was used to st-imulate as thma.After succes sful modeli ng,the mice were rand omly divid ed into normal group,model gro up,STAT3 inhib itor grou p(STATTI C),and STAT3 in du ce r gr oup(col i ve l in),wit h 10 mice in each grou p,and corresponding drug intervention for 2we eks.The ai rway react i vi t y va lues of mi ce were de tec te d by an an imal vent il at or;HE,PAS and Mas s on st aini ng methods were use d to ob s erv e the pa thol og ic al ch anges of mous e lung tis s ue,g-oblet cells secreted mucus and airway collagen deposition;ELISA method was used to detect serum IL-6,IL-10 and IL-17 content;Real-ti me PC R met hod wa s us ed to det erm ine the exp res s ion le-vel s of TGF-β 1 m R N A,α-S M A m R N A and STAT-3 m R N A in lu-ng tissue;transmission electron microscopy was used to detect lung tissue autophagosomes;The levels of ATG5 and SQSTM1 and the expression of STAT3 and p-STAT3 in lung tissue DCs we re observed by Western Blot and immunocytochemical staining,res pe ct i ve ly.(2)The RS V co m poun d OVA as t hm a mou s e mode l was co nstru ct ed.Aft er succes s ful modeli ng,the mice were rand-oml y di vide d int o norm al group,mo del group,pos it ive drug co-ntrol group(dexamethas one),STAT3 inhibitor group(STATTIC),and STAT3 induc er group(coli v elin)and Wuhu decocti on low-d-ose,med i um-dos e and hi gh-d os e gro ups,10 in each group,an d were given correspond ing drug in terventi on for 2 week s.The air way reactivity val ues of mice were det ected by an animal venti-lator;HE,PAS and Masson staini ng met hods were used to obse-rve the pathological changes of mou se lung tis sue,gobl et cel ls secreted mucus and airway collagen deposition;ELISA method was us ed to det ect seru m IL-6,IL-10 and IL-17 cont ent;R eal-t i-me PC R me t ho d was us ed to det e rm i ne the expre s s i on le vel s of TGF-β 1 m RNA,α-SMA m RNA and STAT-3 m RNA in lung ti sse;Trans mis si on el ectron micros copy was us ed to detect lung tis su e autophag osomes;The level s of ATG5 and SQSTM 1 and the expres sion of STAT3 and p-STAT3 in lung tiss ue DCs were obs erved by Western Blot and immunohistochemistry,respectively.Results :(1)The experiment al results showed that airway reacti-vity increased in the model group,lung immunohistochemistry showed narrowing of bronchial lumen,mas sive inflammatory cell infil tratio n around blood vess els and bronchi,excess ive hyperpl-asia of cupular epithelial cells and extensive deposition of air way col l age n,increas ed seru m lev el s of IL-6 and IL-17 and de-creas ed level s of IL-10 compare d wit h the no rm al gro up,TGF-β 1m RNA and α-SMA m RNA expression levels were significantly increas e;The STAT3 inhibi tor group(S TATTIC)reduc ed airway hyperrespo nsiv enes s,reduced lung ti ssue damage,reduced serum I L-6 and IL-17 le vel s,in cre as e d IL-10 le ve ls,and red u ce d TG F-β1 m RNA and α-SMA m RNA expression levels in lung tissues compa red wit h th e mod el gr o up,whi l e the STAT3 induce r gr o u p(colivelin)showed no stat istical significance com pared with the model group;The result s of transmis sion electron mi cros copy and autophag y protein ass ay showed that the aut oph ag y level of DCs in lun g ti s s ues of mi ce in th e model gr oup inc reas e d,as shown by the increase in the number of autoph agi c vesi cles,th e increase in the level of DCs autophagy protein ATG5 and the decr-ease in the level of SQSTM1;compared with the model group,the autophagy level of STAT3 inhibitor group(STATTIC)increa sed further,wh il e the aut ophagy level of STAT3 inducer group(collivel in)decreas ed,and the res u lt s of imm unocytochemi cal were bas i call y cons is t ent wi t h the abo ve res ul ts;The ex press i on level s of p-STAT3/STAT3 in DCs of mouse lung tissues in the model group were significantly increased.Compared with the model group,the expres s ion level s of p-STAT3/S TAT3 in the STAT3 inhibi tor gro up(S TATTIC)we re si gni fi cant l y de cre as ed,and the expression levels of p-STAT3/STAT3 in the STAT3 indu cer group(colivelin)were significantly increased.The results of PCR method and immunocytochemical staining were basically consistent with the above results.(2)The experimental results showed that after the intervention of Wuhu decoction,it could reduce the airway reactivity of asthmatic mice;improve the lung tissue damage and inflammatory cell infiltration in mice,reduce the expression of IL-6 and IL-17 in serum,increase the expression of asthma protective cytokine IL-10,and improve airway inflammation;effectively reduce airway mucus secretion and collagen deposition under airway epithelial cells in asthmatic mice,reduce lung It also reduced the expression levels of TGF-β1 m RNA and α-SMA m RNA in lung tissues and improved airway remodeling;increased the autophagy level of DCs in lung tissues,increased the level of ATG5 protein and decreased the level of SQSTM1 protein;down-regulated the expression levels of p-STAT3/STAT3 in DCs in lung tissues and inhibited the STAT3 pathway.Conclusion:(1)In RSV-induced asthma,the level of autophagy in DCs rises and the STAT3 signaling pathway was activated,causing asthma airway hyperresponsiveness and airway inflammation to occur,leading to airway remodeling and thus affecting asthma development.(2)Wuhu Decoction further upregulate the autophagy level of DCs by inhibiting the STAT3 signaling pathway,reduce the airway hyperresponsiveness of asthma improves airway inflammation and airway remodeling,reduce the occurrence and development of“phlegm ”,and plays a role in the prevention and treatment of asthma.