基于斑马鱼模型研究活心丸对阿霉素心脏毒性的预防和作用机制  

Prevention and Mechanism of Huoxin Pill on Doxorubicin-Induced Cardiotoxicity Based on Zebrafish

作　　者：曾来凤 林久茂 叶锦霞 李宪美 ZENG Laifeng;LIN Jiumao;YE Jinxia;LI Xianmei(Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学中西医结合研究院,福建福州350122 [2]福建省中西医结合老年性疾病重点实验室,福建福州350122

出　　处：《福建中医药》2024年第2期36-40,共5页Fujian Journal of Traditional Chinese Medicine

基　　金：福建省自然科学基金项目(2021J05063);福建中医药大学校管课题(X2020010-重点);福建中医药大学高层次人才科研启动资金项目(X2020013-人才)。

摘　　要：目的 基于斑马鱼模型探究复方中药活心丸(HXP)对阿霉素(DOX)所致心脏毒性的预防及其作用机制。方法 选取7月龄野生型AB品系雌性斑马鱼80尾,随机分为对照组、模型组、低剂量组、高剂量组各20尾。第1~7天,低、高剂量组分别按生药量9、12μg/(g·d)予HXP药液腹腔注射,对照组和模型组按5.7μL/(g·d)予生理盐水腹腔注射;第8天,模型组及低、高剂量组按生药量20μg/(g·d)予DOX药液腹腔注射,对照组按5.7μL/(g·d)予生理盐水腹腔注射,随后在养殖系统中静养4周。给药注射后每隔7 d观察各组斑马鱼存活情况,并计算存活率。静养4周后进行麻醉、称重,取斑马鱼心脏组织,测量心室面积,计算心室面积/体质量。Masson染色观察心肌组织纤维化程度;α-actinin免疫荧光染色观察心肌排列情况;TUNEL染色观察心肌组织凋亡情况;qPCR检测心脏组织前列腺素内过氧化物合成酶(ptgs2)、花生四烯酸5脂氧合酶(alox5a)、NADPH氧化酶1(nox1)、抗氧化应激因子谷胱甘肽过氧化酶1(gpx1)、心房利钠因子(anf)、促炎因子肿瘤坏死子α(tnfα)、白细胞介素6(il6)和抑炎因子转化生长因子β1(tgfβ1b)mRNA相对表达水平。结果 与对照组比较,模型组心室面积和心室面积/体质量均明显升高(P<0.05),ptgs2、alox5a、nox1、anf、tnfα、il6 mRNA相对表达水平均明显提高(P<0.05),gpx1 mRNA相对表达水平均明显降低(P<0.05);病理可见心肌肥大、粘连、纤维化,心肌组织排列紊乱,凋亡细胞增多。与模型组比较,高剂量组心室面积和心室面积/体质量均明显降低(P<0.05),ptgs2、alox5a、nox1、anf、tnfα、il6 mRNA相对表达水平均明显降低(P<0.05),tgfβ1b、gpx1 mRNA相对表达水平均明显升高(P<0.05);病理可见心肌肥大、粘连、纤维化、心肌组织排列紊乱和细胞凋亡情况改善。结论 活心丸可有效预防阿霉素所致的斑马鱼心脏病理改变,其机制可能与下调铁死亡和炎症�Objective:To discuss the protective effect and mechanism of Huoxin pill(HXP)on doxorubicin(DOX)induced cardiotoxicity in zebrafish model.Methods:Eighty wild-type AB strain female zebrafish were randomly divided into control group,model group,low-dose group and high-dose group,with 20 zebrafish in each group.On the 1st to 7th day,the low-and high-dose groups were intraperitoneally injected with HXP solution at the dose of 9 and 12μg/(g·d),respectively,and the control and model groups were intraperitoneally injected with normal saline at the dose of 5.7μL/(g·d).On the 8th day,the model,low-dose and high-dose groups were intraperitoneally injected with DOX solution at the dose of 20μg/(g·d),and the control group was intraperitoneally injected with normal saline at the dose of 5.7μL/(g·d).The survival of zebrafish in each group was observed every 7 days after DOX injection.After 4 weeks of DOX injection,ventricular area and ventricular area/body weight were measured.Masson staining was used to observe the degree of myocardial tissue fibrosis;α-actinin immunofluorescence staining was used to observe the arrangement of myocardium;Tunel staining was used to observe the apoptosis of myocardial tissue;qPCR was used to detect the mRNA expression level of prostaglandin-endoperoxide synthase 2(ptgs2),arachidonate 5-lipoxygenase(alox5a),NADPH oxidase 1(nox1),glutathione peroxidases 1(gpx1),atrial natriuretic factor(anf),tumor necrosis factorα(tnfα),interleukin 6(il6),and transforming growth factorβ1b(tgfβ1b)in cardiac tissue.Results:Compared with the control group,the ventricular area and ventricular area/body weight in the model group significantly increased(P<0.05);the mRNA expression level of ptgs2,alox5a,nox1,anf,tnfα,and il6 significantly increased(P<0.05),while the mRNA expression level of gpx1 significantly decreased(P<0.05);pathology showed myocardial hypertrophy,adhesion,fibrosis,disordered arrangement of myocardial tissue,and increased apoptotic cells.Compared with the model group,the ventricular area

关 键 词：心脏毒性 阿霉素 活心丸 斑马鱼 铁死亡 炎症 

分 类 号：R285.5[医药卫生—中药学]