基于网络药理学探讨川芎嗪对急性坏死性胰腺炎大鼠胰腺组织的保护机制  

Protective mechanism of tetramethylpyrazine on pancreatic tissue in rats with acute necrotizing pancreatitis based on network pharmacology

作　　者：陈杰[1,2] 黄石飞[1] 韩业红 Chen Jie;Huang Shifei;Han Yehong(Department of Surgery,Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou 310021,China;Department of Emergency,Zhejiang Provincial People′s Hospital,Hangzhou 310014,China)

机构地区：[1]杭州市中医院外一科,杭州310021 [2]浙江省人民医院急诊医学科,杭州310014

出　　处：《中华胰腺病杂志》2022年第3期171-177,共7页Chinese Journal of Pancreatology

基　　金：浙江省中医药科学研究基金(2020ZB030)。

摘　　要：目的基于网络药理学探讨川芎嗪治疗急性坏死性胰腺炎(ANP)大鼠的核心靶点及其潜在的分子机制。方法通过中药系统药理学分析平台(TCMSP)及人类疾病信息相关数据库(CTD、DisGeNET、GeneCards、OMIM)筛选出川芎嗪作用靶点与ANP相关靶点,利用Uniprot数据库进行交集,导入STRING数据库构建蛋白质-蛋白质相互作用(PPI)网络,再导入Cytoscape软件进一步分析,利用cytoHubba插件得到关键靶点。对关键靶点进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,最后通过PyMol和AutoDockTools软件进行分子对接。将30只雄性SD大鼠随机分为对照组、ANP组和川芎嗪治疗组(川芎嗪组)。采用胰胆管逆行注射4%牛磺胆酸钠建立ANP大鼠模型,川芎嗪组在制模后经腹腔注射10 ml/kg川芎嗪注射液。造模后12 h,取胰腺组织,常规行病理学检查,免疫组织化学染色观察关键靶点在胰腺组织中的蛋白表达;眼眶取血,采用酶联免疫吸附测定法(ELISA)测定血清IL-6和肿瘤坏死因子-α(TNF-α)水平。结果药物平台筛选出137个川芎嗪作用靶点,疾病数据库筛选出513个ANP相关靶点,通过交集得到的25个靶点,最终获得白蛋白(ALB)、表皮生长因子受体(EGFR)、胱天蛋白酶3(CASP3)、丝裂原活化蛋白激酶1(MAPK1)和B细胞淋巴瘤样蛋白1(BCL2L1)共5个关键靶点。GO功能富集分析生物学过程主要涉及生殖结构、系统发育,对抗生素、化学应激和活性氧的反应等,细胞组成主要为囊泡腔、膜筏、膜微区和分泌颗粒腔等靶蛋白,分子功能主要包括SH2域、磷酸酪氨酸残基、蛋白酶结合及蛋白酪氨酸激酶和核受体活性等;KEGG通路富集分析主要为Ras信号通路、PI3K-Akt信号通路、铂类药物耐药、磷脂酶D信号通路和EGFR酪氨酸激酶抑制剂耐药等。5个关键靶点分子对接的平均结合能为-4.20 kcal/mol。胰腺组织病理学检查结果示ANP组大鼠�Objective To explore the core targets and potential molecular mechanisms of tetramethylpyrazine in the treatment of rats with acute necrotizing pancreatitis(ANP)based on network pharmacology.Methods The related co-targets of tetramethylpyrazine and ANP were screened out by traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)and human disease information-related databases(CTD,DisGeNET,GeneCards,OMIM);Uniprot data were used to co-link and put into the STRING database to build protein-protein interaction(PPI)networks;the Cytoscape software was used for further analysis and the key targets were obtained by using the cytoHubba plug-in.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on these key targets,and finally the molecular docking models were constructed by using PyMol and AutoDockTools software.30 male SD rats were randomly divided into the control group,ANP group,and tetramethylpyrazine treatment group(tetramethylpyrazine group).ANP rats were induced by retrograde infusion of 4%sodium taurocholate into the biliary-pancreatic duct,and the tetramethylpyrazine group rats were injected with 10 ml/kg tetramethylpyrazine through the abdominal cavity after ANP was induced.After 12 h,pancreatic tissue was taken,a pathological examination was performed routinely,and immunohistochemical staining was performed to observe the protein expression of key targets in pancreatic tissue.Blood was taken from orbits,and then the serum IL-6 and tumor necrosis factor-α(TNF-α)levels were measured by enzyme-linked immunosorbent assay(ELISA).Results The drug platform screened 137 tetramethylpyrazine action targets,and the disease database screened out 513 ANP-related targets;then 25 targets were obtained through intersection,finally resulting in a total of 5 key targets:albumin(ALB),epidermal growth factor receptor(EGFR),caspase 3(CASP3),mitogen-activated protein kinase 1(MAPK1)and B-cell lymphoma-2-like protein 1(BCL2L1).GO functional enrichment a

关 键 词：胰腺炎 急性坏死性 川芎嗪 药理学 计算生物学 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]