Study on mechanism of Chenpi in treatment of anemia of chronic disease with Yigong Powder based on network pharmacology and experimental validation

HU Jie; ZHENG Qin; JIANG Yiling; BU Jinyang; WU Zhihao; SHI Ling; ZHANG Ruifeng; LUO Meihong

doi:10.16305/j.1007-1334.2025.2403006

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Study on mechanism of Chenpi in treatment of anemia of chronic disease with Yigong Powder based on network pharmacology and experimental validation

更新时间：2024-12-31

- Study on mechanism of Chenpi in treatment of anemia of chronic disease with Yigong Powder based on network pharmacology and experimental validation
- Shanghai Journal of Traditional Chinese Medicine Vol. 59, Issue 1, Pages: 56-65(2025)
- 作者机构：
  
  1.上海中医药大学附属宝山医院血液科（上海 201900）
  2.上海市嘉定区华亭镇社区卫生服务中心中医科;（上海 201816）
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2025.2403006
  CLC：
- Published：10 January 2025，
  
  Received：04 March 2024，
- 稿件说明：
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胡洁,郑秦,姜一陵,等.基于网络药理学和实验验证探讨陈皮在异功散治疗慢性病贫血中的作用机制[J].上海中医药杂志,2025,59(1):56-65.

HU Jie,ZHENG Qin,JIANG Yiling,et al.Study on mechanism of Chenpi in treatment of anemia of chronic disease with Yigong Powder based on network pharmacology and experimental validation[J].Shanghai Journal of Traditional Chinese Medicine,2025,59(1):56-65.
胡洁,郑秦,姜一陵,等.基于网络药理学和实验验证探讨陈皮在异功散治疗慢性病贫血中的作用机制[J].上海中医药杂志,2025,59(1):56-65. DOI： 10.16305/j.1007-1334.2025.2403006.

HU Jie,ZHENG Qin,JIANG Yiling,et al.Study on mechanism of Chenpi in treatment of anemia of chronic disease with Yigong Powder based on network pharmacology and experimental validation[J].Shanghai Journal of Traditional Chinese Medicine,2025,59(1):56-65. DOI： 10.16305/j.1007-1334.2025.2403006.

摘要

目的

为探讨陈皮在异功散治疗慢性病贫血（ACD）中的作用，通过网络药理学分析异功散中陈皮与四君子汤活性成分治疗ACD的不同作用靶点及信号通路，并进行动物实验明确陈皮活性成分在异功散治疗ACD中的作用机制。

方法

①网络药理学研究：基于中药系统药理学数据库与分析平台（TCMSP）、本草组鉴数据库（HERB）、草药成分目录平台（HIT）获取异功散方的组分，符合口服生物利用度（OB）≥30%、类药性（DL）≥0.18，或经瑞士生物信息研究所药代动力学（SwissADME）数据库筛选，符合吸收、分布、代谢和排泄标准的为最终获取的异功散活性成分。通过TCMSP、化合物靶点预测（Swisstarget prediction）数据库获得活性成分的作用靶点。再通过人类基因综合数据库（Genecards）、在线人类孟德尔遗传数据库（OMIM）、疾病相关基因与突变位点数据库（DisGeNETt）获取ACD的疾病靶点。借助韦恩（Venny）2.1在线平台获得药物靶点和疾病靶点的交集，运用蛋白互作关系数据库平台（STRING）和Cytoscape v3.9.1软件构建蛋白质网络，通过Cytoscape中MCC算法筛选前30位的核心靶点，核心靶点经京都基因与基因组百科全书（KEGG）进行信号通路富集分析。最终通过Cytoscape构建“异功散活性成分-ACD-靶点”网络和“核心靶点-信号通路”网络。②分子对接验证：比较陈皮区别于异功散方中其他活性成分的作用靶点，进行分子对接验证。③动物实验验证：建立ACD小鼠模型，进行药物干预，干预1周后检测比较各组白细胞（WBC）、血红蛋白（Hb）、红细胞（RBC）、血清铁蛋白（SF）、血清铁（SI）水平，苏木精-伊红（HE）染色脾组织观察病理学变化，蛋白免疫印迹（Western blot）法检测脾组织中磷酸化丝裂原活化蛋白激酶/丝裂原活化蛋白激酶（p-MEK/MEK）、磷酸化细胞外信号调节激酶（1/2）/细胞外信号调节激酶（1/2）［p-ERK（1/2）/ERK（1/2）］、磷酸化逆转录病毒癌蛋白网状内皮增生病毒癌基因同源物A/逆转录病毒癌蛋白网状内皮增生病毒癌基因同源物A（p-P65/P65）蛋白表达，验证陈皮活性成分对ACD的治疗作用及机制。

