摘要
目的观察冠心宁片对慢性心力衰竭(CHF)小鼠的治疗作用,并从心肌细胞凋亡和相关细胞信号通路探讨其作用途径,为冠心宁片的临床应用提供实验依据。方法利用主动脉弓缩窄术建立小鼠CHF模型,筛选造模成功的小鼠分为模型对照组、冠心宁片低剂量组(600mg/kg)、冠心宁片高剂量组(1200mg/kg)和卡托普利阳性对照组(12.5mg/kg),并设假手术组作为对照。实验期间观察动物存活率,给药3、6、9周时进行超声成像,检测小鼠射血分数(EF),给药结束后,处死小鼠取心脏组织,TUNEL法观察心肌组织细胞凋亡情况,RT-PCR法检测BAX和BCL2基因mRNA的表达,全自动蛋白分析仪检测PI3K、p-AKT和BAX/BCL2的蛋白表达水平。结果模型对照组小鼠生存率为50%,冠心宁片低剂量组存活率为60%,冠心宁片高剂量组存活率70%。与假手术组比较,模型对照组小鼠EF值在给药3、6、9周时均显著下降,给与冠心宁片后,具有不同程度的提高;病理观察可见,模型对照组小鼠心肌细胞凋亡较多,冠心宁片能有效降低心肌细胞凋亡水平。模型对照组小鼠心肌组织中BAX基因mRNA相对表达量显著升高,BCL2下降,与蛋白表达相一致,PI3K和p-AKT水平也显著上升;与模型对照组相比,冠心宁高剂量组心肌组织中PI3K、p-AKT的蛋白表达和BAX的mRNA表达水平下降。结论冠心宁片能提高CHF小鼠的存活率,增强CHF小鼠的心脏功能,其作用机制可能是通过抑制PI3K/AKT/BAX信号通路的活化来减少心肌细胞的凋亡,从而改善CHF小鼠的心力衰竭症状。
Objective To determine the effects of guanxinning (GXN),a traditional Chinese medicine, in mice with chronic heart failure (CHF) and to explore the mechanisms involved, with particular reference to cardiomyocyte apoptosis and related signaling pathways, to provide experimental evidence for the clinical use of GXN. Methods A mouse CHF model was established by surgical aortic arch constriction and mice with successfully-induced CHF were randomly allocated to a model control group, a low-dose (600 mg/kg) GXN group, a high-dose (1,200 mg/kg) group, and a positive control group (administered 12-5 mg/kg captopril). In addition, sham-operated mice were used as a control group. The survival rate was monitored during the experiment. Ultrasonography was performed after 3, 6, and 9 weeks of the study, when the ejection fraction (EF) of the mice was measured. At the end of the study, the mice were sacrificed and their hearts collected. Myocardial apoptosis was quantified using the TUNEL staining, the mRNA expression of BAX and BCL2 were quantified using RT-PCR, and the protein expression levels of PI3K, p-AKT, and BAX/BCL2 were measured using a fully-automated protein analyzer. Results The survival rate was 50% in the model control group, 60% in the low-dose GXN group, and 70% in the high-dose GXN group. Compared with the sham surgery group, the EF in the model group was significantly lower after 3, 6, and 9 weeks, while GXN administration induced to varying degrees of improvement. There was more myocardial apoptosis in the model than in the sham group, and GXN reduced this. The myocardial mRNA expression of BAX in the model group was significantly higher, and that of BCL2 was lower, which was consistent with the protein expression levels, and the PI3K and p-AKT protein levels were also significantly higher. Compared with the model group, the mRNA expression of BAX and the protein expression levels of PI3K and p-AKT were lower in the myocardium of high-dose GXN-treated mice. Conclusions GXN improves the survival rate and cardiac function of mice with CHF. The mechanism may be involved with the upregulation of the PI3K/AKT/BAX signaling pathway, which reduces cardiomyocyte apoptosis, thereby ameliorating the signs of heart failure in CHF mice.
作者
齐月寒
马全鑫
徐松涛
郁晨
陈姣姣
方明笋
陈民利
QI Yuehan;MA Quanxin;XU Songtao;YU Chen;CHEN Jiaojiao;FANG Mingsun;CHEN Minli(Academy of Chinese Medicine/Institute of Comparative Medicine,Zhejiang Chinese Medical University,Hangzhou 310053,China)
出处
《中国比较医学杂志》
CAS
北大核心
2019年第10期33-39,共7页
Chinese Journal of Comparative Medicine
基金
浙江省中医药管理局重点项目(2015ZZ009)
浙江中医药大学科研基金(2017ZY19)
关键词
冠心宁片
慢性心力衰竭
心肌细胞凋亡
小鼠
guanxinning
chronic heart failure
myocardial cell apoptosis
mouse
