复合因素诱导射血分数保留心力衰竭大鼠模型的建立及评价
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1. 中国中医科学院西苑医院心血管内科,北京 100091;2. 国家中医心血管病临床医学研究中心,北京 100091

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Establishment and evaluation of a rat model of heart failure with a preserved ejection fraction induced by combined factors
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1. Department of Cardiovascular Internal Medicine, Xiyuan Hospital, Chinese Academy of Traditional Chinese Medicine, Beijing 100091, China; 2. National Clinical Research Center for Chinese Medicine Cardiology, Xiyuan Hospital of China Academy of Chinese Medical Science, Beijing 100091, China

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    摘要:

    目的 通过复合因素建立射血分数保留心力衰竭(HFpEF)大鼠模型,评估其特征,并探讨心肌应变参数与心肌肥厚及纤维化的相关性。 方法 8 只 WKY 大鼠和 8 只自发性高血压大鼠( SHR)作为对照组,给予普通饲料至实验结束。 32 只 SHR 大鼠平均分 SHR + S 组、SHR + F 组、SHR + SF 组及 SHR + 复合组,分别给予高盐饲料、高脂饲料、高盐-脂饲料、高盐-脂-糖饲料联合腹腔注射链脲霉素 30 周。 造模结束后,测量心重/ 体重(HW/BW)、收缩压( SBP) 及舒张压 ( DBP);行超声心动图测量左心室 ( LV) 舒张末内径 ( LVIDd)、 LV 前壁厚度(LVAWd)、LV 后壁厚度(LVPWd)、LV 射血分数(LVEF)、等容舒张时间( IVRT)、LV 舒张早期二尖瓣流入峰值速度(E) / 二尖瓣环运动速度(e’);斑点追踪超声心动图测量全纵向应变(GLS)及应变率(GLSr)、全径向应变(GRS)及应变率(GRSr)、全周向应变(GCS)及应变率(GCSr);血清学检测甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL-C)、葡萄糖(GLU)及糖化血清蛋白(GSP),ELISA 法检测血清 B 型利钠肽(BNP)、血管紧张素Ⅱ(AngⅡ)及半乳糖凝集素 3(Gal-3);心肌进行苏木精-伊红(HE)及 Masson 染色观察心肌细胞及纤维化,并计算心肌细胞横截面面积(CSA)及胶原体积分数(CVF);此外,分析心肌应变参数和 CSA 及 CVF 的相关性。 结果 与对照组比较,各模型组,尤其是 SHR + 复合组的 HW/ BW、SBP、DBP、血清指标(TC、TG、LDL-C、GLU、GSP、BNP、AngⅡ及 Gal-3)和超声心动图参数(LVIDd、LVAWd、LVPWd、IVRT 及 E/ e’)显著升高;斑点追踪超声心动图参数 GLS、GLSr、GRS、GRSr、GCS 及 GCSr 的绝对值显著下降;心肌组织 HE 及 Masson 染色提示明显心肌细胞肥大及纤维化,并且 CSA 及CVF 显著增加(P< 0. 05)。 相关性分析显示,GLSr、GCS 及 GCSr 与 CSA 密切相关;GLS、GLSr 和 GCSr 与 CVF 密切相关(P< 0. 01)。 结论 本研究模拟高血压及糖脂代谢紊乱诱导的 HFpEF 大鼠模型在病因、临床表现及心肌病理改变上复刻了 HFpEF 的基本特征,可能是代谢综合征相关 HFpEF 的可靠动物模型。 此外,心肌应变参数与心肌肥厚及纤维化密切相关,可能间接反应心肌细微病变和功能障碍。

    Abstract:

    Objective To evaluate the characteristics of a rat model of heart failure with a preserved ejection fraction( HFpEF) induced by combined factors, and to investigate the correlation of myocardial strain parameters to myocardial hypertrophy and fibrosis. Methods Eight WKY rats and eight spontaneously hypertensive rats(SHR)served as control groups and were fed normal feed until the end of the experiment. Thirty-two SHR rats were equally divided into SHR + S, SHR + F, SHR + SF, and SHR + Combined groups, and fed high-salt, high-fat, high-salt-fat, or high-salt-fat-sugar feed, respectively, in combination with intraperitoneal injection of streptozotocin for 30 weeks. After modeling, the heart weight / body weight(HW/ BW)ratio, systolic blood pressure( SBP), and diastolic blood pressure(DBP) were measured. Echocardiography was performed to measure the left ventricular (LV)end-diastolic internal diameter(LVIDd), LV anterior wall thickness(LVAWd), LV posterior wall thickness (LVPWd), LV ejection fraction( LVEF), isovolumetric diastolic time(IVRT), and peak early diastolic passive filling velocity( E) / early diastolic mitral annular velocity( e’). Speckle tracking echocardiography was conducted to determine the global longitudinal strain(GLS) and strain rate(GLSr), global radial strain(GRS) and strain rate(GRSr), as well as the global circumferential strain(GCS) and strain rate (GCSr).Serum was collected and analyzed for triglycerides(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDLC), glucose(GLU), and glycated serum protein (GSP). ELISA were used to measure serum B-type brain natriuretic peptide(BNP), angiotensin Ⅱ(AngⅡ), and galectin-3(Gal-3). Myocardial tissue was subjected to HE and Masson staining for cardiomyocytes and myocardial fibrosis, and the cardiomyocyte cross-sectional area (CSA)and collagen volume fraction( CVF) were calculated. Additionally, the correlation of myocardial strain parameters to CSA and CVF was analyzed. Results Compared with the control group, in model groups, especially the SHR + combined group, HW/ BW,SBP, DBP, serum indexes(TC, TG, LDL-C, GLU, GSP, BNP, AngⅡ, and Gal-3) and echocardiographic parameters (LVIDd, LVAWd, LVPWd, IVRT, and E/ e’) were significantly up-regulated. Absolute values of speckle-tracking echocardiographic parameters (GLS, GLSr, GRS, GRSr, GCS, and GCSr)were decreased considerably. HE and Masson staining of myocardial tissues suggested marked cardiomyocyte hypertrophy and fibrosis, and significant increases were observed in CSA and CVF(P< 0. 05). Correlation analysis showed that GLSr, GCS, and GCSr were strongly linked to CSA, and GLS, GLSr, and GCSr were strongly linked to CVF(P< 0. 01). Conclusions A rat model of HFpEF induced by hypertension and dysregulation of glucolipid metabolism replicated the basic characteristics of HFpEF in terms of etiology, clinical features, and myocardial pathological changes, and might be a reliable animal model of metabolic syndrome-related HFpEF. Moreover, myocardial strain indices were closely related to myocardial hypertrophy and fibrosis and might indirectly reflect subtle myocardial lesions and dysfunction.

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石玉姣,杨晨光,乔文博,刘永成,刘思雨,董国菊.复合因素诱导射血分数保留心力衰竭大鼠模型的建立及评价[J].中国实验动物学报,2024,32(3):275~285.

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  • 收稿日期:2023-07-23
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  • 在线发布日期: 2024-06-06
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