文章摘要
张云燕,丁君灿,王天宇,等.芪附强心方改善心肾阳虚型心衰小鼠心肌损伤的机制研究[J].浙江中医药大学学报,2024,48(3):264-272.
芪附强心方改善心肾阳虚型心衰小鼠心肌损伤的机制研究
Mechanism of Qifu Qiangxin Decoction Relieving Myocardial Damage in Heart Failure Mice with Heart-Kidney Yang Deficiency Syndrome
DOI:10.16466/j.issn1005-5509.2024.03.003
中文关键词: 芪附强心方  心功能  心肌损伤  心肌细胞凋亡  自噬  AMPK/mTOR信号通路
英文关键词: Qifu Qiangxin Decoction  cardiac function  myocardial injury  cardiomyocyte apoptosis  autophagy  AMPK/mTOR signaling pathway
基金项目:浙江省中医药管理局重点研究项目(2022ZZ081);2021年浙江中医药大学附属医院科研专项(2021FSYYZZ12);衢州市科技攻关项目第二批中医药专项(2022K114)
作者单位
张云燕 浙江中医药大学第二临床医学院 杭州 310053 
丁君灿 浙江中医药大学第二临床医学院 杭州 310053 
王天宇 浙江中医药大学第二临床医学院 杭州 310053 
邱渭 浙江中医药大学第二临床医学院 杭州 310053 
华军益 浙江中医药大学第二临床医学院 杭州 310053
浙江中医药大学附属第二医院 
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中文摘要:
      [目的] 探究芪附强心方缓解心肾阳虚型心衰(heart failure,HF)小鼠心肌损伤的作用机制。[方法] 30只C57BL/6小鼠随机分为假手术(Sham)组、芪附强心方低剂量(HF+QL)组、芪附强心方高剂量(HF+QH)组、西药[HF+血管紧张素转化酶抑制剂(angiotensin converting enzyme inhibitors,ACEI)]组和模型(HF)组,每组6只。Sham组小鼠麻醉后切开皮肤,暴露心脏,不进行左冠状动脉前降支结扎术,然后缝合;其余组通过结扎左冠状动脉前降支配合冷水力竭式游泳,制备心梗后心肾阳虚型HF小鼠模型。治疗15 d后,记录小鼠状态;以超声心动图检测左心室舒张末期容积(left ventricular end-diastolic volume,LVEDV)、左心室收缩末期容积(left ventricular end-systolic volume,LVESV)、射血分数(ejection fraction,EF)和左室短轴缩短率(left ventricular fractional shortening,LVFS)评估心功能; 苏木精-伊红(hematoxylin-eosin,HE)染色评估心肌组织的形态学改变;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测小鼠血清脑钠肽(B-syndrome natriuretic peptide,BNP)水平;脱氧核苷酸末端转移酶介导的dUTP 缺口末端标记(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling,TUNEL)检测小鼠心肌细胞凋亡情况;免疫印迹法检测小鼠心肌组织中凋亡相关蛋白、自噬相关蛋白和腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白(adenosine monophosphate-activated protein kinase/mammalian target of rapamycin,AMPK/mTOR)信号通路相关蛋白的表达水平。[结果] 与Sham组比较,HF组小鼠活动量、饮食饮水显著减少,精神状态变差,便溏,毛发枯燥,EF值和LVFS值均显著降低,LVEDV和LVESV显著升高(P<0.01);HE染色可见广泛坏死的心肌组织;ELISA提示血清BNP水平显著升高(P<0.01)。与HF组比较,给药各组小鼠活动量、饮食饮水显著增加,精神状态变好,便溏和毛发枯燥情况显著改善,EF值和LVFS值均显著上升,LVEDV和LVESV显著降低(P<0.01),且HF+QH组效果优于HF+QL组;HE染色可见给药组小鼠心肌组织的病理情况改善;ELISA结果显示,各给药组小鼠血清BNP水平也显著降低(P<0.01)。TUNEL染色和免疫印迹结果显示,与Sham组比较,HF组小鼠心肌细胞凋亡水平显著增加(P<0.05);与HF组比较,给药各组小鼠心肌细胞凋亡水平显著降低(P<0.05)。免疫印迹结果显示,芪附强心方可以显著增强自噬水平,调控AMPK/mTOR信号通路。[结论] 芪附强心方可调控AMPK/mTOR信号通路,抑制心肌细胞凋亡,诱导自噬,从而保护心肌细胞,缓解心肌损伤。
英文摘要:
      [Objective] To explore the mechanism of Qifu Qiangxin Decoction mitigating myocardial damage in heart failure(HF) mice with heart-kidney Yang deficiency syndrome. [Methods] Thirty C57BL/6 mice were randomly divided into Sham surgery group(Sham group), HF model(HF) group, low-dose Qifu Qiangxin Decoction(HF+QL) group, high-dose Qifu Qiangxin Decoction(HF+QH) group and western medicine[HF+angiotensin converting enzyme inhibitors(ACEI)] group, six in each group. In Sham group, the skin was cut open after anesthesia, the heart was exposed, the left anterior descending coronary artery was not in ligation, and then sutured. The rest were used to establish a mouse model of HF with heart-kidney Yang deficiency syndrome after myocardial infarction(MI) by ligating the left anterior descending coronary artery and swimming in cold water, then treated for 15 days. After treatment, the state of the mice was recorded, left ventricular end-diastolic volume(LVEDV), left ventricular end-systolic volume(LVESV), ejection fraction(EF) and left ventricular fractional shortening(LVFS) were measured by echocardiography to evaluate cardiac function; hematoxylin-eosin(HE) staining was used to evaluate the morphological of myocardial tissue; the serum levels of B-syndrome natriuretic peptide(BNP) were measured by enzyme linked immunosorbent assay(ELISA); terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL) was used to detect cardiomyocyte apoptosis; Western blot was used to determine the expression levels of apoptosis related proteins, autophagy related proteins and adenosine monophosphate-activated protein kinase/mammalian target of rapamycin(AMPK/mTOR) signaling pathway related proteins in mice myocardial tissue. [Results] Qifu Qiangxin Decoction can relieve the symptoms of HF in mice. Compared with Sham group, EF and LVFS values of mice in HF group were significantly decreased, while LVEDV and LVESV were significantly increased(P<0.01). Compared with HF group, EF and LVFS values in each group were significantly increased, while LVEDV and LVESV were significantly decreased(P<0.01), moreover, HF+QH group had a better effect than that of HF+QL group. According to HE staining, extensive necrotic myocardial tissue was observed in HF group compared with Sham group, and ELISA showed a significant increase in BNP levels(P<0.01). Compared with HF group, the pathological conditions of myocardial tissue were relieve in each group, and the level of BNP was also significantly reduced(P<0.01). TUNEL staining and Western blot results showed that the level of apoptosis in HF group was significantly increased compared with Sham group(P<0.05). Compared with HF group, the apoptosis level of the each group was significantly reduced(P<0.05). Therefore, Qifu Qiangxin Decoction could significantly reduce the level of cardiomyocyte apoptosis. Western blot detection of autophagy-related proteins and AMPK/mTOR signaling pathway related proteins showed that Qifu Qiangxin Decoction could significantly enhance autophagy level and regulate AMPK/mTOR signaling pathway in a concentration-dependent manner. [Conclusion] Qifu Qiangxin Decoction can regulate AMPK/mTOR signaling pathway, inhibit cell apoptosis and induce autophagy, thus protecting cardiomyocytes and mitigating myocardial injury.
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