基于网络药理学探讨补肺通痹汤治疗系统性红斑狼疮肺间质病变的作用机制及关键通路的验证  

The mechanism of Bufei Tongbi Decoction in treatment of systemic lupus erythematosus interstitial lung disease based on network pharmacology and verification of key regulation pathway

作　　者：武上雯 李桓[1,2] 龚晓红 陆超群 陈一鸣 李析濛 李焱[1,2] 李松伟[1,2] WU Shangwen;LI Huan;GONG Xiaohong;LU Chaoqun;CHEN Yiming;LI Ximeng;LI Yan;LI Songwei(The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450003,China;Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Province Hospital of Traditional Chinese Medicine,Zhengzhou 450003,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450003 [2]河南中医药大学 [3]河南省中医院

出　　处：《北京中医药大学学报》2024年第9期1256-1271,共16页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(No.81874465);国家中医临床研究基地科研专项(No.2021JDZX2012);河南省科技攻关项目(No.222102310392)。

摘　　要：目的探讨补肺通痹汤治疗系统性红斑狼疮肺间质病变(SLE-ILD)的作用及可能的药理机制。方法采用中药系统药理学数据库与分析平台(TCMSP)、Uniprot数据库获取补肺通痹汤的有效成分和作用靶点;基于DrugBank、DisGeNET、GeneCards、PharmGKB、OMIM和GEO数据库筛选SLE-ILD关键基因;使用Cytoscape软件,构建药物有效成分-靶点-疾病关系网络图,获取补肺通痹汤治疗SLE-ILD的有效活性成分和可能作用机制。利用基因本体(GO)功能富集及京都基因和基因组百科全书(KEGG)通路富集分析揭示相关靶基因和通路功能。以C57BL/6小鼠为正常组,通过给MRL/lpr小鼠鼻腔滴注博来霉素(5 mg/kg)造模,按照随机数字表法分为模型组、补肺通痹汤低剂量组[10.4 g/(kg·d)]、补肺通痹汤中剂量组[20.8 g/(kg·d)]、补肺通痹汤高剂量组[41.6 g/(kg·d)]、醋酸泼尼松组[3 mg/(kg·d)],连续干预28 d。使用HE、Masson染色观察小鼠肺组织病理变化,免疫组织化学法检测肺组织中转化生长因子β1(TGF-β1)与Ⅲ型胶原蛋白(Col-Ⅲ)蛋白的表达,ELISA法检测血清中白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)和白细胞介素-17(IL-17)的含量,实时荧光PCR法分析肺组织中基质金属蛋白酶-1(MMP-1)、缺氧诱导因子-1α(HIF-1α)、维甲酸相关孤儿受体γt(RORγt)、叉头框蛋白P3(FOXP3)mRNA的表达,蛋白质印迹法检测肺组织中HIF-1α、MMP-1蛋白的表达,细胞流式术检测血液中辅助性T细胞17(Th17)、调节性T细胞(Treg)的表达。结果通过筛选共获得补肺通痹汤有效成分163个、作用靶点259个,SLE-ILD疾病靶点1729个,SLE-ILD差异基因958个,药物-疾病交互靶点40个。GO功能富集及KEGG通路富集发现IL-1β、MMP-1激活的IL-17信号通路,以及IL-1β、HIF-1α激活的Th17细胞分化是主要作用途径。动物实验结果显示,补肺通痹汤可有效改善SLE-ILD模型小鼠肺间质病变程度,降低TGF-β1及Col-Ⅲ表达(P<0.01);降低ILObjective To explore the effect and possible pharmacological mechanism of Bufei Tongbi Decoction in the treatment of systemic lupus erythematosus interstitial lung disease(SLE-ILD).Methods The effective components and related targets of Bufei Tongbi Decoction were obtained using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Uniprot database.Key genes for SLE-ILD were screened based on DrugBank,DisGeNET,GeneCards,PharmGKB,OMIM,and GEO databases.Using Cytoscape software,a drug active ingredient-targetdisease relationship network diagram was constructed to obtain the effective active ingredients and possible mechanisms of action of Bufei Tongbi Decoction in the treatment of SLE-ILD.Gene ontology(GO)function enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were used to reveal related target genes and pathway functions.Taking C57BL/6 mice as normal group,MRL/lpr mice were injected with bleomycin 5 mg/kg in the nasal cavity.According to the random number table method,the mice were divided into model group,Bufei Tongbi Decoction low-dose group(10.4 g/(kg·d)),Bufei Tongbi Decoction medium-dose group(20.8 g/(kg·d)),Bufei Tongbi Decoction high-dose group(41.6 g/(kg·d))and prednisone group(3 mg/(kg·d)).The intervention lasted for 28 days.Hematein eosin and Masson staining were used to observe the pathological changes of mouse lung tissue,the expressions of transforming growth factor-β1(TGF-β1)and collagen typeⅢ(Col-Ⅲ)in lung tissue were detected by immunohistochemistry,and the expressions of interleukin-1β(IL-1β),interleukin-10(IL-10)and interleukin-17(IL-17)in serum were detected by ELISA.The mRNA expressions of matrix metallopeptidase 1(MMP-1),hypoxia inducible factor-1α(HIF-1α),retinoid-related orphan receptorγt(RORγt)and forkhead box P3(FOXP3)in lung tissue were analyzed by RT-PCR,the protein expressions of HIF-1αand MMP-1 in lung tissue were detected by Western blotting,and the expressions of T helper 17 cells(Th17)

关 键 词：网络药理学 补肺通痹汤 系统性红斑狼疮肺间质病变 实验验证 小鼠 

分 类 号：R285.5[医药卫生—中药学]