异丙肾上腺素诱导慢性心衰模型的方法优化

Method optimization of an isoproterenol-induced chronic heart failure model

目的目前对于慢性心力衰竭(chronic heart failure,CHF)发病机制的研究和治疗手段均具有局限性,为了更好地研究其发病机理,本文对异丙肾上腺素(isoproterenol,ISO)诱导慢性心力衰竭大鼠模型的制备方法进行了优化研究。方法将SD雄性大鼠分为4组,模型A组大鼠连续以5 mg/(kg·d)注射ISO 10 d;模型B组大鼠前3 d注射ISO 2.5 mg/(kg·d),后9 d注射5 mg/(kg·d);模型C组大鼠第1天被给予15 mg/kg的戊巴比妥钠,注射ISO 1.25 mg/kg,第2、3天以2.5 mg/(kg·d)注射,后9 d以5 mg/(kg·d)注射,从第2天开始每天分2次注射间隔30 min。对照组给予等量生理盐水。通过超声心动图、病理学检测和荧光定量PCR等方法来验证模型的建立。结果模型A、B和C组的死亡率分别为80%、60%和40%。与对照组相比,模型C组大鼠的左心室短轴缩短率和射血分数显著降低(P<0.001);心肌细胞排列紊乱,Ⅰ、Ⅲ型胶原表达显著增加(P<0.05);COL1A1、COL3A1、FN1和ACTA2 mRNA表达水平显著上调(P<0.05);血清氨基末端脑钠肽前体(N-terminal pro-B type natriuretic peptide,NT-pro BNP)与甘油三脂的水平显著升高(P<0.05)。结论注射麻醉剂使大鼠处于半清醒状态,梯度增加每日注射剂量和减少单次注射量均可降低死亡率,为建立CHF大鼠模型提供新思路与新方式。

Objective Current research and treatments for the pathogenesis of chronic heart failure(CHF)are limited.In order to better study its pathogenesis,this paper optimized the preparation method of isoproterenol(ISO)-induced chronic heart failure rat model.Methods SD male rats were divided into 4 groups,and rats in model A were injected with ISO 5 mg/(kg·d)continuously for 10 days;rats in model B were injected with ISO 2.5 mg/(kg·d)on the first 3 days and 5 mg/(kg·d)on the last 9 days;rats in model C were given 15 mg/kg of pentobarbital sodium on day 1 and ISO 1.25 mg/kg on day 2 and 3,and 2.5 mg/(kg·d)on day 2 and 3,and 5 mg/(kg·d)on the last 9 days,with 30 min interval between 2 injections per day from day 2.The control group was given equal amounts of saline.The successful establishment of the model was verified by echocardiography,pathology testing and fluorescence quantitative PCR.Results Mortality rates were 80%,60%,and 40% in model groups A,B,and C,respectively.Compared with the control group,the left ventricular short axis shortening and ejection fraction were significantly lower in model group C(P<0.001).Rats in group C had disturbed myocardial cell arrangement,significantly increased collagen type Ⅰ and Ⅲ expression(P<0.05),and significantly upregulated expression levels of fibrosis indexes COL1A1,COL3A1,FN1 and ACTA2 mRNA(P<0.05),while the levels of serum N-terminal pro-B type natriuretic peptide(NT-pro BNP)and triglycerides were significantly increased(P<0.05).Conclusions Injecting anesthetics to keep the rats in a semi-conscious state,increasing the daily injection dose and decreasing the single injection dose in a gradient can reduce the mortality rate,which can provide a new idea and a new approach for establishing a CHF rat model.