基于TWEAK/Fn14信号通路探讨大黄酚对OVA诱导哮喘小鼠的保护作用

Study on the protective effect of rhubarphol on ovalbumin induced asthmatic mice based on TWEAK/Fn14 signal pathway

关键词:  大黄酚  卵清蛋白  哮喘  TWEAK/Fn14信号通路  小鼠

Key words:  rhubarphol  ovalbumin  asthma  TWEAK/Fn14 signal pathway  mouse

摘要:

[目的] 观察大黄酚对卵清蛋白（OVA）诱导哮喘小鼠模型的保护作用，并探讨肿瘤坏死因子样凋亡弱诱导因子（TWEAK）/成纤维细胞生长诱导因子14（Fn14）信号通路与其保护作用的关系。[方法] 60只C57BL/6小鼠按体质量随机分为正常对照组（CON组）、模型组（OVA组）、地塞米松阳性对照组（DEX组）、大黄酚低剂量组（RHU-L组）、大黄酚中剂量组（RHU-M组）和大黄酚高剂量组（RHU-H组），每组10只。实验结束后，对各组小鼠的哮喘症状进行评分；Diff-Quick染色法计量各组小鼠支气管肺泡灌洗液（BALF）中炎症细胞数。苏木精-伊红（HE）染色法观察肺组织病理学变化；酶联免疫吸附法检测各组小鼠BALF中白细胞介素-4（IL-4）、白细胞介素-5（IL-5）、肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的含量；免疫组化法检测各组小鼠肺组织中TWEAK蛋白的表达水平。蛋白免疫印迹法检测各组小鼠肺组织TWEAK/Fn14信号通路蛋白TWEAK和Fn14的表达水平。[结果] 大黄酚可明显降低哮喘症状评分（P<0.05），减少哮喘小鼠BALF中总细胞、嗜酸性粒细胞、中性粒细胞和淋巴细胞的数量（P<0.05），改善肺组织的病理学变化，降低BALF中IL-4、IL-5、TNF-α和IL-1β炎症因子的水平（P<0.05），减少肺组织中TWEAK和Fn14蛋白表达（P<0.05）。[结论] 大黄酚对OVA致哮喘小鼠模型具有较好的保护作用，其可能与调节TWEAK/Fn14信号通路有关。

Abstract:

[Objective] To observe the protective effect of rhubarphol on asthmatic mice induced by ovalbumin (OVA), and explore whether the mechanism of its effect is related to TWEAK/Fn14 signal pathway.[Methods] Sixty C57BL/6 mice were randomly divided into the normal control group (CON group), the model group (OVA group), the dexamethasone positive control group (DEX group), the low dose rhubarphol group (RHU-L group), the medium dose rhubarphol group (RHU-M group) and the high dose rhubarphol group (RHU-H group) according to their body weight (10 mice/group). At the end of the experiment, asthma symptoms of each group were scored. Diff-quick staining method was used to measure the number of inflammatory cells in BALF of mice in each group. Histopathological changes of lung were observed by HE staining. The contents of IL-4, IL-5, TNF-α and IL-1β in BALF were detected by ELISA. Immunohistochemical method was used to detect the level of TWEAK in lung tissues of mice in each group. The expression levels of TWEAK and Fn14 were by Western blot in lung tissue.[Results] Rhubarphol could significantly reduce asthma symptom score (P<0.05), and decreased the total number of cells, eosinophils, neutrophils and lymphocytes in BALF of asthmatic mice (P<0.05). Rhubarphol improved pathological changes in lung tissue, and reduced the levels of IL-4, IL-5, TNF-α and IL-1β in BALF (P<0.05). Rhubarphol decreased the expression of TWEAK and Fn14 in lung tissues (P<0.05).[Conclusion] Rhubarphol has protective effect on OVA induced asthma mice, it may be related to regulation of TWEAK/Fn14 signal pathway.