天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (9): 1613-1623.doi: 10.16333/j.1001-6880.2023.9.015

• 数据研究 • 上一篇    下一篇

基于网络药理学和体外细胞实验验证探讨乳香治疗急性肾损伤的作用机制

白   娟,张   琰,洪术霞,史金平,徐丽婷,王   芳*   

  1. 西安国际医学中心医院药学部,西安 710100
  • 出版日期:2023-09-22 发布日期:2023-09-21
  • 基金资助:
    西安国际医学中心医院青年基金(2021QN006)

Mechanism of olibanum in the treatment of acute kidney injury based on network pharmacology and in vitro cell experiment verification

BAI Juan,ZHANG Yan,HONG Shu-xia,SHI Jin-ping,XU Li-ting,WANG Fang*   

  1. Department of Pharmacy,Xi′an International Medical Center Hospital,Xi′an 710100,China
  • Online:2023-09-22 Published:2023-09-21

摘要:

基于网络药理学研究乳香对急性肾损伤(acute kidney injury,AKI)疾病的作用机制,并根据结果建立人肾小管上皮细胞(HK-2)体外模型进行验证。通过TCMSP数据库和Swiss Target Prediction数据库筛选出223个药物靶点;应用OMIM、DisGeNet、DrugBank及GeneCards数据库收集到1 245个疾病靶点,通过韦恩图工具获取药物-疾病靶点72个。通过 String 数据库构建蛋白互作 PPI 网络;利用DAVID数据库进行GO分析和KEGG通路分析;通过Cytoscape 3.7.2软件构建网络关系图;通过Autodock进行分子对接。涉及Toll样受体、TNF、NF-κB等相关信号通路。基于此,通过细胞活性检测(CCK-8)法检测HK-2的增殖情况并以此确定分组,Hoechst33258荧光染色法检测细胞凋亡率,酶联免疫吸附测定(ELISA)法检测IL-1β、TNF-α的表达情况,聚合酶链式反应(Q-PCR)法检测MAPK3、PPARG mRNA相对表达。实验结果表明,在CCK-8实验与模型组相比,低、中、高剂量组的细胞活力逐渐增加(P <0.05);Hoechst33258、ELISA以及Q-PCR实验中,与模型组相比,低、中、高剂量组的细胞凋亡率、细胞炎性因子的产生能力和JAK2、STAT3蛋白分子量与mRNA相对表达均显著降低(P <0.05)。综上,乳香抗急性肾损伤具有多靶点、多途径的特点,为后续该药的临床应用提供了研究基础。

关键词: 乳香, 急性肾损伤, 网络药理学, TLR4, NF-κB

Abstract:

To study the mechanism of olibanum on acute kidney injury (AKI) based on network pharmacology,and establish an in vitro model of human renal tubular epithelial cells (HK-2) according to the results.To search 223 drug targets through the TCMSP database and the Swiss Target Prediction database,then using the OMIM,DisGeNet,DrugBank and GeneCards database to collect 1 245 disease targets,72 drug-disease targets were obtained through the Venn diagram tool.Protein interaction PPI network was constructed by String database;GO analysis and KEGG pathway analysis were performed using DAVID database;Cytoscape 3.7.2 software was used to construct the network diagram;Molecular docking was performed by Autodock.Involving Toll-like receptor、TNF、NF-kappa B and other signaling pathways.Based on this,the proliferation of HK-2 was detected by cell counting kit-8 (CCK-8) method,and the grouping was determined.The rate of cell apoptosis was detected by Hoechst33258 fluorescent staining,and the quantitative real time polymerase chain reaction (Q-PCR) was used to detect the relative expression of MAPK3 and PPARG mRNA.The experimental results showed that in CCK-8 experiment,compared with the model group,the cell viability in low,medium and high dose groups decreased gradually (P <0.05);Hoechst33258 experiment,ELISA experiment and Q-PCR experiment,compared with the model group,the cell apoptosis rate,the capacity of cellular inflammatory factors and the molecular weight and mRNA expression of MAPK3 and PPARG protein decreased significantly (P <0.05).In conclusion,olibanum has the characteristics of multi-target and multi-pathway,which provides a basis for the clinical application of this drug.

Key words: olibanum, acute kidney injury, network pharmacology, TLR4, NF-κB

中图分类号:  R285