5-羟基色氨酸对阿霉素诱导的心肌细胞损伤的保护作用及其机制  

PROTECTIVE EFFECT OF 5-HYDROXYTRYPTOPHAN AGAINST DOXORUBICIN-INDUCED CARDIOMYOCYTE INJURY AND ITS MECHANISM

作　　者：张钰坤 原阳 卢琦[1,2] 邹林峰 高远真 邢东明 ZHANG Yukun;YUAN Yang;LU Qi;ZOU Linfeng;GAO Yuanzhen;XING Dongming(School of Ba-sic Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院,山东青岛266071 [2]青岛大学附属医院肿瘤研究所,青岛肿瘤研究院 [3]清华大学生命科学学院

出　　处：《精准医学杂志》2022年第3期222-227,共6页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(32000830);中国博士后科学基金资助项目(2021T140356);青岛市高层次人才团队引进计划项目(17212zhc)。

摘　　要：目的探讨5-羟基色氨酸(5-HTP)对阿霉素(Dox)诱导的心肌损伤的保护作用及其机制。方法将H9c2细胞随机分为对照组、模型组(5μmol/L Dox)、阳性药物保护组(5μmol/L Dox+20μmol/L Dexra)、5-HTP保护组(5μmol/L Dox+20μmol/L 5-HTP),经药物干预24 h后,分别测定各组细胞的细胞存活率、乳酸脱氢酶(LDH)漏出量、细胞凋亡情况、活性氧(ROS)水平、超氧化物歧化酶(SOD)活性、三磷酸腺苷(ATP)水平及线粒体膜电位变化情况;采用Western Blot方法检测各组细胞中小G蛋白Rho相关激酶1(Rock-1)、核因子E2相关因子2(Nrf-2)、沉默调节蛋白1(Sirt-1)、B淋巴细胞瘤-2(Bcl-2)蛋白的表达水平。将32只雌性C57BL/6J小鼠随机分为对照组、模型组(仅第1天腹腔注射Dox 10 mg/kg)、阳性药物保护组(模型组处理基础上再腹腔注射14.3 mg/kg Dexra连续7 d)和5-HTP保护组(模型组处理基础上再用5-HTP 30 mg/kg连续灌胃7 d),通过超声心电图检测各组小鼠射血分数(EF%)和短轴缩短率(FS%),使用HE染色评估小鼠心脏组织形态,测定小鼠血清中心肌肌钙蛋白的含量。RT-qPCR法测定对照组、模型组及5-HTP保护组小鼠心脏组织中凋亡相关基因含半胱氨酸的天冬氨酸蛋白水解酶1(Casp 1)、Casp 2、B淋巴细胞瘤-2关联X(Bax)及B淋巴细胞瘤-2基因相关启动子(Bad)的相对表达水平。结果对照组和5-HTP保护组H9c2细胞的LDH漏出量、细胞凋亡率、ROS水平、SOD活性、线粒体膜电位、ATP水平及Bcl-2、Sirt-1、Nrf-2、Rock-1蛋白表达水平与模型组相比,差异均有显著性(F=17.15~238.27,P<0.05),阳性药物保护组细胞的ROS水平、SOD活性、线粒体膜电位及Bcl-2、Nrf-2和Rock-1蛋白表达水平与5-HTP保护组相比,差异有显著性(P<0.05)。对照组和5-HTP保护组小鼠的EF%、FS%与血清中心肌肌钙蛋白水平以及心脏组织中基因Casp 1、Casp 2、Bax及Bad的相对表达水平与模型组相比,差异有显著性(F=6.91~59.69,P<0.05)。HE染色Objective To investigate the protective effect of 5-hydroxytryptophan(5-HTP)against doxorubicin(Dox)-induced myocardial injury and its mechanism.Methods H9c2 cells were randomly divided into control group,model group(5μmol/L Dox),positive drug-protected group(5μmol/L Dox+20μmol/L Dexra),and 5-HTP-protected group(5μmol/L Dox+20μmol/L 5-HTP),and related indicators were measured for each group after 24 hours of drug intervention,including cell viability,lactate dehydrogenase(LDH)leakage,cell apoptosis,reactive oxygen species(ROS)level,superoxide dismutase(SOD)activity,adenosine triphosphate(ATP)level,and change in mitochondrial membrane potential.Western blot was used to measure the protein expression levels of rho-associated protein kinase 1(Rock-1),nuclear factor erythroid 2-related factor 2(Nrf-2),silent information regulator 1(Sirt-1),and B-cell lymphoma-2(Bcl-2)in each group of cells.A total of 32 female C57BL/6J mice were randomly divided into control group,model group(intraperitoneal injection of Dox 10 mg/kg on day 1 only),positive drug protection group(intraperitoneal injection of 14.3 mg/kg Dexra for 7 consecutive days in addition to the treatment in the model group),and 5-HTP protection group(5-HTP 30 mg/kg by gavage for 7 consecutive days in addition to the treatment in the model group).Echocardiography was used to measure ejection fraction(EF%)and fractional shortening(FS%),HE staining was used to eva-luate cardiac histomorphology,and the serum level of cardiac troponin was measured.RT-qPCR was used to measure the relative mRNA expression levels of the apoptosis-related genes cysteine-aspartic acid protease 1(Casp1),cysteine-aspartic acid protease 2(Casp2),Bcl-2 related X gene(Bax),and Bcl-2-associated agonist of cell death(Bad)in the heart tissue of mice in the control group,the model group,and the 5-HTP protection group.Results In H9c2 cardiomyocytes,there were significant differences between the control/5-HTP-protected groups and the model group in LDH leakage,cell apoptosis rate,ROS level,SOD a

关 键 词：多柔比星 心脏毒性 5-羟色氨酸 氧化性应激 线粒体 能量代谢 细胞凋亡 

分 类 号：R541[医药卫生—心血管疾病]