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探讨阻断IL-17A对小鼠肺损伤后肺纤维化发生的预防作用及其机制。博来霉素造成小鼠肺损伤7天后给予抗IL-17A中和性抗体治疗,第14天取材。利用HE和天狼星红染色法观察肺组织炎症及胶原沉积情况;利用生化检测试剂盒检测肺组织中羟脯氨酸及肺泡灌洗液中胶原含量;利用Kaplan-Meier法统计小鼠生存率;利用ELISA方法检测肺泡灌洗液中IL-17A、TGF-β1、IL-13、IFN-γ表达水平;利用免疫印迹分析方法检测肺组织中p65NF-κB、p50NF-κB、COX-2、5-LOX、15-LOX的表达或活化。结果显示:与模型对照组比较,阻断IL-17A能够降低胶原沉积、减少羟脯氨酸及胶原含量、提高小鼠生存率,并能抑制炎症反应,降低IL-17A、TGF-β1、IL-13,升高IFN-γ、COX-2、5-LOX、15-LOX表达水平,抑制p65NF-κB活化但促进p50NF-κB活化。结果提示,阻断IL-17A能预防肺损伤后肺纤维化发生,其机制与促进p50NF-κB活化及其下游分子表达、抑制急性炎症转变为慢性炎症反应有关。
To investigate the preventive effect and mechanism of blocking IL-17A on pulmonary fibrosis in mice after lung injury. Bleomycin induced lung injury in mice 7 days after anti-IL-17A neutralizing antibody treatment, the first 14 days of accession. The lung tissue inflammation and collagen deposition were observed by HE staining and Sirius red staining. The contents of hydroxyproline and alveolar lavage fluid in lung tissue were detected by biochemical detection kit. The survival rate of mice was calculated by Kaplan-Meier method. The levels of IL-17A, TGF-β1, IL-13 and IFN-γ in BALF were detected by ELISA. The expressions of p65NF-κB, p50NF-κB and COX-2,5-LOX in lung tissue were detected by Western blot , 15-LOX expression or activation. The results showed that blocking IL-17A could reduce the collagen deposition, decrease the content of hydroxyproline and collagen, increase the survival rate of mice and inhibit the inflammatory reaction and IL-17A, TGF-β1, IL -13, increase the expression of IFN-γ, COX-2,5-LOX and 15-LOX, inhibit the activation of p65NF-κB and promote the activation of p50NF-κB. The results suggest that blocking IL-17A can prevent pulmonary fibrosis after lung injury, and its mechanism is related to the promotion of p50NF-κB activation and its downstream molecule expression, inhibition of acute inflammation into chronic inflammation.