基于蛋白组学探讨麦冬皂苷D改善肺纤维化的作用机制  

Study on the mechanism of ophiopogonin D in improving pulmonary fibrosis based on proteomics

作　　者：陈婷 包晟川 李京涛 魏海梁 惠毅[1] 闫曙光[1,2] CHEN Ting;BAO Sheng-chuan;LI Jing-tao;WEI Hai-liang;HUI Yi;YAN Shu-guang(Teaching and Research Department of Gastroenterology,School of Basic Medicine,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Shaanxi Provincial Key Laboratory of Gastrointestinal Disease and Syndrome Formulae,Xianyang 712046,China;First Department of Hepatology,The Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712046,China;First Departments of General Surgery,The Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]陕西中医药大学基础医学院胃肠病教研室,咸阳712046 [2]陕西省胃肠病证方药重点研究室,咸阳712046 [3]陕西中医药大学附属医院肝病一科,咸阳712046 [4]陕西中医药大学附属医院普外一科,咸阳712046

出　　处：《中国新药杂志》2024年第4期372-382,共11页Chinese Journal of New Drugs

基　　金：国家自然科学基金资助项目(81703974);陕西省自然科学基础研究计划面上项目(2021JM-473);肺肠纤维化的中医药防治研究创新团队资助项目(2013-CXTD-01)。

摘　　要：目的:运用蛋白组学的方法研究麦冬皂苷D对小鼠肺纤维化的影响及其机制。方法:将C56BL/6小鼠按体质量随机分为正常组、模型组、阳性对照组和麦冬皂苷D组。除正常组外,其余各组小鼠气管滴注盐酸博来霉素5 mg·kg^(-1),24 h后阳性对照组给予强的松(5 mg·kg^(-1))灌胃,麦冬皂苷D组给予麦冬皂苷D(10 mg·kg^(-1))灌胃,连续给药14 d。通过显微CT观察肺组织影像学改变,染色观察小鼠肺纤维化程度,运用DIA定量蛋白组学技术检测肺组织中的差异蛋白,并分析这些差异蛋白富集的主要通路。运用Western blot(WB)对肺组织中的炎症相关蛋白进行验证。结果:与模型组比较,麦冬皂苷D给药后可显著改善小鼠肺纤维化的病理状态,CT表明小鼠肺实变减轻,染色提示肺胶原纤维化减少,肺组织中α-平滑肌肌动蛋白(α-SMA)阳性表达减轻。蛋白组学发现,相对于模型组,麦冬皂苷D组肺组织共有65个差异蛋白,其中上调蛋白46个、下调蛋白19个。GO功能富集发现,麦冬皂苷D调控的差异蛋白参与了生物过程、分子功能和细胞组分,主要富集于对压力的反应(response to stress)、碳水化合物衍生物结合(carbohydrate derivative binding)和肌原纤维(myofibril)。KEGG分析显示,麦冬皂苷D所涉及的信号通路主要包括冠状病毒疾病-新型冠状病毒肺炎(coronavirus disease-COVID-19)、补体和凝血级联(complement and coagulation cascades)、NOD样受体信号通路(NOD-like receptor signaling pathway)、肿瘤坏死因子信号通路(TNF signaling pathway)和白介素-17信号通路(IL-17 signaling pathway)等,WB结果证实麦冬皂苷D可显著降低炎症相关蛋白NOD样受体蛋白3(NLRP3)和IL-18,TNF-α,IL-6的表达(P<0.01,P<0.001)。结论:麦冬皂苷D可改善博来霉素所致的肺纤维化,其作用机制可能与减轻炎症反应、调节相关通路密切相关。Objective:To study the effect and mechanism of ophiopogonin D on pulmonary fibrosis in mice by proteomics.Methods:C56BL/6 mice were randomly divided into normal group,model group,positive control group and ophiopogonin D group according to body weight.Except the normal group,the other groups were given bleomycin hydrochloride 5 mg·kg^(-1)by tracheal drip,24 h later,the positive control group was given prednesone(5 mg·kg^(-1))by gavage,and the ophiopogonin D group was given ophiopogonin D(10 mg·kg^(-1))by gavage for 14 days.The imaging changes of lung tissues were observed by Micro-CT,and the degree of pulmonary fibrosis of mice was observed by staining.The differential proteins in lung tissues were detected by DIA quantitative proteomics technology,and the main pathways of enrichment of these differential proteins were analyzed.Western blot(WB)was used to verify the inflammation-related proteins in lung tissue.Results:Compared with the model group,the administration of ophiopogonin D could significantly improve the pathological status of pulmonary fibrosis in mice.CT showed less lung consolidation,staining indicated less lung collagen fibrosis,and the positive expression ofα-smooth muscle actin(α-SMA)in lung tissue was reduced.Proteomics showed that compared with the model group,there were 65 different proteins in the lung tissue of ophiopogonin D group,among which 46 proteins were up-regulated and 19 proteins were down-regulated.GO functional enrichment was found,differential proteins regulated by ophiopogonin D were involved in biological processes,molecular functions,and cell components,and were mainly concentrated in response to stress,carbohydrate derivative binding and myofibril.KEGG analysis showed that the signaling pathways involved in ophiopogonin D mainly included coronavirus disease-COVID-19,necroptosis,complement and coagulation cascades,NOD-like receptor signaling pathway,TNF signaling pathway and IL-17 signaling pathway.WB results showed that ophiopogonin D could significantly reduce the expre

关 键 词：肺纤维化 麦冬皂苷D 蛋白组学 

分 类 号：R965[医药卫生—药理学]