基于网络药理学整合体内实验探究四物汤单味药组分治疗胆汁淤积性肝损伤的作用与机制  

Effect and mechanism of individual components of Siwu Tang in treatment of cholestatic liver injury based on network pharmacology and experimental validation in vivo

作　　者：汪乐 李佳楠 杨洋 曲姣蓉 李晓骄阳 WANG Le;LI Jianan;YANG Yang;QU Jiaorong;LI Xiaojiaoyang(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 100029,China;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学生命科学学院,北京100029 [2]北京中医药大学中药学院,北京100029

出　　处：《中草药》2024年第5期1553-1566,共14页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(82274186)。

摘　　要：目的基于网络药理学整合体内实验对比四物汤中4种单味药(川芎Chuanxiong Rhizoma、白芍Paeoniae Radix Alba、当归Angelicae Sinensis Radix、熟地黄Rehmanniae Radix Praeparata)治疗胆汁淤积性肝损伤(cholestatic liver injury,CLI)的药效差异,并探讨各单味药的作用机制。方法利用TCMSP数据库获取川芎、白芍、当归、熟地黄的主要活性成分及作用靶点,通过GeneCards数据库获取CLI靶点,使用Cyctoscape 3.10.1软件分析并构建“单味药-活性成分-作用靶点”网络,借助在线网站获取各单味药与CLI之间的交集靶点,并绘制韦恩图。将靶点基因导入DAVID数据库进行基因富集分析。构建胆总管结扎(bile duct ligation,BDL)动物模型,给予各单味药水提物后,检测血清肝功能相关生化指标,并对肝组织进行病理染色对比药效作用。同时结合qRT-PCR检测CLI相关靶点的基因表达。结果网络药理学结果显示,四物汤中川芎、白芍、当归、熟地黄与CLI之间均存在3个交集靶点[核受体亚家族3C组成员2(nuclear receptor subfamily 3 group C member 2,Nr3c2)、类视黄醇X受体(retinoid X receptor alpha,Rxra)、前列腺素内过氧化物合成酶2(prostaglandin-endoperoxide synthase 2,Ptgs2)]。通过体内验证实验表明白芍、川芎、当归、熟地黄水提物均可显著改善CLI以及相关的肝纤维化情况,且均能下调上述3个共同靶点表达(P<0.05、0.01、0.001)。此外,白芍、川芎的体内抗CLI疗效优于当归、熟地黄提取物。结果还显示白芍与CLI有18个、川芎与CLI有4个与其他单味药不同的独立交集靶点,而当归、熟地黄与CLI之间不存在独立的交集靶点。进一步生信分析发现,白芍作用靶点基因富集于血管生成、肿瘤坏死因子(tumor necrosis factor,TNF)信号通路;川芎作用靶点富集于血管内皮、血管生成、血管生成因子(vascular endothelial growth factor,VEGF)通路。进一步验证结果显示白芍通过下调肝脏中Objective To compare the therapeutic effects of the individual components of Siwu Tang(四物汤,SWT)including Chuanxiong(Chuanxiong Rhizoma),Baishao(Paeoniae Radix Alba),Danggui(Angelicae Sinensis Radix),Shudihuang(Rehmanniae Radix Praeparata)in the treatment of cholestatic liver injury(CLI)and explored the related mechanisms based on network pharmacology and experimental validation in vivo.Methods The main active ingredients and targets of Chuanxiong Rhizoma,Paeoniae Radix Alba,Angelicae Sinensis Radix and Rehmanniae Radix Praeparata were obtained by TCMSP database.CLI targets were obtained by GeneCards database and“monomeric herb-active ingredient-target”network was analyzed and constructed by Cytoscape 3.10.1 software.Intersection targets between each monomeric herb and CLI were obtained using online resources,and Venn diagram was generated.The target genes were imported into DAVID database for gene enrichment analysis.Bile duct ligation(BDL)model was conducted,after administering the water extracts of each individual herb,serum biochemistry markers related to liver function were measured,and pathological staining was performed on liver tissue to compare the pharmacological effects.Moreover,qRT-PCR was employed to detect the gene expressions of CLI related targets.Results The results of network pharmacology revealed that Chuanxiong Rhizoma,Paeoniae Radix Alba,Angelicae Sinensis Radix and Rehmanniae Radix Praeparata in SWT shared three intersecting targets[nuclear receptor subfamily 3 group C member 2(Nr3c2),retinoid X receptor alpha(Rxra)and prostaglandinendoperoxide synthase 2(Ptgs2)]with CLI.In vivo validation experiments showed that water extracts of Chuanxiong Rhizoma,Paeoniae Radix Alba,Angelicae Sinensis Radix and Rehmanniae Radix Praeparata significantly improved CLI and related liver fibrosis,and downregulated the expressions of the three common targets mentioned above(P<0.05,0.01,0.001).In addition,the in vivo anti CLI efficacy of Paeoniae Radix Alba and Chuanxiong Rhizoma were better than that e

关 键 词：网络药理学 四物汤 白芍 川芎 当归 熟地黄 胆汁淤积性肝损伤 肝纤维化 肿瘤坏死因子通路 血管生成因子通路 氧化芍药苷 洋川芎内酯A 阿魏酸 5-羟甲基糠醛 

分 类 号：R285.5[医药卫生—中药学]