结果

①网络药理学分析结果显示，异功散活性成分为135种，作用于1 006个靶点，ACD疾病靶点1 474个，与异功散活性成分共同靶点为235个，计算筛选前30位核心靶点，构建得到“异功散活性成分-ACD-核心靶点”网络和“核心靶点-信号通路”网络，发现陈皮的活性成分柚皮素及其作用靶点丝裂原活化蛋白激酶3（MAPK3）及其作用的丝裂原活化蛋白激酶-细胞外信号调节激酶（1/2）-核因子-κB［MEK-ERK（1/2）-NF-κB］通路可能在异功散治疗ACD中发挥重要作用。②动物实验结果表明，与模型组比较，柚皮素组小鼠的外周血WBC、SF明显降低（

0.05），Hb、RBC、SI明显升高（

0.05）；Western blot检测显示，柚皮素组脾组织p-MEK/MEK、p-ERK（1/2）/ERK（1/2）、p-P65/P65表达均明显下降（

0.05）；HE染色结果提示，柚皮素组脾组织坏死区域、中性粒细胞浸润明显减少。

结论

在异功散方中，陈皮的有效成分柚皮素通过减少MAPK3蛋白活化来下调MEK-ERK（1/2）-NF-κB通路，降低ACD小鼠炎症状态，恢复RBC稳态，调节铁代谢，改善贫血。

Abstract

Objective

To investigate the mechanism by which Chenpi （Citri Reticulatae Pericarpium） contributes to Yigong Powder's treatment of anemia of chronic disease （ACD）， this study analyzes the different targets and signaling pathways of Chenpi and other active components of Sijunzi Tang in Yigong Powder using network pharmacology， and validates the role of Chenpi's active compounds in ACD treatment through animal experiments.

Methods

①Network pharmacology： Active components of Yigong Powder were obtained from the Traditional Chinese Medicine Systems Pharmacology Database （TCMSP）， HERB， and HIT databases， based on oral bioavailability （OB） ≥30% and drug-likeness （DL） ≥0.18， or by screening for absorption， distribution， metabolism， and excretion standards through the SwissADME database. The targets of the active components were obtained through the TCMSP and the SwissTargetPrediction database. The disease targets ACD were then identified using the human gene database （GeneCards）， the online mendelian inheritance in man （OMIM） database， and the disease-centric interaction database among diseases and various associated genes （DisGeNET）. Intersection targets were identified using the Venny 2.1 platform. Protein interaction networks were constructed using the STRING database and Cytoscape v3.9.1 software. The top 30 core targets were screened using the MCC algorithm， and Kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment analysis of the core targets was performed. Cytoscape was used to construct a "Yigong Powder active component-ACD-target" network and a "core target-signaling pathway" network. ②Molecular docking validation： The distinct targets of Chenpi， compared to other active components in Yigong Powder， were validated through molecular docking analysis. ③An ACD mouse model was prepared and treated with drug interventions. After one week of intervention， the levels of white blood cells （WBC）， hemoglobin （Hb）， red blood cells （RBC）， serum ferritin （SF）， and serum iron （SI） were measured and compared across groups. Histopathological changes in spleen tissue were observed using hematoxylin and eosin （HE） staining. Western blot analysis was performed to detect the expression of phosphorylated MEK（p-MEK）/MEK ， phosphorylated extracellular signal-regulated kinases 1/2 （p-ERK1/2）/ERK1/2， and phosphorylated P65 （p-P65）/P65 in spleen tissue， to verify the therapeutic effects and mechanisms of Chenpi's active components in the treatment of ACD.

Results

①Network pharmacology analysis revealed 135 active components in Yigong Powder， acting on 1，006 targets. There were 1，474 disease targets of ACD， with 235 common targets between the active components of Yigong Powder and ACD. The top 30 core targets were identified and used to construct the "Yigong Powder active component-ACD-core target" network and the "core target-signaling pathway" network. It was found that the active component of Chenpi， naringenin， and its target， mitogen-activated protein kinase 3 （MAPK3）， may play a crucial role in the treatment of ACD through the MEK-ERK1/2-NF-κB signaling pathway. ②Animal experiment results showed that， compared to the mice in model group， the naringenin-treated group mice showed significantly lower levels of WBC and SF （

0.05）， and significantly higher levels of Hb， RBC， and SI （

0.05） in peripheral blood. Western blot analysis showed a significant decrease in the expression of p-MEK/MEK， p-ERK（1/2）/ERK（1/2）， and p-P65/P65 proteins in spleen tissue （

0.05）. HE staining indicated a significant reduction in necrotic areas and neutrophil infiltration in spleen tissue.

Conclusion

In Yigong Powder， the active component of Chenpi， naringenin， reduces the activation of MAPK3 protein to downregulate the MEK-ERK1/2-NF-κB pathway， leading to a reduction in inflammation in ACD mice， correction of RBC homeostasis， regulation of iron metabolism， and improvement of anemia.

关键词

慢性病贫血异功散陈皮柚皮素网络药理学中药研究

Keywords

anemia of chronic diseaseYigong PowderCitri Reticulatae Pericarpiumnaringeninnetwork pharmacologytraditional Chinese herbal medicine research

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