Effect Of Electroacupuncture Modulation Of Cholinergic Anti-inflammatory Pathway On Type 2 Intrinsic Lymphocytes And Their Inflammatory Factors Of IL-5 And IL-13 In Asthmatic Mice

Objectives:By observing the effects of electroacupuncture(EA)on anti-inflammation and inhibition of allergic reactions in asthma model mice,as well as the contents of ILC2s and related inflammatory factors interleukin(IL)-5 and IL-13 in lung tissues and blocking CAP,the mechanism of action of EA in the treatment of asthma was explored to provide a scientific basis for clinical acupuncture treatment of asthma.Methods:(1)Animal grouping and model preparation:40 female Balb/c mice were randomly divided into four groups:normal control group,model group,model+EA group and model+α-BGT(α7nAChR receptor antagonist)+EA group,10 mice in each group.The asthma model was established by using a mixture of ovalbumin(OVA)and aluminum hydroxide dry powder for intraperitoneal injection sensitization combined with nebulized inhalation stimulation.(2)EA treatment:The mice in the model+EA group and the model+α-BGT+EA group were given EA to the"Dazhui","Fengmen"and"Feishu"on both sides,respectively,using sparse and dense waves.The frequency was 2Hz/15Hz,intensity 0.5m A,once a day for 30min,for 7 days.(3)Indexes:asthma symptom scores were observed;hematoxylin-eosin staining(HE),periodate-Scherff staining(PAS)and Masson staining were used to observe lung histopathology;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of immunoglobulin E(IgE),OVA-sIgE,inflammatory factors IL-5 and IL-13 in serum and lung tissues;flow cytometry was used to detect the levels of inflammatory factors IL-5 and IL-13 in serum and lung tissues.levels;flow cytometry to detect ILC2s and IL-5~+ILC2s,IL-13~+ILC2s.Results:(1)Model evaluation:compared with normal control group,asthma symptoms were obvious in the model group,and the score was significantly increased(P<0.01);bronchial lumen narrowing,massive inflammatory cell infiltration in airways,increased mucus secretion,inflammation score and Muc/Pbm,Wcol/Pbm were significantly increased(P<0.01);IgE,OVA-sIgE content were significantly increased(P<0.01).(2)EA effect:compared with the model group,the asthma symptoms were reduced and the scores were significantly lower in the model+EA group(P<0.01);the bronchial lumen narrowing and airway inflammatory cell infiltration were improved,and the inflammation scores,Muc/Pbm and Wcol/Pbm were decreased(P<0.01);the expression contents of IgE and OVA-sIgE were significantly down-regulated(P<0.01);the serum and IL-5 and IL-13 in lung tissue were decreased(P<0.01);the distribution ratio of ILC2s,IL-5~+ILC2s and IL-13~+ILC2s in lung tissue was decreased(P<0.01).(3)Blocking CAP:compared with the model+EA group,mice in the model+α-BGT+EA group had aggravated asthma,increased scores(P<0.05),increased airway inflammatory cell infiltration;significantly higher levels of IgE and OVA-sIgE(P<0.01);significantly higher levels of IL-5 and IL-13 in serum and lung tissues(P<0.01);lung tissue ILC2s,IL-5~+ILC2s,and IL-13~+ILC2s ratios were elevated(P<0.01).Conclusion:EA can improve airway inflammation,reduce allergic reaction,and relieve asthma symptoms.Its anti-inflammatory and inhibitory effect on allergic reaction may be related to the CAP-mediated downregulation of the amount of ILC2s in lung tissue.