摘要
目的基于特异性转录因子T-bet/GATA3通路探讨温肺止咳颗粒治疗哮喘的疗效及可能机制。方法将60只雄性SD大鼠,分为空白组、哮喘模型组、中药组、地塞米松组,每组各15只。通过HE染色镜下观察各组大鼠肺组织的病理改变;测定各组血清白细胞介素-4(Interleukin-4,IL-4)和干扰素-γ(Interferon-γ,INF-γ)水平;Westem-Blot检测各组肺组织T细胞特异性转录因子T-bet(T-box expressed in T cells,T-bet)和GATA3(GATA-binding protein-3,GATA3)水平;RT-PCR检测各组肺组织T-bet mRNA、GATA3 mRNA、INF-γmRNA、IL-4 mRNA水平。结果空白组大鼠肺组织肺泡上皮细胞排列整齐,无坏死和脱落,肺泡壁无水肿或充血,肺泡腔及支气管腔内无渗出物,间质未见炎细胞浸润;哮喘模型组肺泡间质中见中度的炎细胞浸润(++),肺泡壁充血(+);中药组及地塞米松组肺泡间质及支气管黏膜下可见轻度炎细胞浸润(+)。哮喘模型组大鼠血清中IL-4含量较空白组明显增高,INF-γ含量明显降低,差异有统计学意义(P<0.01);中药组及地塞米松组大鼠血清IL-4的含量较哮喘模型组明显降低,INF-γ明显增高,差异有统计学意义(P<0.01)。哮喘模型组较空白组肺组织中T-bet蛋白表达水平更低,GATA3蛋白表达水平更高,T-bet/GATA3比值低,差异有统计学意义(P<0.01);中药组及地塞米松组较哮喘模型组肺组织中T-bet蛋白表达增高,GATA3蛋白表达水平降低,T-bet/GATA3比值增高,差异有统计学意义(P<0.01)。哮喘模型组大鼠GATA3 mRNA和IL-4 mRNA表达较空白组增高,T-bet mRNA和INF-γmRNA表达降低,差异有统计学意义(P<0.05);中药组和地塞米松组大鼠较哮喘模型组GATA3 mRNA和IL-4 mRNA表达降低,INF-γmRNA和T-bet mRNA表达明显增高,差异有统计学意义(P<0.05)。结论对哮喘大鼠应用温肺止咳颗粒治疗,可有效改善大鼠肺泡间质及支气管黏膜炎症反应,通过调控T-bet/GATA3通路,可改善大鼠血清中IL-4和INF-γ,起到改善哮喘大鼠症状作用。
Objective To explore the possible mechanism of Wenfei Zhike Granules in the treatment of asthma based on the T-bet/GATA3 pathway.Methods Sixty male SD rats were included and divided into a blank group,an asthma model group,a Chinese medicine group,and a dexamethasone group,with 15 rats in each group.The pathological changes in the lung tissue of 4 groups were observed by hematoxylin-eosin(HE)staining.The levels of serum interleukin-4(IL-4)and interferon-γ(INF-γ)in each group were determined.The expression levels of T-bet and GATA-3 in the lung tissue of rats in each group was detected by Western blot,and the mRNA expression levels of the T-bet,GATA3,INF-γ,and IL-4 in the lung tissue were detected by real-time polymerase chain reaction(RT-PCR).Results In the lung tissue of rats in the blank group,there was no obvious degeneration,necrosis,and shedding of alveolar epithelial cells,no congestion or edema in the alveolar wall,no exudate in the alveolar cavity and bronchial cavity,and no inflammatory cell infiltration in the interstitium.In the asthma model group,moderate inflammatory cell infiltration(++)and alveolar wall congestion(+)were observed in the alveolar interstitium.In the Chinese medicine group and the dexamethasone group,mild inflammatory cell infiltration(+)was observed in the alveolar interstitium and bronchial mucosa.As compared with the blank group,the content of serum IL-4 in the asthma model group was increased,and that of INF-γwas decreased(P<0.01).As compared with the asthma model group,the content of serum IL-4 in the Chinese medicine group and the dexamethasone group was decreased,and that of INF-γwas increased(P<0.01).As compared with the blank group,the protein expression level of T-bet in the lung tissue of rats in the asthma model group was decreased,that of GATA3 was increased,and the ratio of T-bet/GATA3 was decreased(P<0.01).As compared with the asthma model group,the protein expression level of T-bet in the lung tissue of rats in the Chinese medicine group and the dexamethasone group was increased,that of GATA3 was decreased,and the ratio of T-bet/GATA3 was increased(P<0.01).As compared with the blank group,the mRNA expression levels of GATA3 and IL-4 in the asthma model group were increased,whereas those of T-bet and INF-γwere decreased(P<0.05).Conclusion The application of Wenfei Zhike Granules in rats with asthma can effectively improve the inflammatory response of alveolar interstitium and bronchial mucosa.By regulating the T-bet/GATA3 pathway,it can improve the serum IL-4 and INF-γ,thus promoting the syndromes of rats with asthma.
作者
侯海慧
唐艳芬
苏成程
洪玲玲
章晶晶
HOU Hai-hui;TANG Yan-fen;SU Cheng-cheng;HONG Ling-ling;ZHANG Jing-jing(Pneumology Department,Nantong Hospital of Traditional Chinese Medicine,Nantong Jiangsu 226000)
出处
《世界中西医结合杂志》
2022年第12期2416-2421,2439,共7页
World Journal of Integrated Traditional and Western Medicine
基金
2019年南通市卫健委科研项目(QA2019017)
2020年南通市科技局科技计划项目(JCZ20091)。
