牛舌草水提物调控氧化磷酸化、糖酵解/糖异生、钠尿肽系统改善维拉帕米诱导的斑马鱼心力衰竭  

Effect of aqueous extract of Anchusa italica on heart failure induced by verapamil in zebrafish by regulating oxidative phosphorylation,glycolysis/gluconogenesis and natriuretic peptide system

作　　者：铁晓璐 霍仕霞 李治建 祖力皮卡尔·吾斯曼 张云 TIE Xiaolu;HUO Shixia;LI Zhijian;ZULIPIKAER Wusiman;ZHANG Yun(College of Pharmacy,Xinjiang Medical University,Urumqi 830011,China;Uygur Medical Hospital of Xinjiang Uygur Autonomous Region,Urumqi Xinjiang Uygur Autonomous Region 830049,China;Key Laboratory of Evidence-Based and Translation,Xinjiang Hospital preparation of Traditional Chinese Medicine.,Urumqi Xinjiang Uygur Autonomous Region 830049,China;Biology Institute,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250103,China)

机构地区：[1]新疆医科大学药学院,新疆乌鲁木齐830011 [2]新疆维吾尔自治区维吾尔医医院,新疆乌鲁木齐830049 [3]新疆中药医院制剂循证与转化重点实验室,新疆乌鲁木齐830049 [4]齐鲁工业大学(山东省科学院),山东省科学院生物研究所,山东济南250103

出　　处：《药物评价研究》2025年第3期627-637,共11页Drug Evaluation Research

基　　金：中央引导地方科技发展专项资金项目(ZYYD2024JD06);天山创新团队项目(2024D14018);新疆维吾尔自治区重点实验室开放课题(KLUMH-202404)。

摘　　要：目的探讨牛舌草水提物(AIAE)抗心力衰竭作用及作用机制。方法以AB野生型及Tg(cmlc2:EGFP)斑马鱼为研究对象,设置对照组、模型(200μmol∙L^(−1)盐酸维拉帕米)组、地高辛(阳性药,10μg·mL^(−1))组、AIAE(0.1、0.5、2.5 mg·mL^(−1))组。给药后检测各组斑马鱼心包面积、静脉淤血面积、静脉窦-动脉球(SV-BA)距离、心率、短轴缩短率(FS)、射血分数(LVEF)、血流速度变化;吖啶橙染色观察心肌细胞凋亡,苏木素-伊红(HE)染色观察心脏组织病理变化;试剂盒法检测钠尿肽系统中N末端脑钠肽前体(NT-proBNP)含量;实时荧光定量PCR(qRT-PCR)法检测脑钠肽(bnp)及心钠肽(anp)mRNA表达水平。通过转录组测序(RNA-seq)预测AIAE抗心力衰竭的相关信号通路,并利用qRT-PCR检测信号通路相关基因的表达水平。结果与模型组相比,AIAE组斑马鱼心包面积、静脉淤血面积、SV-BA距离均显著减小(P<0.05、0.01、0.001),心肌细胞凋亡数减少,心率、FS、LVEF、血流速度显著升高(P<0.05、0.01、0.001),心脏壁厚度有不同程度恢复,NT-proBNP含量、bnp、anp mRNA表达水平显著降低(P<0.05、0.01、0.001)。RNA-seq结果提示AIAE可能通过调控氧化磷酸化、糖酵解/糖异生等能量代谢相关通路发挥作用。qRT-PCR验证结果表明AIAE可调控上述通路cox6a1、pck1等基因的异常表达(P<0.05、0.01、0.001)。结论AIAE通过调节氧化磷酸化、糖酵解/糖异生等信号通路,抑制钠尿肽系统激活,抑制心肌细胞凋亡发挥抗心力衰竭作用。Objective To investigate the effect and possible mechanism of aqueous extract of Anchusa italica(AIAE)on heart failure in zebrafish.Methods Zebrafish of AB wild type and Tg(cmlc2:EGFP)were used as research subjects.The control group,model group(200μmol∙L^(−1)verapamil hydrochloride),digoxin group(positive drug,10μg·mL^(−1)),and AIAE groups(0.1,0.5,2.5 mg·mL^(−1))were set up.After administration,the pericardial area,venous congestion area,distance between sinus venosus and bulbus arteriosus(SV-BA),heart rate,fractional shortening(FS),left ventricular ejection fraction(LVEF),and blood flow velocity changes of zebrafish in each group were detected.Acridine orange staining was used to observe myocardial cell apoptosis,and hematoxylin-eosin(HE)staining was used to observe pathological changes in cardiac tissue.The content of N-terminal pro-brain natriuretic peptide(NT-proBNP)in the natriuretic peptide system was detected by kit method.The mRNA expression levels of brain natriuretic peptide(bnp)and atrial natriuretic peptide(anp)were detected by real-time fluorescence quantitative PCR(qRT-PCR).The related signaling pathways of AIAE in anti-heart failure were predicted by transcriptome sequencing(RNA-seq),and the expression levels of related genes in the signaling pathways were detected by qRT-PCR.Results Compared with the model group,the pericardial area,venous congestion area,SV-BA distance,NT-proBNP content,bnp and anp mRNA expression levels were significantly reduced(P<0.05,0.01,0.001),the number of cardiomyocyte apoptosis was reduced,the heart rate,FS,LVEF and blood flow velocity were significantly increased(P<0.05,0.01,0.001),and the heart wall thickness was restored to different degrees in the AIAE groups.RNA-seq results suggested that AIAE may play a role in regulating oxidative phosphorylation,glycolysis/gluconeogenesis and other energy metabolism-related pathways.qRT-PCR results showed that AIAE could regulate the abnormal expression of cox6a1,pck1 and other genes in this pathway(P<0.05,0.01,0.001)

关 键 词：牛舌草 心力衰竭 斑马鱼 氧化磷酸化 糖酵解/糖异生 钠尿肽 

分 类 号：R285.5[医药卫生—中药学]