丹参酮ⅡA通过半乳糖凝集素-3抑制心肌重塑的机制研究  

Mechanism of tanshinoneⅡA inhibiting myocardial remodeling by Galectin-3

作　　者：张美齐 程抗 余利美 吴微华 王亚坤[1] 陈雲 Zhang Meiqi;Cheng Kang;Yu Limei;Wu Weihua;Wang Yakun;Chen Yun(Hangzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University,Hangzhou 310021,Zhejiang,China)

机构地区：[1]浙江中医药大学附属杭州市中医院丁桥院区重症医学科,杭州310021

出　　处：《中华危重病急救医学》2022年第6期640-645,共6页Chinese Critical Care Medicine

基　　金：浙江省中医药管理局重点项目(2018ZZ001)。

摘　　要：目的分析丹参酮ⅡA对缺血/再灌注(I/R)所致心力衰竭(心衰)模型大鼠心肌重构的影响。方法①体内实验:将30只健康雄性SD大鼠按随机数字表法分为假手术组、心衰模型组和丹参酮ⅡA组,每组10只。采用结扎大鼠左冠状动脉(冠脉)待心电监护出现明显ST段抬高30 min后放松结扎线进行再灌注2 h来制备I/R后心衰模型;假手术组于同期开胸,仅丝线绕过左冠脉而不结扎。丹参酮ⅡA组术后3 d开始腹腔注射丹参酮ⅡA 10 mg/kg,连续用药9周;其他两组于同期腹腔注射等量生理盐水。干预9周后观察各组大鼠的行为学表现,检测血流动力学相关指标。处死大鼠取心脏组织,Masson染色后光镜下观察心肌组织纤维化程度;采用酶联免疫吸附试验(ELISA)检测半乳糖凝集素-3(Galectin-3)含量;采用荧光定量反转录-聚合酶链反应(qRT-PCR)检测Ⅰ型胶原、Ⅲ型胶原、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制剂-1(TIMP-1)的mRNA表达;采用蛋白质免疫印迹试验(Western blotting)检测MMP-2和TIMP-1的蛋白表达。②体外实验:提取并分离大鼠的原代心肌成纤维细胞,分为空白对照组、血管紧张素Ⅱ组(给予7~10 mmol/L血管紧张素Ⅱ)和血管紧张素Ⅱ+丹参酮ⅡA组(同时给予7~10 mmol/L血管紧张素Ⅱ+5~10 mmol/L丹参酮ⅡA)。分别于培养24 h和48 h时,采用四甲基偶氮唑蓝(MTT)检测细胞吸光度(A)值并计算细胞增殖率;采用qRT-PCR法检测型胶原、型胶原、MMP-2和TIMP-1的mRNA表达;采用ELISA法检测Galectin-3含量。结果①体内实验:大鼠活动状态、毛发顺帖程度及进食量由好到差依次为假手术组、丹参酮ⅡA组和心衰模型组。与假手术组相比,心衰模型组大鼠心率(HR)明显下降、心功能明显受损,Ⅰ型胶原、Ⅲ型胶原、TIMP-1的mRNA和蛋白表达及Galectin-3含量均明显升高,MMP-2的mRNA和蛋白表达均明显降低。与心衰模型组相比,丹参酮ⅡA组大鼠HR明显升高�Objective To explore the effect of tanshinoneⅡA on myocardial remodeling in ischemia/reperfusion(I/R)-induced heart failure of rodent model.Methods①In vivo,30 SD rats were randomly divided into sham operation,heart failure and tanshinoneⅡA treatment group,with 10 rats in each group.The I/R model was established by ligating the left coronary artery until ST segment elevation for 30 minutes,then the ligation was removed for 2 hours as reperfusion.In the sham operation group,the rat chest was opened without artery ligation.Three days after model establishment,tanshinoneⅡA(10 mg/kg)were given intraperitoneal injected in tanshinoneⅡA group for 9 weeks.In the other two groups,normal saline was administrated in the same way.The behavioral manifestations of the rats in each group were observed;hemodynamic indexes were evaluated;Masson staining was performed to observe the degree of myocardial fibrosis;enzyme linked immunosorbent assay(ELISA)was used to detect the content of Galectin-3 in myocardial tissue;quantitative reverse transcription-polymerase chain reaction(qRT-PCR)was used to detect the expressions of collagenⅢ,collagenⅠ,matrix metalloproteinase 2(MMP-2)and tissue inhibitor of metalloproteinase(TIMP-1).②In vitro,rats primary cardiac fibroblasts were extracted and isolated,and divided into blank control group,angiotensinⅡgroup(7-10 mmol/L angiotensinⅡ)and angiotensinⅡ+tanshinoneⅡA group(7-10 mmol/L angiotensinⅡ+5-10 mmol/L tanshinoneⅡA).At 24 hours and 48 hours of culture,the cell proliferation in each group was detected by methyl thiazolyl tetrazolium(MTT);the expressions of collagenⅢ,collagenⅠ,MMP-2 and TIMP-1 were detected by qRT-PCR;the content of Galectin-3 in cardiac fibroblasts was detected by ELSIA.Results①In vivo,the rats'activity status,hair conformity and food intake were ranked from good to bad in order of sham operation group,tanshinoneⅡA group and heart failure model group.Compared with the sham-operated group,the heart rate(HR)of the rats in the heart failure mod

关 键 词：丹参酮ⅡA 半乳糖凝集素-3 心肌重构 心肌纤维化 

分 类 号：R285.5[医药卫生—中药学]