大黄抗肾脏纤维化的网络药理学及其活性成分的作用机制

周珊珊; 艾中柱; 李伟男; 巴元明

doi:10.13422/j.cnki.syfjx.20201002

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大黄抗肾脏纤维化的网络药理学及其活性成分的作用机制

数据挖掘 | 更新时间：2021-02-09

- 大黄抗肾脏纤维化的网络药理学及其活性成分的作用机制
- Active Components and Mechanism of Rhei Radix et Rhizoma on Renal Fibrosis Based on Network Pharmacology
- 中国实验方剂学杂志 2020年26卷第10期页码：163-172
- 作者机构：
  
  1.湖北中医药大学　中医临床学院，武汉　 430065
  2.湖北中医药大学　药学院，武汉　 430065
  3.湖北省中医院，武汉　 430061
  4.湖北省中医药研究院，武汉　 430074
- 作者简介：
  
  [第一作者]　周珊珊，在读博士，从事中医药防治慢性肾脏疾病研究，E-mail:zhouss2009@126.com
  *巴元明，硕士，教授，主任医师，从事中医药防治慢性肾脏疾病研究，E-mail: bayuanming@126.com
- 基金信息：
  
  武汉市科技计划项目(2017060201010223)
- DOI：10.13422/j.cnki.syfjx.20201002
  中图分类号： R2-0; R22; R285.5
- 收稿日期：2019-11-06，
  
  网络出版日期：2020-02-10，
  
  纸质出版日期：2020-05-20
- 稿件说明：
移动端阅览
周珊珊, 艾中柱, 李伟男, 等. 大黄抗肾脏纤维化的网络药理学及其活性成分的作用机制[J]. 中国实验方剂学杂志, 2020,26(10):163-172.

Shan-shan ZHOU, Zhong-zhu AI, Wei-nan LI, et al. Active Components and Mechanism of Rhei Radix et Rhizoma on Renal Fibrosis Based on Network Pharmacology[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(10): 163-172.
周珊珊, 艾中柱, 李伟男, 等. 大黄抗肾脏纤维化的网络药理学及其活性成分的作用机制[J]. 中国实验方剂学杂志, 2020,26(10):163-172. DOI： 10.13422/j.cnki.syfjx.20201002.

Shan-shan ZHOU, Zhong-zhu AI, Wei-nan LI, et al. Active Components and Mechanism of Rhei Radix et Rhizoma on Renal Fibrosis Based on Network Pharmacology[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(10): 163-172. DOI： 10.13422/j.cnki.syfjx.20201002.

摘要

目的：

通过网络药理学方法，预测中药大黄抗肾脏纤维化的活性成分、潜在靶点及通路，选取活性成分，对筛选出的靶基因进行体外实验验证。

方法：

检索中药系统药理学数据库与分析平台(TCMSP)与中药分子机制分析综合数据库(TCMID)获取并筛选大黄主要活性成分，利用相似度系综算法数据库(SEA)，瑞士生物信息学研究所(SIB)，GeneCards数据库预测和筛选大黄抗肾脏纤维化的潜在作用靶点。采用String Version 10.5数据库构建靶点蛋白相互作用(PPI)网络；运用David 6.8软件对关键靶点进行基因本体论(GO)富集分析、京都基因与基因组百科全书(KEGG)通路分析。采用Cytoscape Version 3.6.0软件对关键蛋白相互作用网络、活性成分-关键靶点网络、活性成分-靶点-信号通路网络进行可视化分析。结合Malarcards数据库筛选出与肾脏纤维化高关联性信号通路。进一步采用细胞实验验证：选取HK-2细胞，利用转化生长因子-

(TGF-

)诱导细胞纤维化模型，予大黄酸干预细胞48 h，蛋白免疫印迹法(Western blot)检测缺氧诱导因子1

(HIF-1

)，血管内皮生长因子(VEGF)，血小板衍生生长因子受体-

(PDGFR-

)，免疫荧光检测E-钙黏蛋白(E-cadherin)，

-平滑肌肌动蛋白(

-SMA)蛋白表达，流式细胞术检测细胞凋亡。

结果：

筛选得到大黄活性成分17个，大黄抗肾脏纤维化潜在靶点424个，关键靶点5个，依次为蛋白激酶B(Akt)1，丝裂原激活的蛋白激酶(MAPK)3，EGFR，白细胞介素-6(IL-6)，VEGFA；GO富集生物学过程主要涉及信号转导、细胞增殖、凋亡等生物过程；KEGG通路富集结果发现磷脂酰肌醇3-激酶(PI3K)/Akt，HIF-1

