Objective:To explore the presence of inflammatory memory in the lung tissue of mice with cough variant asthma(CVA)and the mechanism of Wuhu decoction in preventing and treating CVA.Methods:Among the 90 SPF-grade femal...Objective:To explore the presence of inflammatory memory in the lung tissue of mice with cough variant asthma(CVA)and the mechanism of Wuhu decoction in preventing and treating CVA.Methods:Among the 90 SPF-grade female BALB/c mice,20 were randomly selected as the blank group,while the remaining 70 were sensitized by intraperitoneal injection of ovalbumin(OVA)combined with aluminum hydroxide and then exposed to OVA by nebulization to establish the CVA mouse model.From the conctrol group and the model group,10 mice were randomly selected to detect airway responsiveness using an animal ventilator.Successfully modeled mice were then randomly divided into 7 groups:a CVA model group,a CVA re-challenge group,low-,medium-,and high-dose Wuhu decoction groups,and a positive control(dexamethasone)group,with 10 mice in each group.After 30 days of rest,except for the conctrol group and the model group,mice in the other groups underwent a second OVA nebulization challenge and received corresponding drug interventions once a day for 10 consecutive days before further analysis.Pathological changes in lung tissue were observed using periodic acid-schiff(PAS)and Masson staining methods.Serum levels of tumor necrosis factor-α(TNF-α),interleukin-4(IL-4),interferon-γ(IFN-γ),and interleukin-10(IL-10)were detected using enzyme-linked immunosorbent assay(ELISA).The expression of T-bet mRNA and GATA-3 mRNA in lung tissue was determined by real-time quantitative polymerase chain reaction(Real-time PCR).Results:The results of airway responsiveness indicated successful establishment of the model.Compared with the control group,the model group exhibited severalkey pathological changes including inflammatory cellinfiltration around the lung tissue,epithelialmetaplasia,airway collagen deposition,increased airway mucus accumulation index and airway collagen deposition index(P<0.05),elevated serum TNF-αand IL-4 expression(P<0.05),decreased IFN-γand IL-10 levels(P<0.05),decreased T-bet mRNA levels(P<0.05),increased GATA-3 mRNA levels(P<0.05),and a reduced T-bet mRNA/GATA-3 mRNA ratio(P<0.05)in lung tissue.When compared to the CVA re-challenge group,the Wuhu decoction groups and the dexamethasone group effectively alleviated pathological changes in lung tissue,reduced the airway mucus accumulation index and airway collagen deposition index(P<0.05),decreased serum TNF-αand IL-4 expression(P<0.05),increased serum IFN-γand IL-10 levels(P<0.05),upregulated T-bet mRNA levels(P<0.05),downregulated GATA-3 mRNA levels(P<0.05),and increased the T-bet mRNA/GATA-3 mRNA ratio(P<0.05)in lung tissue.Conclusion:The lung tissue of mice with cough variant asthma exhibits inflammatory memory,and Wuhu decoction may intervene in this inflammatory memory by restoring the balance between T-bet and GATA-3,thereby correcting the imbalance in Th1/Th2 immunity and exerting a preventive and therapeutic effect on cough variant asthma.展开更多
文摘Objective:To explore the presence of inflammatory memory in the lung tissue of mice with cough variant asthma(CVA)and the mechanism of Wuhu decoction in preventing and treating CVA.Methods:Among the 90 SPF-grade female BALB/c mice,20 were randomly selected as the blank group,while the remaining 70 were sensitized by intraperitoneal injection of ovalbumin(OVA)combined with aluminum hydroxide and then exposed to OVA by nebulization to establish the CVA mouse model.From the conctrol group and the model group,10 mice were randomly selected to detect airway responsiveness using an animal ventilator.Successfully modeled mice were then randomly divided into 7 groups:a CVA model group,a CVA re-challenge group,low-,medium-,and high-dose Wuhu decoction groups,and a positive control(dexamethasone)group,with 10 mice in each group.After 30 days of rest,except for the conctrol group and the model group,mice in the other groups underwent a second OVA nebulization challenge and received corresponding drug interventions once a day for 10 consecutive days before further analysis.Pathological changes in lung tissue were observed using periodic acid-schiff(PAS)and Masson staining methods.Serum levels of tumor necrosis factor-α(TNF-α),interleukin-4(IL-4),interferon-γ(IFN-γ),and interleukin-10(IL-10)were detected using enzyme-linked immunosorbent assay(ELISA).The expression of T-bet mRNA and GATA-3 mRNA in lung tissue was determined by real-time quantitative polymerase chain reaction(Real-time PCR).Results:The results of airway responsiveness indicated successful establishment of the model.Compared with the control group,the model group exhibited severalkey pathological changes including inflammatory cellinfiltration around the lung tissue,epithelialmetaplasia,airway collagen deposition,increased airway mucus accumulation index and airway collagen deposition index(P<0.05),elevated serum TNF-αand IL-4 expression(P<0.05),decreased IFN-γand IL-10 levels(P<0.05),decreased T-bet mRNA levels(P<0.05),increased GATA-3 mRNA levels(P<0.05),and a reduced T-bet mRNA/GATA-3 mRNA ratio(P<0.05)in lung tissue.When compared to the CVA re-challenge group,the Wuhu decoction groups and the dexamethasone group effectively alleviated pathological changes in lung tissue,reduced the airway mucus accumulation index and airway collagen deposition index(P<0.05),decreased serum TNF-αand IL-4 expression(P<0.05),increased serum IFN-γand IL-10 levels(P<0.05),upregulated T-bet mRNA levels(P<0.05),downregulated GATA-3 mRNA levels(P<0.05),and increased the T-bet mRNA/GATA-3 mRNA ratio(P<0.05)in lung tissue.Conclusion:The lung tissue of mice with cough variant asthma exhibits inflammatory memory,and Wuhu decoction may intervene in this inflammatory memory by restoring the balance between T-bet and GATA-3,thereby correcting the imbalance in Th1/Th2 immunity and exerting a preventive and therapeutic effect on cough variant asthma.
