膈下逐瘀汤抑制铁死亡干预代谢相关脂肪性肝病的作用机制  

Mechanism of Gexia zhuyu decoction in the intervention of metabolic-associated fatty liver disease by inhibiting ferroptosis

作　　者：赵玉强 王自有 李安琪 赵佩然 王锐[2] 杨婧[1] ZHAO Yuqiang;WANG Ziyou;LI Anqi;ZHAO Peiran;WANG Rui;YANG Jing(School of Basic Medical Sciences,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China;School of Pharmacy,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学基础医学院,哈尔滨150040 [2]黑龙江中医药大学药学院,哈尔滨150040

出　　处：《中国药房》2024年第24期2977-2983,共7页China Pharmacy

基　　金：国家自然科学基金项目(No.82074271)。

摘　　要：目的基于铁死亡角度探讨膈下逐瘀汤干预代谢相关脂肪性肝病(MAFLD)的潜在作用机制。方法借助网络药理学方法,筛选膈下逐瘀汤干预MAFLD铁死亡的中心靶点,并进行基因本体、京都基因和基因组数据库富集分析和分子对接。将发育正常的受精后3d的斑马鱼幼鱼随机分为对照组、模型组(5mmol/L硫代乙酰胺)、异甘草酸镁组(阳性对照,5mg/mL)和膈下逐瘀汤低、中、高质量浓度组(20、40、80μg/mL,以生药量计),培养72h后检测其体内丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆固醇(TC)、甘油三酯(TG)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、活性氧(ROS)、铁离子(Fe^(2+))含量,观察其肝组织细胞结构和肝脏脂肪变性情况,检测其沉默信息调节因子1(SIRT1)、核转录因子红系2相关因子2(Nrf2)、谷胱甘肽过氧化物酶4(GPX4)蛋白的表达水平。结果膈下逐瘀汤潜在活性成分作用于MAFLD铁死亡的8个中心靶点包括肿瘤蛋白p53、SIRT1、Nrf2等;其富集于RNA聚合酶Ⅱ启动子转录的正/负向调控等生物过程,细胞核、细胞质等细胞成分,蛋白质结合等分子功能,以及铁死亡、癌症途径等信号通路,且可与主要活性成分紧密结合。与模型组比较,各药物组斑马鱼幼鱼体内ALT、AST、TC、TG、MDA、ROS、Fe^(2+)含量均显著降低,SOD、GSH含量均显著升高(P<0.05),其肝组织细胞病理损伤情况均有所改善、肝脏脂质蓄积均有所减少,SIRT1、Nrf2、GPX4蛋白的表达均显著上调(P<0.05)。结论膈下逐瘀汤可调节MAFLD模型斑马鱼幼鱼的脂质代谢,改善其氧化应激水平,维持其体内Fe^(2+)稳态,抑制其铁死亡进程,上述作用可能与激活SIRT1/Nrf2/GPX4轴有关。OBJECTIVE To explore the potential mechanism of action of Gexia zhuyu decoction in the intervention of metabolic-associated fatty liver disease(MAFLD)based on ferroptosis.METHODS With the help of network pharmacology,the central targets of Gexia zhuyu decoction intervening in ferroptosis of MAFLD were screened,then gene ontology,Kyoto Encyclopedia Gene and Genomes enrichment analysis and molecular docking were performed.Juvenile zebrafish with normal development at 3 d post-fertilization were randomly divided into control group,model group(5 mmol/L thioacetamide),magnesium isoglycyrrhizinate group(positive control,5 mg/mL),and Gexia zhuyu decoction low-,medium-and high-concentration groups(20,40,80μg/mL,calculated by crude drugs).After cultured for 72 h,the contents of alanine transaminase(ALT),aspartate transaminase(AST),total cholesterol(TC),triglyceride(TG),malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),reactive oxygen species(ROS)and Fe^(2+)were determined;the cellular structure of the liver tissues and hepatic steatosis were observed;the protein expression of silence information regulator 1(SIRT1),nuclear factor-erythroid 2-related factor 2(Nrf2)and glutathione peroxidase 4(GPX4)were detected.RESULTS The central targets of potential active ingredients of Gexia zhuyu decoction that act on ferroptosis in MAFLD included tumor proteins p53,SIRT1,Nrf2,etc.,which were enriched in biological processes such as positive/negative regulation of RNA polymeraseⅡpromoter transcription,cellular components such as nucleus and cytoplasm,molecular functions such as protein binding,as well as signaling pathways such as ferroptosis and the cancer pathway,and they might be tightly linked to the main active ingredients.Compared with model group,the contents of ALT,AST,TC,TG,MDA,ROS and Fe^(2+)were all decreased significantly in each administration group,while the contents of SOD and GSH were increased significantly(P<0.05);the pathological damage of liver tissue cells had improved,and the accumulation of liver

关 键 词：膈下逐瘀汤 代谢相关脂肪性肝病 铁死亡 SIRT1/Nrf2/GPX4轴 网络药理学 

分 类 号：R965[医药卫生—药理学] R285.5[医药卫生—药学]