基于网络药理学探讨大黄保护脓毒症心肌损伤的机制研究  

Protective effect of emodin on lipopolysaccharide-induced myocardial injury based on network pharmacology

作　　者：刘艳如 刘寒琼 施良[3] 蔺博涵 任国庆 蔡华忠 张建国 孙文文[6] 宋旭 李涛[4] 刘伟[4] 张浩 Liu Yanru;Liu Hanqiong;Shi Liang;Lin Bohan;Ren Guoqing;Cai Huazhong;Zhang Jianguo;Sun Wenwen;Song Xu;Li Tao;Liu Wei;Zhang Hao(Department of Emergency,Shenzhen Third People′s Hospital,Shenzhen 518112,China)

机构地区：[1]深圳市第三人民医院急诊科,广东深圳518112 [2]张家港市第一人民医院急诊科,江苏苏州215600 [3]江苏大学附属医院急诊科,江苏镇江212000 [4]南京医科大学南京第一医院核医学科,江苏南京210006 [5]常州市妇幼保健院重症医学科,常州医学中心,南京医科大学,江苏常州213004 [6]江苏大学医学院,江苏镇江212013

出　　处：《中国急救医学》2023年第11期898-904,共7页Chinese Journal of Critical Care Medicine

基　　金：镇江市科技创新项目(SH2022042;SH202268)。

摘　　要：目的 研究大黄对脓毒症心肌损伤和线粒体功能障碍的保护作用。方法 网络药理学方法预测大黄与脓毒症心肌损伤的相关靶点,进行网络构建和蛋白质-蛋白质互作(PPI)分析,功能富集分析,构建活性成分-靶点-通路-疾病复方网络图。构建脂多糖(LPS)诱导的心肌损伤细胞模型,并将其分为正常对照组、LPS处理组和LPS+大黄组(5μmol/L、10μmol/L、15μmol/L)三组。CCK-8试剂盒检测细胞增殖,流式细胞术分析细胞凋亡,MitoSOX线粒体超氧化物红色荧光探针测定线粒体活性氧(ROS)含量,线粒体膜电位荧光探针JC-10检测线粒体膜电位水平,荧光探针Calcein AM检测线粒体通透性转换孔开放程度,透射电镜分析线粒体超微结构,蛋白质印迹(Western Blot)法测定相关蛋白表达水平。结果 (1)网络药理学结果表明,共有21个靶点与大黄治疗脓毒症心肌损伤相关。PPI构建网络显示,细胞肿瘤抗原p53(TP53)、胱天蛋白酶3(CASP3)、白细胞介素-1β(IL-1β)、髓细胞增生原癌基因(MYC)、肿瘤坏死因子(TNF)、凋亡蛋白(BAX)和抗凋亡蛋白(Bcl-2)是关键靶点。基因本体(GO)和京都基因与基因组百科全书(KEGG)分析表明,磷酯酰肌醇3激酶/蛋白激酶B(PI3K/AKT)通路在大黄调控脓毒症心肌损伤过程中起重要作用。(2)与对照组比较,LPS处理组细胞增殖能力下降且心肌细胞凋亡增加,线粒体ROS含量增加,线粒体膜电位降低,线粒体通透性转换孔开放,线粒体超微结构破坏。大黄可逆转LPS对心肌细胞的上述损伤效应。(3)在LPS诱导的脓毒症心肌损伤细胞模型中,磷酸化磷酯酰肌醇3激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)、CASP3和BAX的蛋白表达水平增加,而Bcl-2的蛋白表达水平降低。大黄可逆转上述表达,以10μmol/L效果最显著。结论 大黄通过PI3K/AKT通路调控心肌细胞线粒体氧化应激,减少细胞凋亡,降低脓毒症引起的心肌损伤。Objective To study the protective effect of emodin on myocardial injury and mitochondrial dysfunction in sepsis.Methods Network pharmacology predicted the relevant targets of emodin and myocardial injury in sepsis.We performed network construction,protein-protein interaction(PPI)analysis,functional enrichment analysis,and constructed active ingredient-target-pathway-disease compound network diagram.At the same time,the lipopolysaccharide(LPS)-induced myocardial injury models were established and divided into three groups:control group,LPS group and LPS+emodin group(5μmol/L,10μmol/L,15μmol/L).CCK-8 assay was used for cell proliferation,flow cytometry for apoptosis,the fluorescent probe MitoSOX for mitochondrial ROS content,JC-10 for mitochondrial membrane potential level,fluorescent probe Calcein AM for mitochondrial permeability conversion well openness,transmission electron microscope for mitochondrial ultrastructure analysis,and Western Blot for related protein expression.Results①Network pharmacological results showed that a total of 21 targets were associated with emodin in the treatment of myocardial injury in sepsis.The PPI network showed that TP53,CASP3,interleukin-1β(IL-1β),myelocytomatosis proto-oncogene(MYC),tumor necrosis factor(TNF),BAX and Bcl-2 were key targets.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)analysis showed that the phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)pathway played an important role in the regulation of sepsis-induced myocardial injury by emodin.②Compared with the control group,the cell proliferation ability decreased,cardiomyocyte apoptosis increased,mitochondrial ROS increased,mitochondrial membrane potential decreased,mitochondrial permeability conversion pores opened,and mitochondrial ultrastructure was destroyed in LPS-treated group.Emodin can reverse the above-mentioned damaging effects of LPS on cardiomyocytes.③In the LPS-induced sepsis myocardial injury models,the protein expression levels of phosphorylated phosphoinositide 3�

关 键 词：大黄 网络药理学 脓毒症 心肌损伤 线粒体 机制 

分 类 号：R285[医药卫生—中药学]