不同给药方式建立异丙肾上腺素诱导小鼠心肌纤维化模型的评价

Evaluation of Isoproterenol-induced Myocardial Fibrosis Model in Mice by Different Administration Methods

目的评价2种不同的给药方式建立异丙肾上腺素(isoproterenol,ISO)诱导小鼠心肌纤维化模型的效果。方法选取60只8~10周龄的雄性C57BL/6J小鼠,随机分为对照组、模型组A和模型组B,每组各20只。对照组小鼠给予腹腔注射0.9%氯化钠溶液,模型组A小鼠腹腔注射ISO[5mg/(kg·d)],模型组B小鼠皮下多点注射ISO[5mg/(kg·d)],每天1次,连续给药21天。在末次给药24h后采用小动物超声仪检测小鼠心功能,计算左心室重量(left ventricular mass,LV mass)、左心室射血分数(left ventricular ejection fraction,LVEF)和左心室短轴缩短率(left ventricular fractional shortening,LVFS),并计算不同时间点的小鼠死亡率;小鼠安乐死后取材计算心脏重量与体质量比率(HW/BW)、心脏重量与胫骨长度比率(HW/TL)和肺脏重量与体质量比率(LW/BW);通过Masson染色观察心肌纤维化的程度;Western blot法检测小鼠心脏组织胶原蛋白表达水平。结果与对照组比较,模型组A和模型组B小鼠均出现心脏重量增加,心室腔变大,心功能下降,心肌纤维增生,心肌胶原蛋白表达增加,表明心肌纤维化模型构建成功。与模型组B比较,模型组A小鼠的生存率、EF和FS值较低(P<0.05),LW/BW值较高(P<0.05),表明模型组A小鼠出现心力衰竭。心肌胶原蛋白定量和Masson染色结果表明,模型组A和模型组B小鼠纤维化相关指标比较,差异无统计学意义(P>0.05)。结论腹腔注射和皮下多点注射ISO均可成功建立小鼠心肌纤维化模型,但皮下多点注射方法构建小鼠心肌纤维化模型的效果更好。

Objective To evaluate the effects of establishing isoproterenol(ISO)-induced myocardial fibrosis mice model by two different administration methods.Methods Sixty male C57BL/6J mice aged 8-10 weeks were randomly divided into 3 groups,with 20mice in each group,namely control group,model group A and B.Mice in control group were given 0.9%sodium chloride solution,mice in group A was intraperitoneally injected ISO[5mg/(kg·d)],and mice in group B was subcutaneously multipoint injected ISO[5mg/(kg·d)],once a day,for 21days.24h after the last administration,cardiac function was detected by ultrasound apparatus,left ventricular mass(LV mass)mass,left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),and mortality of mice during different time were calculated.Heart weight index and lung weight index of mice were calculated after euthanasia,;the degree of myocardial fibrosis were observed by Masson staining;the expression of collagen protein in the heart tissue of mice were detected by Western blot method.Results Compared with control group,the mice in group A and group B showed heart weight increased,ventricular cavity enlarged,cardiac function decreased,myocardial fiber hyperplasia and myocardial collagen expression increased,indicating that the myocardial fibrosis model was successfully constructed.Compared with group B,survival rate,EF and FS of group A were lower(P<0.05),and LW/BW was higher(P<0.05),indicating that cardiac function of group A mice had already exhibited heart failure.Myocardial collagen quantification and Masson staining showed that there was no significant difference in fibrosis-related indexes between group A and group B(P>0.05).Conclusion Both intraperitoneal injection and subcutaneous multipoint injection of ISO can successfully establish mice myocardial fibrosis model,but subcutaneous multipoint injection method is more effective.