，VEGF，叉头转录因子(FoxO)等通路与大黄抗肾脏纤维化作用机制相关。体外实验验证表明，大黄酸抑制E-cadherin，

-SMA，HIF-1

，VEGF，PDGFR-

的表达水平，同时抑制肾小管上皮细胞凋亡，验证了网络药理学部分预测结果。

结论：

本研究体现了大黄多成分、多靶点、多途径的作用特点，其抗肾脏纤维化的作用机制可能与其抑制HIF-1

/VEGF/PDGFR-

信号转导途径，抑制细胞凋亡及肾小管上皮细胞上皮间质转化(EMT)有关。

Abstract

Objective:

To explore the active components

potential targets and signaling pathways of Rhei Radix et Rhizoma in the treatment of renal fibrosis based on the network pharmacology method

and then to verify the target genes

in vitro

Method:

Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Traditional Chinese Medicine Integrated Database (TCMID) were retrieved to obtain the main active ingredients of Rhei Radix et Rhizoma. The potential anti-renal fibrosis targets of Rhei Radix et Rhizoma were predicted by similarity ensemble approach (SEA)

Swiss Institute of Bioinformatics (SIB) and GeneCards Database. Target protein-protein interaction (PPI) network was constructed by using String Version 10.5 database. David 6.8 software was used for gene ontology (GO) enrichment analysis and the Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of the key targets. Cytoscape Version 3.6.0 software was used for visualized analysis of PPI network

active ingredient-key target network and the ingredient-target-signal pathway network. In combination with Malachards database

the signal pathways with high correlation with renal fibrosis were screened. Then

cell experiments were used for verification: HK-2 cells were selected to establish fibrosis model by transforming growth factor-

(TGF-

) stimulation. The cells were treated with rhein for 48 hours. Western blot assay was used to detect the protein expression level of hypoxia inducible factor-1

(HIF-1

)

vascular endothelial growth factor (VEGF)

and platelet-derived growth factor receptor-

(PDGFR-

). Protein expression levels of E-cadherin and

smooth muscle actin (

-SMA) were detected by immunofluorescence. Apoptosis was detected with flow cytometry.

Result:

Totally 17 active ingredients of Rhei Radix et Rhizoma and 424 targets of anti-renal fibrosis effect were screened out

including five key targets: protein kinase B(Akt)1

mitogen activated protein kinases 3(MAPK3)

epidermal growth factor receptor(EGFR)

interleukin(IL)-6 and VEGFA in turn. The biological process of GO enrichment mainly involved signal transduction

cell proliferation and apoptotic process. The results of KEGG pathway enrichment showed that phosphatidylinositol 3-kinase(PI3K)/Akt

HIF-1

VEGF

and forkhead transcription factor (FoxO) pathways were related to the anti-renal fibrosis mechanism of Rhei Radix et Rhizoma. Results of the

in vitro

experiment proved that rhein could inhibit the expression of E-cadherin

-SMA

HIF-1

VEGF and PDGFR-

. In addition

rhein inhibited apoptosis induced by TGF-

in HK-2 cells. Part of the prediction results of network pharmacology were verified.

Conclusion:

This study reflects the multi-component

multi-target and multi-pathway mechanism characteristics of Rhei Radix et Rhizoma. The mechanisms of its anti-renal fibrosis effects may be related to inhibiting HIF-1

/ VEGF /PDGFR-

signaling pathway

apoptosis and epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells.

关键词

Keywords

references

A LI , X ZHANG , M SHU , et al . Arctigenin suppresses renal interstitial fibrosis in a rat model of obstructive nephropathy [J]. Phytomedicine ， 2017 ， 30 ： 28 - 41 .

王亦君，冯舒涵，程锦堂，等．大黄蒽醌类化学成分和药理作用研究进展 [J]．中国实验方剂学杂志， 2018 ， 24 ( 13 )： 227 - 234 .

X DONG , J FU , X YIN , et al . Emodin: a review of its pharmacology, toxicity and pharmacokinetics [J]． Phytother Res , 2016 , 30 ( 8 ): 1207 - 1218 .

Z LU , Y ZENG , F LU , et al . Rhei Radix et Rhizoma enema attenuates renal tubulointerstitial fibrosis in 5/6 nephrectomized rats by alleviating indoxyl sulfate overload [J]． PLoS One , 2015 ， 10 ( 12 )： e0144726 .

Y BAI , Z SU , H SUN , et al . Aloe-emodin relieves high-fat diet induced QT prolongation via MiR-1 inhibition and IK1 up-regulation in rats [J]. Cell Physiol Biochem , 2017 ， 43 ( 5 )： 1961 - 1973 .

Q HUANG , G LU , H M SHEN , et al . Anti-cancer properties of anthraquinones from Rhei Radix et Rhizoma [J]. Med Res Rev ， 2007 ， 27 ( 5 )： 609 - 630 .

