Mechanism of Huangqi Decoction against liver fibrosis by network pharmacological analysis combined with related cell experiment

QIN Yan; LI Yajuan; WANG Qinglan

doi:10.16305/j.1007-1334.2023.2101139

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Mechanism of Huangqi Decoction against liver fibrosis by network pharmacological analysis combined with related cell experiment

更新时间：2023-11-24

- Mechanism of Huangqi Decoction against liver fibrosis by network pharmacological analysis combined with related cell experiment
- Shanghai Journal of Traditional Chinese Medicine Vol. 57, Issue 11, Pages: 67-76(2023)
- 作者机构：
  
  1.上海中医药大学交叉科学研究院（上海 201203）
  2.上海中医药大学中医学院（上海 201203）
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2023.2101139
  CLC：
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秦艳,李亚娟,王清兰.网络药理学分析结合细胞实验探讨黄芪汤抗肝纤维化的作用机制[J].上海中医药杂志,2023,57(11):67-76.

QIN Yan,LI Yajuan,WANG Qinglan.Mechanism of Huangqi Decoction against liver fibrosis by network pharmacological analysis combined with related cell experiment[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(11):67-76.
秦艳,李亚娟,王清兰.网络药理学分析结合细胞实验探讨黄芪汤抗肝纤维化的作用机制[J].上海中医药杂志,2023,57(11):67-76. DOI： 10.16305/j.1007-1334.2023.2101139.

QIN Yan,LI Yajuan,WANG Qinglan.Mechanism of Huangqi Decoction against liver fibrosis by network pharmacological analysis combined with related cell experiment[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(11):67-76. DOI： 10.16305/j.1007-1334.2023.2101139.

摘要

目的,2,基于网络药理学方法分析黄芪汤抗肝纤维化的关键分子机制，并进一步采用相关肝星状细胞（HSC）实验验证其作用机制。,方法,2,通过中药系统药理学数据库分析平台（TCMSP）获取黄芪汤中黄芪和甘草的全部化学成分，根据生物利用度（OB）≥30%和类药性（DL）≥0.18的筛选条件筛选出潜在的活性成分，同时通过化源网（Chemsrc）数据库以及检索文献补充数据库中未获得的活性成分。通过TCMSP、化合物靶点预测（Swisstarget prediction）数据库、药物银行（DrugBank）数据库获得化学成分的作用靶点，即药物靶点；再通过全基因（Gencards）数据库、在线人类孟德尔遗传（OMIM）数据库等获取肝纤维化的主要靶点，即疾病靶点；取二者交集获得黄芪汤成分抗肝纤维化的潜在作用靶点。采用富集分析（Metascape）平台进行京都基因与基因组百科全书（KEGG）数据库信号通路分析，获得黄芪汤成分调控的关键信号通路，并采用Cytoscape 3.8.1软件构建“黄芪汤抗肝纤维化靶点-信号通路”网络图。最后利用体外人肝星状细胞系人肝星状细胞LX-2（简称“LX-2细胞”）验证黄芪汤对LX-2细胞活化及关键信号通路的影响。,结果,2,通过网络药理学分析筛选出153个黄芪汤成分抗肝纤维化的潜在作用靶点。KEGG信号通路分析提示，上述靶点可能与白介素-17（IL-17）信号通路、磷脂酰肌醇-3-激酶-蛋白激酶B（PI3K-Akt）信号通路、5'-单磷酸腺苷依赖的蛋白激酶（AMPK）信号通路等密切相关。PI3K-Akt信号通路在肝纤维化发生发展，尤其是HSC活化中发挥重要作用。体外研究发现，黄芪汤可下调LX-2细胞α平滑肌肌动蛋白（α-SMA）及Ⅰ型胶原蛋白（Col-Ⅰ）基因和蛋白表达，抑制细胞活化；进一步研究发现，黄芪汤对LX-2细胞PI3K基因及蛋白表达有显著抑制作用，并可下调Akt的磷酸化水平，提示对PI3K-Akt信号通路有抑制作用。,结论,2,网络药理学分析结合细胞实验提示，黄芪汤抗肝纤维化的部分作用机制与下调PI3K-Akt信号通路、抑制肝星状细胞活化相关。

Abstract

Objective,2,To investigate the key molecular mechanism of Huangqi Decoction against liver fibrosis based on network pharmacology and hepatic stellate cells （HSC） experiment validation.,Methods,2,Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） was used to obtain all the components from Astragali Radix and Glycyrrhizae Radix et Rhizoma. Then， the potential active components were screened out with the criteria of bioavailability （OB）≥30% and drug-like （DL）≥0.18； Meanwhile， more potential active components were acquired by Chemsrc database and literature. The targets of chemical components， also called drug targets， were obtained through TCMSP database， Swisstarget prediction database and DrugBank. Then the targets of hepatic fibrosis， also called disease targets， were obtained through Gencards， OMIM. The potential targets of anti-hepatic fibrosis of Huangqi Decoction were obtained by the intersection of drug targets and disease targets. The signal pathway of Kyoto Encyclopedia of Gene and Genome （KEGG） was analyzed by enrichment analysis （Metascape） platform， and the key signal pathways of Huangqi Decoction were obtained， and the network map of “Huangqi Decoction anti-hepatic fibrosis target-signal pathway” was constructed by Cytoscape 3.8.1 software. Finally， the effects of Huangqi Decoction on the activation and key signal pathways of LX-2 cells were verified by human hepatic stellate cell LX-2 ,in vitro,.,Results,2,A total of 153 potential targets of Huangqi Decoction against liver fibrosis were screened by network pharmacological analysis. The analysis of KEGG signal pathway suggests that the above targets may be closely related to interleukin-17 （IL-17） signal pathway， phosphatidylinositol-3-kinase-protein kinase B （PI3K-Akt） signal pathway and 5-adenosine monophosphate-dependent protein kinase （AMPK） signal pathway. PI3K-Akt signaling pathway plays an important role in the occurrence and development of liver fibrosis， especially in HSC activation. ,In vitro,， Huangqi Decoction could down-regulate the gene and protein expression of α-smooth muscle actin （α-SMA） and type Ⅰ collagen （Col-Ⅰ） in human hepatic stellate cell line LX-2 cells， and inhibit cell activation. Further study showed that Huangqi Decoction could significantly inhibit the expression of PI3K gene and protein in LX-2 cells， and down-regulate the phosphorylation level of Akt， suggesting that it could inhibit the PI3K-Akt signal pathway.,Conclusion,2,Network pharmacology combined with cell experiment suggests that part of the mechanism of Huangqi Decoction in anti-hepatic fibrosis is related to down-regulating PI3K-Akt signal pathway and inhibiting the activation of hepatic stellate cells.

关键词

肝纤维化黄芪汤网络药理学肝星状细胞作用机制中药研究

Keywords

hepatic fibrosisHuangqi Decoctionnetwork pharmacologyhepatic stellate cellsmechanism of actiontraditional Chinese herbal medicine research

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