中国中药杂志

2022, v.47(18) 4996-5007

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

基于网络药理学及实验验证的“麻黄-葶苈子”药对治疗支气管哮喘的作用研究
Mechanism of "Ephedrae Herba-Descurainiae Semen Lepidii Semen" combination in treatment of bronchial asthma based on network pharmacology and experimental verification

张贝贝;曾梦楠;张钦钦;王茹;贾菊芳;郭彭莉;刘萌;冯卫生;郑晓珂;
ZHANG Bei-bei;ZENG Meng-nan;ZHANG Qin-qin;WANG Ru;JIA Ju-fang;GUO Peng-li;LIU Meng;FENG Wei-sheng;ZHENG Xiao-ke;School of Pharmacy,Henan University of Chinese Medicine;Engineering and Technology Center for Chinese Medicine Development of Henan Province;Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province and Education Ministry of P.R.China,Henan University of Chinese Medicine;

摘要(Abstract):

网络药理学结合体内实验探究“麻黄-葶苈子”药对治疗哮喘的作用机制,为临床用药提供理论依据。首先,利用网络药理学预测“麻黄-葶苈子”治疗哮喘的潜在靶点,构建“中药-活性成分-靶点-通路-疾病”的关系网络,并对潜在靶点进行Gene Oncology (GO)功能分析和Kyoto Encyclopedia of Genes and Genomes (KEGG)信号通路富集分析,分子对接模拟关键候选活性成分与核心基因的结合活性。然后,通过腹腔注射卵白蛋白(ovalbumin, OVA)致敏液和雾化激发构建OVA哮喘大鼠模型,分为正常组、模型组、地塞米松组(dexamethasone, DEX, 0.075 mg·kg(-1))和“麻黄-葶苈子”药对组(MT,配伍比例1∶1.5),苏木素-伊红(HE)、马松(Masson)和过碘酸雪夫(PAS)染色观察“麻黄-葶苈子”对哮喘大鼠肺脏和气管病理变化及杯状细胞增生情况的影响,酶联免疫吸附(ELISA)法检测大鼠血清中转化生长因子β1(TGF-β1)、白细胞介素6(IL6)和白细胞介素10(IL10)水平,进而通过实时荧光定量PCR(qRT-PCR)和Western blot法检测网络药理学预测的核心基因丝裂原活化蛋白激酶8(MAPK8)、cyclin D1(CCND1)、表皮生长因子受体(EGFR)、IL6、磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(Akt)的mRNA和蛋白表达水平。网络药理学分析表明“麻黄-葶苈子”治疗哮喘的潜在核心基因为MAPK8、CCND1、IL6和EGFR,并且可能与PI3K/Akt信号通路相关,“麻黄-葶苈子”药对中活性成分槲皮素和β-谷甾醇作用于哮喘的靶点较多,且分子对接结果显示槲皮素和β-谷甾醇与MAPK、PI3K和Akt具有较好的结合活性;体内实验结果表明“麻黄-葶苈子”可有效改善OVA哮喘大鼠症状,改善肺部组织病理变化,减少杯状细胞产生,减轻哮喘大鼠炎症反应,抑制MAPK8、CCND1、EGFR和IL6的表达,调节PI3K/Akt信号通路。因此,推测“麻黄-葶苈子”可能通过调节PI3K/Akt信号通路改善大鼠哮喘症状,抑制哮喘大鼠炎症反应,槲皮素和β-谷甾醇可能是“麻黄-葶苈子”治疗哮喘的候选活性成分。
This study aims to investigate mechanism of "Ephedrae Herba-Descurainiae Semen Lepidii Semen" combination(MT) in the treatment of bronchial asthma based on network pharmacology and in vivo experiment, which is expected to lay a theoretical basis for clinical application of the combination. First, the potential targets of MT in the treatment of bronchial asthma were predicted based on network pharmacology, and the "Chinese medicine-active component-target-pathway-disease" network was constructed, followed by Gene Oncology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the potential targets. Molecular docking was used to determine the binding activity of key candidate active components to hub genes. Ovalbumin(OVA, intraperitoneal injection for sensitization and nebulization for excitation) was used to induce bronchial asthma in rats. Rats were classified into control group(CON), model group(M), dexamethasone group(DEX, 0.075 mg·kg(-1))和“麻黄-葶苈子”药对组(MT,配伍比例1∶1.5),苏木素-伊红(HE)、马松(Masson)和过碘酸雪夫(PAS)染色观察“麻黄-葶苈子”对哮喘大鼠肺脏和气管病理变化及杯状细胞增生情况的影响,酶联免疫吸附(ELISA)法检测大鼠血清中转化生长因子β1(TGF-β1)、白细胞介素6(IL6)和白细胞介素10(IL10)水平,进而通过实时荧光定量PCR(qRT-PCR)和Western blot法检测网络药理学预测的核心基因丝裂原活化蛋白激酶8(MAPK8)、cyclin D1(CCND1)、表皮生长因子受体(EGFR)、IL6、磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(Akt)的mRNA和蛋白表达水平。网络药理学分析表明“麻黄-葶苈子”治疗哮喘的潜在核心基因为MAPK8、CCND1、IL6和EGFR,并且可能与PI3K/Akt信号通路相关,“麻黄-葶苈子”药对中活性成分槲皮素和β-谷甾醇作用于哮喘的靶点较多,且分子对接结果显示槲皮素和β-谷甾醇与MAPK、PI3K和Akt具有较好的结合活性;体内实验结果表明“麻黄-葶苈子”可有效改善OVA哮喘大鼠症状,改善肺部组织病理变化,减少杯状细胞产生,减轻哮喘大鼠炎症反应,抑制MAPK8、CCND1、EGFR和IL6的表达,调节PI3K/Akt信号通路。因此,推测“麻黄-葶苈子”可能通过调节PI3K/Akt信号通路改善大鼠哮喘症状,抑制哮喘大鼠炎症反应,槲皮素和β-谷甾醇可能是“麻黄-葶苈子”治疗哮喘的候选活性成分。