T CHEN , L Y ZHENG , W XIAO , et al . Emodin ameliorates high glucose induced-podocyte epithelial-mesenchymal transition in - vitro and in - vivo [J]. Cell Physiol Biochem , 2015 , 35 ( 4 ): 1425 - 1436 .

D HE , L LEE , J YANG , et al . Preventive effects and mechanisms of rhein on renal interstitial fibrosis in obstructive nephropathy [J]. Biol Pharm Bull , 2011 , 34 ( 8 ): 1216 - 1219 .

郑文江，谢富淇，黄慧婷，等．基于网络药理学的射干-麻黄药对治疗支气管哮喘的分子机制探讨 [J]．中国实验方剂学杂志， 2019 ， 25 ( 19 )： 182 - 194 .

Y ZHANG , Y LI , X MAO , et al . Thyroid hormone synthesis: a potential target of a Chinese herbal formula HaizaoYuhu Decoction acting on iodine-deficient goiter [J]. Oncotarget ， 2016 ， 7 ( 32 ): 51699 - 51712 .

S SUN , X NING , Y ZHANG , et al . Hypoxia-inducible factor-1alpha induces Twist expression in tubular epithelial cells subjected to hypoxia, leading to epithelial-to-mesenchymal transition [J]. Kidney Int ， 2009 ， 75 ： 1278 - 1287 .

沈倩，冯松杰．中医药治疗慢性肾衰竭临床研究进展 [J]．河北中医， 2014 ， 36 ( 4 ): 625 - 627 .

党珏，李燕，刘美辰，等．大黄有效成分与附子有效成分配伍在Caco-2细胞模型上的转运分析 [J]．中国实验方剂学杂志， 2018 ， 24 ( 7 )： 1 - 6 .

李妹娟，王和生，王通渤，等．制何首乌中大黄素对ApoE～ (-/-) 小鼠动脉粥样硬化模型中JAK2/STAT3通路的影响 [J]．中国实验方剂学杂志， 2018 ， 24 ( 18 )： 101 - 106 .

许科，张犁．大黄酸抗肾脏纤维化研究进展 [J]．新中医， 2012 ， 44 ( 5 )： 119 - 121 .

D HE , L LEE , J YANG , et al . Preventive effects and mechanisms of rhein on renal interstitial fibrosis in obstructive nephropathy [J]． Biol Pharm Bull , 2011 , 34 ( 8 ): 1219 - 1226 .

Y TU , L GU , D CHEN , et al . Rhein inhibits autophagy in rat renal tubular cells by regulation of AMPK/mTOR signaling [J]． Sci Rep ， 2017 ， doi: doi: 10.1038/srep43790 http://dx.doi.org/10.1038/srep43790 .

S KOBAYASHI , T J BOGGON , T DAYARAM , et al . EGFR mutation and resistance of non-small-cell lung cancer to gefitinib [J]. N Engl J Med , 2005 ， 352 ( 8 )： 786 - 792 .

F DOU , Y LIU , L LIU , et al . Aloe-emodin ameliorates renal fibrosis via inhibiting PI3K/Akt/mTOR signaling pathway in vivo and in vitro [J]． Rejuvenation Res , 2019 ， 22 ( 3 )： 218 - 229 .

T JI , S L SU , Y ZHU , et al . The mechanism of mulberry leaves against renal tubular interstitial fibrosis through ERK1/2 signaling pathway was predicted by network pharmacology and validated in human tubular epithelial cells [J]． Phytother Res ， 2019 ， 33 ( 8 )： 2044 - 2055 .

X PAN , J LI , X HE , et al . Gadolinium chloride promotes proliferation of HEK293 human embryonic kidney cells by activating EGFR/PI3K/Akt and MAPK pathways [J]. Biometals , 2019 ， 32 ( 4 )： 683 - 693 .

D F HIGGINS , K KIMURA , W M BERNHARDT , et al . Hypoxia promotes fibrogenesis in vivo via HIF-1 stimulation of epithelial-mesenchymal transition [J]． J Clin Invest , 2007 , 117 ( 12 )： 3810 - 3820 .

J HU , W WANG , F ZHANG , et al . Hypoxia inducible factor-1 α mediates the profibrotic effect of albumin in renal tubular cells [J]. Sci Rep , 2017 , 7 ( 1 ): 15878 .

M GERT ． Capillary rarefaction, hypoxia, VEGF and angiogenesis in chronic renal disease [J]． Nephrol Dial Transplant ， 2011 , 26 : 1132 - 1137 .

C JAMES . Bonner Regulation of PDGF and its receptors in fibrotic diseases [J]． Cytokine Growth Factor Rev , 2004 ， 15 ( 4 ): 255 - 273 .

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