补元汤干预COPD大鼠Th17细胞分化及其相关炎症因子分泌的影响

The Effect of Buyuan Decoction on the Differentiation of Th17 Cells and the Secretion of Related Inflammatory Factors in COPD Rats

目的 观察补元汤对COPD模型大鼠T细胞辅助细胞17(Th17)及相关细胞因子之间的影响。方法 将36只SD大鼠随机分为对照组,模型组,补元汤低、中、高剂量组,氨茶碱组,每组6只。使用烟熏联合气道内滴注脂多糖(LPS)建立COPD大鼠模型,造模4周后开始药物干预,连续给药4周。分别于末次烟熏及末次给药后检测大鼠肺功能:TV、PEF、f、RT。HE染色观察大鼠肺组织病理改变;实时荧光定量PCR(qPCR)和Western blot检测大鼠肺组织RORγt、IL-17A、IL-21的mRNA及蛋白表达;免疫荧光双标记法检测肺组织RORγt/IL-17A、STAT3/IL-17A及IL-21/IL-17A共定位表达情况,流式细胞术检测肺组织中CD3^(+) CD4^(+) IL17A^(+) Th17细胞占CD4^(+) T细胞比例。结果 与模型组比较,补元汤各剂量组与氨茶碱组大鼠TV、PEF、f、RT指标均明显改善,肺组织病理损伤显著减轻,RORγt、IL-17A、IL-21的mRNA及蛋白含量明显下降,RORγt/IL-17A、STAT3/IL-17A及IL-21/IL-17A共定位表达量明显降低,CD3^(+) CD4^(+) IL17A^(+) Th17细胞占CD4^(+) T细胞比例显著减少。结论 补元汤可能是通过调控Th17细胞分化,抑制炎症细胞因子的分泌,从而延缓COPD的免疫炎症进展。

Objective To explore the effect of Buyuan Decoction on T cell helper cell 17(Th17)and related cytokines in COPD model rats.Methods 36 SD rats were randomly divided into control group,model group,low,medium,and high dose groups of Buyuan decoction,and aminophylline group,with 6 rats in each group.The COPD rat model was established using smoking com⁃bined with intratracheal instillation of lipopolysaccharide(LPS).After 4 weeks of modeling,drug intervention began and the drug was administered continuously for 4 weeks.Detect the pulmonary function of after the last smoking and administration:TV,PEF,f,RT.HE staining was used to observe the pathological changes of rat lung tissue;Real time fluorescence quantitative PCR(qPCR)and Western blot was used to detect the rat mRNA and protein expression of RORγt,IL-17A and IL-21;Immuno⁃fluorescence double labeling method for detecting lung tissue Co localization expression of RORγt/IL-17A,STAT3/IL-17A,and IL-21/IL-17A;Flow cytometry was used to detect the proportion of CD3^(+)CD4^(+)IL-17A^(+)Th17 cells in lung tissue.Re⁃sults Compared with the model group,the TV,PEF,f,and RT indicators of rats in each dose group of Buyuan Decpction and the aminophylline group were significantly improved,and the pathological damage to lung tissue was significantly reduced.The mRNA and protein content of RORγt,IL-17A and IL-21 significantly decreased,and the co-localization expression levels of RORγt/IL-17A,STAT3/IL-17A,and IL-21/IL-17A were significantly reduced,and the proportion of CD3^(+)CD4^(+)IL-17A^(+)Th17cells was significantly reduced.Conclusion Buyuan decoction may delay the progression of immune inflammation in COPD by regulating Th17 cell differentiation and inhibiting the secretion of inflammatory cytokines.