This study aims to investigate mechanism of "Ephedrae Herba-Descurainiae Semen Lepidii Semen" combination(MT) in the treatment of bronchial asthma based on network pharmacology and in vivo experiment, which is expected to lay a theoretical basis for clinical application of the combination. First, the potential targets of MT in the treatment of bronchial asthma were predicted based on network pharmacology, and the "Chinese medicine-active component-target-pathway-disease" network was constructed, followed by Gene Oncology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the potential targets. Molecular docking was used to determine the binding activity of key candidate active components to hub genes. Ovalbumin(OVA, intraperitoneal injection for sensitization and nebulization for excitation) was used to induce bronchial asthma in rats. Rats were classified into control group(CON), model group(M), dexamethasone group(DEX, 0.075 mg·kg(-1)), and MT(1∶1.5) group. Hematoxylin and eosin(HE), Masson, and periodic acid-Schiff(PAS) staining were performed to observe the effect of MT on pathological changes of lungs and trachea and goblet cell proliferation in asthma rats. The levels of transforming growth factor(TGF)-β1, interleukin(IL)6, and IL10 in rat serum were detected by enzyme-linked immunosorbent assay(ELISA), and the mRNA and protein levels of mitogen-activated protein kinase 8(MAPK8), cyclin D1(CCND1), IL6, epidermal growth factor receptor(EGFR), phosphatidylinositol 3-kinase(PI3 K), and protein kinase B(Akt) by qRT-PCR and Western blot. Network pharmacology predicted that MAPK8, CCND1, IL6, and EGFR were the potential targets of MT in the treatment of asthma, which may be related to PI3 K/Akt signaling pathway. Quercetin and β-sitosterol in MT acted on a lot of targets related to asthma, and molecular docking results showed that quercetin and β-sitosterol had strong binding activity to MAPK, PI3 K, and Akt. In vivo experiment showed that MT could effectively alleviate the symptoms of OVA-induced asthma rats, improve the pathological changes of lung tissue, reduce the production of goblet cells, inhibit the inflammatory response of asthma rats, suppress the expression of MAPK8, CCND1, IL6, and EGFR, and regulate the PI3 K/Akt signaling pathway. Therefore, MT may relieve the symptoms and inhibit inflammation of asthma rats by regulating the PI3 K/Akt signaling pathway, and quercetin and β-sitosterol are the candidate active components.

关键词(KeyWords): 麻黄;葶苈子;网络药理学;支气管哮喘
Ephedrae Herba;Descurainiae Semen Lepidii Semen;network pharmacology;bronchial asthma

Abstract:

Keywords:

基金项目(Foundation): 国家重点研发计划项目(2019YFC1708800);; 河南省高层次人才特殊支持计划“中原千人计划”-中原领军人才项目(ZYQR201810080)

作者(Authors): 张贝贝;曾梦楠;张钦钦;王茹;贾菊芳;郭彭莉;刘萌;冯卫生;郑晓珂;
ZHANG Bei-bei;ZENG Meng-nan;ZHANG Qin-qin;WANG Ru;JIA Ju-fang;GUO Peng-li;LIU Meng;FENG Wei-sheng;ZHENG Xiao-ke;School of Pharmacy,Henan University of Chinese Medicine;Engineering and Technology Center for Chinese Medicine Development of Henan Province;Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province and Education Ministry of P.R.China,Henan University of Chinese Medicine;

DOI: 10.19540/j.cnki.cjcmm.20220211.403

参考文献(References):

扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享