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- 北京中医药大学学报 2024年47卷第9期 页码：1256-1271
- 作者机构：
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
- 作者简介：
武上雯，女，博士，主治医师
#李松伟，男，博士，主任医师，博士生导师，主要研究方向：中医药防治风湿免疫疾病，E-mail：ryanzzdx@yeah.net
- 基金信息：国家自然科学基金面上项目(81874465);国家中医临床研究基地科研专项(2021JDZX2012);河南省科技攻关项目(222102310392)
DOI：10.3969/j.issn.1006-2157.2024.09.010
中图分类号： R285.5纸质出版日期：2024-09-30，
网络出版日期：2024-08-02，
收稿日期：2024-05-12，
- 稿件说明：
移动端阅览
武上雯, 李桓, 龚晓红, 等. 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[J]. 北京中医药大学学报, 2024,47(9):1256-1271.
WU Shangwen, LI Huan, GONG Xiaohong, et al. 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[J]. Journal of Beijing University of Traditional Chinese Medicine, 2024,47(9):1256-1271.武上雯, 李桓, 龚晓红, 等. 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[J]. 北京中医药大学学报, 2024,47(9):1256-1271. DOI： 10.3969/j.issn.1006-2157.2024.09.010.
WU Shangwen, LI Huan, GONG Xiaohong, et al. 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[J]. Journal of Beijing University of Traditional Chinese Medicine, 2024,47(9):1256-1271. DOI： 10.3969/j.issn.1006-2157.2024.09.010.
摘要
目的
2
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
2
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
2
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1 729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IL-1β、HIF-1α、IL-17的表达(
P
<
0.01)；中、高剂量
补肺通痹汤下调MMP-1、RORγt mRNA表达(
P
<
0.01)，上调IL-10与FOXP3 mRNA表达(
P
<
0.01)；各剂量补肺通痹汤可使Th17细胞比例降低，Treg细胞比例升高，下调Th17/Treg平衡状态(
P
<
0.05)，改善免疫紊乱状态。
结论
2
补肺通痹汤治疗SLE-ILD具有多靶点、多途径特点，其作用机制可能与调控Th17/Treg细胞平衡有关。
Abstract
Objective
2
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
2
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-target-disease 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
type Ⅲ(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 receptor γt (RORγt ) and forkhead box P3 (FOXP3) in lung tissue were analyzed by RT-PCR
the protein expressions of HIF-1α and MMP-1 in lung tissue were detected by Western blotting
and the expressions of T helper 17 cells (Th17) and regulatory T cells (Treg cells) in blood were detected by cytometry.
Results
2
A total of 163 effective ingredients
259 targets
1 729 SLE-ILD disease targets
958 SLE-ILD differential genes and 40 drug-disease interaction targets were obtained by screening. GO functional enrichment and KEGG pathway enrichment showed that IL-17 signaling pathway activated by IL-1β and MMP-1
and Th17 cell differentiation activated by IL-1β and HIF-1α were the main pathways. Animal experiments showed that
Bufei Tongbi
Decoction could effectively improve the degree of lung interstitial lesion and reduce the expressions of TGF-β1 and Col-Ⅲ in SLE-ILD mice (
P
<
0.01). The expressions of IL-1β
HIF-1α and IL-17 were decreased (
P
<
0.01). Medium and high doses of
Bufei Tongbi
Decoction decreased the expressions of MMP-1 and RORγt mRNA (
P
<
0.01)
and increased the expressions of IL-10 and FOXP3 mRNA (
P
<
0.01).
Bufei Tongbi
Decoction could reduce the proportion of Th17 cells
increase the proportion of Treg cells
downregulate the balance of Th17/Treg (
P
<
0.05)
and improve the immune disorder.
Conclusion
2
Bufei Tongbi
Decoction has the characteristics of multi-target and multi-pathway in treating SLE-ILD
and its mechanism may be related to the regulation of Th17/Treg
cell balance.
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
Keywords
network pharmacologyBufei Tongbi Decoctionsystemic lupus erythematosus interstitial lung diseaseexperimental verificationmice
references
TIAN JR, ZHANG DY, YAO X, et al. Global epidemiology of systemic lupus erythematosus: a comprehensive systematic analysis and modelling study[J]. Ann Rheum Dis, 2023, 82(3): 351－356.
AMARNANI R, YEOH SA, DENNENY EK, et al. Lupus and the lungs: the assessment and management of pulmonary manifestations of systemic lupus erythematosus[J]. Front Med, 2020, 7: 610257.
HA YJ, LEE YJ, KANG EH. Lung involvements in rheumatic diseases: update on the epidemiology, pathogenesis, clinical features, and treatment[J]. Biomed Res Int, 2018, 2018: 6930297.
ARINGER M, COSTENBADER K, DAIKH D, et al. 2019 European League Against Rheumatism/American College of Rheumatology classification criteria for systemic lupus erythematosus[J]. Ann Rheum Dis, 2019, 78(9): 1151－1159.
RAGHU G, ROCHWERG B, ZHANG Y, et al. An official ATS/ERS/JRS/ALAT clinical practice guideline: treatment of idiopathic pulmonary fibrosis. An update of the 2011 clinical practice guideline[J]. Am J Respir Crit Care Med, 2015, 192(2): e3－e19.
