芒柄花素磺酸钠通过MAPKS信号通路缓解OVA诱导哮喘小鼠肺部炎症

Study on the effect and mechanism of sodium formononetin-3’-sulfonate on OVA-induced asthma mice

关键词:  芒柄花素磺酸钠  哮喘  卵清蛋白  小鼠

Key words:  sodium formononetin-3’-sulfonate  asthma  OVA  mice

摘要:

[目的] 观察芒柄花素磺酸钠对卵清蛋白（OVA）诱导哮喘小鼠的保护作用及其机制。[方法] 购买SPF级C57BL/6小鼠60只，并随机分6组，每组10只，具体组别如下：空白对照组（CON组）、模型组（OVA组）、芒柄花素磺酸钠低剂量（MBHS-L组）、芒柄花素磺酸钠中剂量（MBHS-M组）、芒柄花素磺酸钠高剂量（MBHS-H组）和地塞米松组（DEX组），使用OVA诱导建立小鼠哮喘模型。实验主要评价小鼠哮喘症状并进行评分、计量支气管肺泡灌洗液（BALF）中炎症细胞数量（Diff-Quick染色法）、镜下观察肺组织的病理学变化[苏木精-伊红（HE）染色法]、分析BALF中超氧化物歧化酶（SOD）活性和肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）、白细胞介素6（IL-6）和丙二醛（MDA）水平（ELISA法和化学发光法）、分析肺组织p-ERK1/2，p-JNK和p-p38MAPK蛋白表达水平（免疫荧光法和Western blot法）。[结果] 与OVA组比较，哮喘症状评分、BALF炎症细胞数量、TNF-α、IL-1β和MDA水平以及p-ERK1/2，p-JNK和p-p38MAPK蛋白表达水平在MBHS组和DEX组均明显降低（P＜0.05），而SOD的活性明显升高（P＜0.05），肺组织病理学变化明显改善。[结论] 芒柄花素磺酸钠可能通过MAPKS信号通路缓解OVA诱导哮喘小鼠的肺部炎症。

Abstract:

[Objective] To observe the protective effect of sodium formononetin-3’-sulfonate on OVA-induced asthma mice and its mechanism. [Methods] Sixty SPF C57BL/6 mice were purchased and randomly divided into 6 groups with 10 mice in each group. The specific groups are as follows：control group（CON group），model group（OVA group），low dose of sodium formononetin-3’-sulfonate（MBHS-L group），medium dose of sodium formononetin-3’-sulfonate（MBHS-M group），high dose of sodium formononetin-3’-sulfonate（MBHS-H group） and dexamethasone group（DEX group）. A mouse asthma model was established using OVA. The experiment was mainly to evaluate the asthma symptoms of mice and score them，measure the number of inflammatory cells in bronchial alveolar lavage fluid（BALF）（Diff-Quick staining），observe the pathological changes of lung tissue under microscope（HE staining），and analyze the activity of SOD and the levels of TNF-α，IL-1β，IL-6 and MDA in BALF（ELISA method），protein expression levels of P-ERK1/2，p-JNK and p-p38MAPK in lung tissue were analyzed by immunofluorescence and Western blot. [Results] Compared with OVA group，asthma symptom score，BALF inflammatory cell number，TNF-α，IL-1β and MDA levels，p-ERK1/2，p-JNK and p-p38MAPK protein expression levels were significantly decreased in MBHS group and DEX group（P<0.05），while SOD activity was significantly increased（P<0.05）. The changes of lung histopathology were significantly improved. [Conclusion] Sodium formononetin-3’-sulfonate may relieve lung inflammation in mice with OVA-induced asthma through MAPKS signaling pathway.