Preliminary Study On The Treatment And Mechanisms Of Bulleyaconitine A On Anti-inflammatory Of Lung In Allergic Asthma Mice

Objective:In this article,anti-inflammatory effects on allergic asthma mice were observed by preparing a mouse model of allergic asthma and treating with Bulleyaconitine A.Furthermore,to observe the related mechanisms and anti-inflammatory effect of Bulleyaconitine A on LPS-induced RAW264.7 cells.Methods: 60 female mice were divided arbitrarily into 6 experimental groups(n =10): Control group;OVA group;OVA + Bulleyaconitine A(0.16 mg/kg);OVA +Bulleyaconitine A(0.32 mg/kg);OVA + Bulleyaconitine A(0.48 mg/kg);dexamethasone(1 mg/kg).In all groups except the control one,mice were exposed to sensitization on days 0,7 and 14 by intraperitoneal(i.p.)injection of 25 ?g OVA emulsified with 2 mg Al(OH)3 in 200 ?l phosphate buffered saline(PBS)(pH 7.4).On days 22 to 28,mice were challenged with inhalation of 5% OVA solution in saline,for30 min by Bulleyaconitine A.DEX was injected i.p.for 7 days starting from day 22 till the end of experiment Bulleyaconitine A was given orally for 7 days starting from day22 till the end day.Mice were sacrificed 24 h after the last challenge.(1)After sacrifice,we performed enzyme-linked immunosorbent assay(ELISA)to analysis serum immunoglobulin just like IgE and IgG.(2)The cytokine(IL-4,IL-10,TNF-? and MCP-1)concentrations were determined by ELISA.(3)Besides this,inflammatory cell counts and classification in bronchoalveolar lavage fluid(BALF)were determined.(4)Hematein and Eosin staining were performed to show inflammatory cell infiltration and mucus production.(5)Furthermore,Lungs were used for preparation of homogenates.(6)We treated RAW264.7 with Bulleyaconitine A,then performed cell viability assay to evaluate cytotoxicity.(7)We use ELISA to evaluate cytokine(IL-1?,IL-6,TNF-? and MCP-1)expression in cell supernatant.(8)Western blot were used to explore effectsand mechanisms of Bulleyaconitine A on NF-?B activation.Results: In conclusion,our results proposed that in asthmatic mice,Bulleyaconitine A effectively reduced ovalbumin-induced IgE and IgG in surum.Treatment with Bulleyaconitine A decreased production of cytokines(IL-4,TNF-? and MCP-1),and inflammatory cell counts(eosinophils,lymphocytes and macrophages)in BALF.Histological studies demonstrated that Bulleyaconitine A substantially inhibited OVA-induced eosinophilia in lung tissue and airway tissue.Western blotting studies demonstrated that Bulleyaconitine A substantially inhibited NF-?B1 and PKC-? protein level.Conclusion: 1.Bulleyaconitine A can alleviates the lung inflammation in mice with allergic asthma,it relieve Airway inflammatory response via inhibit PKC-?/NF-?B Signaling pathway.2.Bulleyaconitine A attenuated the release inflammatory cytokines in the IL-1?,IL-6,TNF-? and MCP-1 in RAW264.7 cells induced by LPS.3.The anti-inflammatory effects of Bulleyaconitine A was through blocking LPS-mediated NF-?B signaling pathways in RAW264.7 cells.