FANOURIAKIS A, KOSTOPOULOU M, ALUNNO A, et al. 2019 update of the EULAR recommendations for the management of systemic lupus erythematosus[J]. Ann Rheum Dis, 2019, 78(6): 736－745.
武上雯，李桓，郭洪涛，等. 基于真实世界数据挖掘中医药治疗不同证型系统性红斑狼疮肺间质病变的用药规律研究[J]. 中国中医基础医学杂志，2024, 30(1): 88－94.
NOGALES C, MAMDOUH ZM, LIST M, et al. Network pharmacology: curing causal mechanisms instead of treating symptoms[J]. Trends Pharmacol Sci, 2022, 43(2): 136－150.
ZHAO L, ZHANG H, LI N, et al. Network pharmacology, a promising approach to reveal the pharmacology mechanism of Chinese medicine formula[J]. J Ethnopharmacol, 2023, 309: 116306.
SZAPIEL SV, ELSON NA, FULMER JD, et al. Bleomycin-induced interstitial pulmonary disease in the nude, athymic mouse[J]. Am Rev Respir Dis, 1979, 120(4): 893－899.
ASHCROFT T, SIMPSON J M, TIMBRELL V. Simple method of estimating severity of pulmonary fibrosis on a numerical scale[J]. J Clin Pathol, 1988, 41(4): 467－470.
赵志娜，李桓，李松伟．系统性红斑狼疮肺间质病变的中医病机探析[J].风湿病与关节炎，2019，8(11)：57－59.
刘芬芬，羊维，黄琳，等. 中医学对糖皮质激素主治功效的药性认识[J]. 中华中医药杂志，2015, 30(4): 1268－1270.
BASARAN E, ÖZTÜRK AA, SENEL B, et al. Quercetin, rutin and quercetin-rutin incorporated hydroxypropyl β－cyclodextrin inclusion complexes[J]. Eur J Pharm Sci, 2022, 172: 106153.
YUAN K, ZHU QQ, LU QY, et al. Quercetin alleviates rheumatoid arthritis by inhibiting neutrophil inflammatory activities[J]. J Nutr Biochem, 2020, 84: 108454.
LI Y, YAO JY, HAN CY, et al. Quercetin, inflammation and immunity[J]. Nutrients, 2016, 8(3): 167.
薛兰，毛春迎，王慧，等. 槲皮素治疗大鼠肺纤维化的作用及相关机制研究[J]. 中国比较医学杂志，2022, 32(7): 94－100, 117.
LI XH, ZHAI YQ, XI BR, et al. Pinocembrin ameliorates skin fibrosis via inhibiting TGF-β1 signaling pathway[J]. Biomolecules, 2021, 11(8): 1240.
LIU HQ, YU H, CAO ZJ, et al. Kaempferol modulates autophagy and alleviates silica-induced pulmonary fibrosis[J]. DNA Cell Biol, 2019, 38(12): 1418－1426.
GUPTA I, ADIN SN, PANDA BP, et al. β－Carotene-production methods, biosynthesis from Phaffia rhodozyma, factors affecting its production during fermentation, pharmacological properties: a review[J]. Biotechnol Appl Biochem, 2022, 69(6): 2517－2529.
YI XYL, LI J, LIAO DZ, et al. Carrot and carotene and multiple health outcomes: an umbrella review of the evidence[J]. J Sci Food Agric, 2023, 103(5): 2251－2261.
LOPEZ-CASTEJON G, BROUGH D. Understanding the mechanism of IL-1β secretion[J]. Cytokine Growth Factor Rev, 2011, 22(4): 189－195.
RASCHI E, CHIGHIZOLA CB, CESANA L, et al. Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts[J]. Arthritis Res Ther, 2018, 20(1): 187.
SEMENZA GL. HIF－1 and mechanisms of hypoxia sensing[J]. Curr Opin Cell Biol, 2001, 13(2): 167－171.
YANG C, ZHONG ZF, WANG SP, et al. HIF－1: structure, biology and natural modulators[J]. Chin J Nat Med, 2021, 19(7): 521－527.
SHARABI A, TSOKOS GC. T cell metabolism: new insights in systemic lupus erythematosus pathogenesis and therapy[J]. Nat Rev Rheumatol, 2020, 16(2): 100－112.
YASUDA K, TAKEUCHI Y, HIROTA K. The pathogenicity of Th17 cells in autoimmune diseases[J]. Semin Immunopathol, 2019, 41(3): 283－297.
LEE GR. The balance of Th17 versus Treg cells in autoimmunity[J]. Int J Mol Sci, 2018, 19(3): 730.
PLACHOURI KM, CHOURDAKIS V, GEORGIOU S. The role of IL-17 and IL-17 receptor inhibitors in the management of generalized pustular psoriasis[J]. Drugs Today, 2019, 55(9): 587－593.
NA HS, PARK JS, CHO KH, et al. Interleukin－1－interleukin－17 signaling axis induces cartilage destruction and promotes experimental osteoarthritis[J]. Front Immunol, 2020, 11: 730.
0
浏览量
9
下载量
0
CSCD
- 网络PDF下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 地址：北京市朝阳区北三环东路11号 邮编：100029
- 联系电话：(010)53911857 Email：jbutcm@bucm.edu.cn
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies