摘要
目的通过网络药理学与动物体内实验,探究白鲜皮致急性肝损伤的机制。方法通过TCMSP、TCMIP和SwissTargetPrediction数据库得到白鲜皮的化学成分和作用靶点,GeneCards和CTD数据库获取肝损伤疾病的相关靶点。利用STRING数据库对交集靶点进行蛋白质互作网络分析并筛选出核心靶点,利用DAVID数据库完成GO功能和KEGG通路富集分析,使用Cytoscape软件构建“药物-成分-靶点”多层次关联网络图。动物实验中,通过灌胃给予小鼠92.7 g/(kg·d)白鲜皮,7 d后取材。采用苏木精-伊红(HE)、Masson、油红O染色观察肝脏病理学变化;酶联免疫吸附试验(ELISA)法检测小鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)、肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β的表达情况;实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测肝脏组织蛋白激酶B1(AKT1)、IL-6、TNF-α、肿瘤蛋白p53(TP53)、胱天蛋白酶3(CASP3)、IL-1β的m RNA表达。应用分子对接验证潜在毒性成分与靶点的结合情况。结果通过筛选共得到白鲜皮化学成分14个,预测靶点244个;与肝损伤的交集靶点202个。GO生物过程分析主要涉及基因表达正向调控、凋亡过程的负向调控等。KEGG通路富集分析主要包括癌症途径、磷脂酰肌醇3-激酶-蛋白激酶、TNF、IL-17等信号通路等。病理片结果显示,给予白鲜皮后,HE染色小鼠肝组织中可见重度出血,较大量的肝细胞坏死,核碎裂或溶解;Masson染色显示肝组织出现明显纤维化;油红O染色结果有大量脂滴生成。ELISA结果显示与空白组相比,给药组小鼠血清AST、ALT、ALP、LDH及肝脏MDA、TNF-α、IL-1β水平显著上升(P<0.05),肝脏SOD水平显著降低(P<0.05)。qRT-PCR结果显示给药组小鼠肝脏组织相关m RNA表达量均显著升高(P<0.05)。分子对接显示黄柏酮、白�
Objective To explore the mechanism of acute liver injury induced by Cortex dictamni through network pharmacology and in vivo experiment in animal.Methods The chemical constituents and targets of Cortex dictamni were retrieved from TCMSP,TCMIP and SwissTargetPrediction databases,and the related targets of liver injury diseases were identified through GeneCards and CTD databases.The protein interaction network of the intersection targets was analyzed by STRING database and the core targets were selected.The GO function and KEGG pathway enrichment analysis were completed by DAVID database,and the multi-level association network diagram of"drug-component-target"was constructed by Cytoscape software.In the animal study,Cortex dictamni was administered to mice at a dosage of 92.7 g/(kg·d)via intragastric administration,and the biological samples were collected after 7 days.The pathological changes of liver were observed by hematoxylin-eosin(HE),Masson and Oil Red O staining.The expression levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),and lactate dehydrogenase(LDH)in serum,as well as malondialdehyde(MDA),superoxide dismutase(SOD),tumor necrosis factor-α(TNF-α),and interleukin(IL)-1βin liver tissues,were quantified using enzyme-linked immunosorbent assay(ELISA).The expressions of protein kinase B1(AKT1),IL-6,TNF-α,tumor protein p53(TP53),cystatin 3(CASP3),and IL-1βmRNA in liver tissues were determined using real-time quantitative reverse transcription PCR(qRT-PCR).Molecular docking was employed to verify the binding affinity of potentially toxic components to their respective targets.Results A total of 14 chemical constituents,244 predicted targets and 202 intersection targets with liver injury were obtained.The GO biological process analysis mainly involved positive regulation of gene expression,negative regulation of apoptosis process.KEGG pathway enrichment analysis mainly included cancer pathway and PI3K-Akt,TNF,IL-17 signaling pathways.The pathological sectio
作者
胡文凯
徐晓敏
蒋鑫
沃佳美雪
贾素霞
刘树民
卢芳
HU Wenkai;XU Xiaomin;JIANG Xin;WO Jiameixue;JIA Suxia;LIU Shumin;LU Fang(Graduate School,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Institute of Traditional Chinese Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《中国药师》
CAS
2024年第7期1089-1098,共10页
China Pharmacist
基金
国家重点研发计划中医药现代化重点专项项目(2022YFC3502104)。
关键词
白鲜皮
急性肝损伤
网络药理学
关联网络图
分子对接
体内实验
黄柏酮
白鲜碱
梣酮
柠檬苦素
Cortex dictamni
Acute liver injury
Network pharmacology
Association network diagram
Molecular docking
In vivo experiments
Obacunone
Dictamnin
Fraxinellon
Limonin
