基于生物信息学和实验探究艾灸对肺纤维化的作用及其机制  

Effect of moxibustion on pulmonary fibrosis based on bioinformatics and experiments and the mechanism

作　　者：董怀洋 叶雨萌 李包娟 张䶮之 DONG Huai-yang;YE Yu-meng;LI Bao-juan;ZHANG Yan-zhi(Department of Pharmacology,School of Pharmacy,Xinjiang Medical University,Urumqi 830011)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011

出　　处：《中南药学》2022年第6期1217-1225,共9页Central South Pharmacy

基　　金：国家自然科学基金项目(No.81760767);新疆维吾尔自治区自然科学基金面上项目(No.2017D01C204);新疆医科大学博士启动基金(No.2019-017);新疆自治区重点实验室经费(No.XJDX1713)。

摘　　要：目的基于现代生物信息技术,预测艾灸干预肺纤维化的作用靶点和通路,并通过动物实验验证作用效果及其机制。方法通过生物信息预测得到艾灸干预肺纤维化的作用靶点,蛋白互作获得作用的关键靶点,GO和KEGG将通路可视化,采用AutoDock4.2.6对接软件进行验证。博来霉素4 U·kg^(-1)构建肺纤维化模型,连续给药干预28 d。每日观察小鼠外观,测量小鼠饮食、体质量,计算肺指数,HE和Masson染色观察肺组织病变及纤维化程度,ELISA测定相关因子,蛋白免疫印迹法检测相关蛋白表达,荧光定量法测定mRNA水平。结果得到关键靶点8个,关键通路为EGFR/JAK/STAT1,分子对接显示4种核心成分有极好的对接活性。动物实验结果表明艾灸可改善小鼠体质量,提高存活率,HE和Masson染色结果显示艾灸后炎症细胞和胶原细胞减少。艾灸组小鼠的肺系数、肺组织Hyp和血清IL-6、CCL2炎症因子水平显著降低,肺组织EGFR、JAK、STAT1、p-STAT1蛋白表达显著降低,相关mRNA水平显著降低。结论艾灸可通过抑制EGFR/JAK/STAT1通路延缓肺纤维化进程。Objective To predict the targets and pathways of moxibustion for pulmonary fibrosis based on modern bioinformatics,and to clarify the effect and mechanism by animal experiments.Methods The targets of moxibustion for pulmonary fibrosis were predicted by bioinformatics.The pathways were visualized by GO and KEGG and validated by molecular docking with AutoDock4.2.6.The pulmonary fibrosis model was established with 4 U·kg^(-1) bleomycin and administered continuously for 28 days.The mice were observed every day,and their diet and body weight were measured.HE and Masson staining were used to observe the degree of lung tissue lesions and fibrosis.ELISA was used to detect related inflammatory factors.Western blot was used to detect the expression of related proteins,and fluorescence quantitative detection for mRNA levels.Results Eight key targets were identified,and the key pathways was EGFR/JAK/STAT1.Molecular docking showed the 4 key components had excellent docking activity.Animal experiments showed that moxibustion improved the body weight and survival rate of mice.HE and Masson staining showed that the inflammatory cells and collagen cells in the moxibustion group were reduced.The lung coefficient,Hyp in the lung tissue,and serum IL-6 and CCL2 inflammatory factor levels in the moxibustion group were significantly reduced.Western blot showed down-regulation of the protein expressions of EGFR,JAK,STAT1,and p-STAT1.The related mRNA levels were significantly decreased.Conclusion Moxibustion can delay the process of pulmonary fibrosis by inhibiting EGFR/JAK/STAT1 pathways.

关 键 词：肺纤维化模型 艾灸 生物信息分析 动物实验验证 EGFR/JAK/STAT1通路 

分 类 号：R285[医药卫生—中药学]