小鼠射血分数保留心力衰竭模型的建立

doi:10.3969/j.issn.1674-8115.2021.05.002

摘要/Abstract

摘要：

目的·探索小鼠射血分数保留心力衰竭（heart failure with preserved ejection fraction，HFpEF）模型的建立。方法·将8周龄的SPF级雄性和雌性C57BL/6J小鼠各16只，分别随机分成对照组和模型组，每组8只。模型组予以高脂饲料（含60%脂肪）以及含0.5 g/L N^ω-硝基-L-精氨酸甲酯盐酸盐（N^ω-nitro-L-arginine methyl ester hydrochloride，L-NAME）的饮水，对照组给予普通饲料以及饮水喂养。造模后每2周通过小动物心脏超声获取左心室二尖瓣舒张早期血流速度峰值与晚期血流速度峰值的比值（E/A比值）、二尖瓣舒张早期血流速度峰值与二尖瓣环舒张早期运动速度峰值的比值（E/E'比值）用以评估小鼠左心室舒张功能。造模后第16周，通过Masson染色和苏木精-伊红染色观察心肌细胞结构和纤维化情况。全自动生化分析仪检测小鼠血清三酰甘油（triacylglycerol，TAG）、总胆固醇（total cholesterol，TC）、高密度脂蛋白胆固醇（high-density lipoprotein cholesterol，HDL-Ch）和低密度脂蛋白胆固醇（low-density lipoprotein cholesterol，LDL-Ch）的水平。结果·雄性小鼠予以高脂饮食和L-NAME诱导后第8周出现HFpEF，雄性模型组和对照组的左室射血分数（left ventricular ejection fraction，LVEF）差异无统计学意义；雄性模型组E/A比值及E/E'比值较对照组显著升高（均P=0.000）。雌性小鼠在造模后第8周也出现了明显的HFpEF，雌性模型组与对照组的LVEF差异无统计学意义；雌性模型组E/A比值和E/E'比值较同性别对照组明显升高（P=0.000，P=0.001）。造模后第16周，雄性和雌性模型组LVEF仍与同性别对照组保持一致。雄性和雌性模型组小鼠的收缩压和舒张压均较相应性别对照组明显升高，且模型组出现左心室舒张功能明显降低，并表现为心肌细胞肥大、心肌间质纤维化和左心室重构。雄性和雌性模型组小鼠血清TAG、TC、HDL-Ch和LDL-Ch水平均较相应性别对照组明显增高。结论·应用高脂饲养联合L-NAME诱导可在雄性和雌性小鼠中建立稳定的HFpEF模型。

关键词: 心力衰竭, 射血分数保留, 动物模型

Abstract:

Objective·To establish a novel mice model of heart failure with preserved ejection fraction (HFpEF).

Methods·Sixteen 8-week-old SPF grade male and female C57BL/6J mice each were randomly divided into control group or model group (n=8 per group). The model group was given high-fat diet containing 60% fat and drinking water containing 0.5 g/L N^ω-nitro-L-arginine methyl ester hydrochloride (L-NAME). The control group was given routine feed and drinking water. The ratio of peak early mitral velocity to peak late mitral velocity (E/A ratio) and the ratio of peak early mitral velocity to peak early diastolic mitral annular velocity (E/E' ratio) were detected by echocardiography every two weeks to evaluate left ventricular diastolic function. At the 16th week after modeling, the myocardial hypertrophy and fibrosis were examined by Masson staining and hematoxylin-eosin staining. The levels of serum triacylglycerol (TAG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-Ch) and low-density lipoprotein cholesterol (LDL-Ch) were detected by automatic biochemical analyzer.

Results·HFpEF appeared in the male mice at 8 weeks after high-fat diet and L-NAME administration, with a significant increase in E/A ratio and E/E′ ratio in comparison to the male control group (both P=0.000). There was no significant difference in the left ventricular ejection fraction (LVEF) between male model group and male control group. On the other hand, the female mice fed with high-fat diet and drinking water containing L-NAME also displayed obvious HFpEF after 8 weeks. E/A ratio and E/E' ratio in female model group were significantly higher than those in the female control group (P=0.000, P=0.001). There was no significant difference in LVEF between female model group and female control group. At the 16th week after modeling, both male and female mice still displayed the characteristics of HFpEF. Systolic blood pressure and diastolic blood pressure of male and female model group were significantly higher than those of respective control groups. Male or female mice with HFpEF showed obvious myocardial hypertrophy and fibrosis along with left ventricular remodeling. The serum levels of TAG, TC, HDL-Ch and LDL-Ch in the model groups were significantly higher than those in the respective control groups.

Conclusion·High-fat feeding combined with L-NAME administration can induce HFpEF in both male and female mice.

Key words: heart failure, preserved ejection fraction, animal model

中图分类号:

R541.6

陈骁楠, 张俊峰, 王长谦, 张绘莉. 小鼠射血分数保留心力衰竭模型的建立[J]. 上海交通大学学报(医学版), 2021, 41(5): 565-570.

Xiao-nan CHEN, Jun-feng ZHANG, Chang-qian WANG, Hui-li ZHANG. Establishment of a novel mice model of heart failure with preserved ejection fraction[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(5): 565-570.

图/表 7

表1 HFpEF模型小鼠体质量的变化情况

Tab 1 Changes in body weight in the HFpEF mice

Body weight	Male			Female
Body weight	Control group （n=8）	HFpEF group （n=8）	P value	Control group （n=8）	HFpEF group （n=8）	P value
At the beginning of modeling/g	23.53±1.17	23.38±1.28	0.869	20.43±1.37	20.43±0.53	0.999
4th week/g	27.43±1.01	33.15±1.58	0.001	21.43±0.99	21.65±0.53	0.704
8th week/g	27.33±1.68	40.70±7.07	0.001	20.98±0.41	23.30±1.45	0.022
12th week/g	29.03±0.95	46.45±4.60	0.000	21.30±0.47	25.15±1.91	0.008
16th week/g	31.63±1.04	47.00±3.11	0.000	20.45±0.26	26.78±2.55	0.003

表2 HFpEF模型小鼠造模后第16周的血脂指标情况

Tab 2 Changes in blood lipid profiles in the HFpEF mice at the 16th week after modeling

Item	Male			Female			P value^①
Item	Control group （n=8）	HFpEF group （n=8）	P value	Control group （n=8）	HFpEF group （n=8）	P value	P value^①
TAG/（mmol·L^-1）	1.19±0.25	1.50±0.73	0.039	0.90±0.24	1.56±0.56	0.022	0.384
TC/（mmol·L^-1）	2.65±0.22	3.98±0.61	0.003	2.40±0.46	3.89±0.22	0.000	0.012
HDL-Ch/（mmol·L^-1）	1.43±0.11	1.89±0.36	0.032	0.84±0.18	1.37±0.08	0.001	0.035
LDL-Ch/（mmol·L^-1）	0.15±0.01	0.33±0.03	0.000	0.16±0.05	0.27±0.04	0.008	0.084

表3 HFpEF模型小鼠造模后第16周的血压情况

Tab 3 Changes in blood pressure in the HFpEF mice at the 16th week after modeling

Control group

（n=8）

HFpEF group

（n=8）

P value

Control group

（n=8）

HFpEF group

（n=8）

表4 HFpEF模型小鼠造模后第16周的心脏质量情况

Tab 4 Changes in heart weight in the HFpEF mice at the 16th week after modeling

Item

Male

Female

Control group

（n=8）

HFpEF group

（n=8）

P value

Control group

（n=8）

HFpEF group

（n=8）

Ratio of heart weight to body weight/%

图1 HFpEF模型小鼠LVEF和左心室舒张功能的超声心动图改变Note: A. LVEF. B. E/A ratio. C. E/E' ratio. D. GLS. ①P=0.000, compared with the male control group; ②P=0.000, ③P=0.001, ④P=0.002, compared with the female control group.

Fig 1 Echocardiography changes in LVEF and left ventricular diastolic function in the HFpEF mice

图2 HFpEF小鼠造模后第16周的左心室重构情况Note: A. LVEDd. B. LVAWd. C. LVPWd. ①P=0.002, compared with the female control group; ②P=0.000, compared with the male control group.

Fig 2 Left ventricular remodeling in the HFpEF mice at 16 weeks after modeling

图3 HFpEF小鼠造模后第16周后心肌组织的病理组织学变化Note: A. Masson staining in heart sections of mice. B. Hematoxylin-eosin staining in heart sections of mice. C. Quantitative analysis of fibrosis. D. Quantitative analysis of inflammatory cells infiltration. ①P=0.007, ②P=0.013, ③P=0.012, ④P=0.046, compared with the respective control group.

Fig 3 Pathological characteristics in cardiomyocytes of the HFpEF mice at 16 weeks after modeling

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