心肌缺血和动脉粥样硬化斑块血管新生双靶点小鼠模型的建立
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摘要
目的探讨建立心肌缺血和动脉粥样硬化(atherosclerosis,AS)斑块内血管新生双靶点小鼠模型的有效方法。方法 C57BL/6 J小鼠10只作为对照组,Apo E-/-小鼠10只作为模型组。对照组喂普通饲料,模型组喂高脂饲料。模型组高脂饲料喂养8周后,皮下注射异丙肾上腺素100 mg/kg,连续2 d,对照组皮下注射等量生理盐水。4周后,检测血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(lowdensity lipoprotein cholesterol,LDL-C)和高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)水平; HE染色观察主动脉和心肌不同区域的病理形态学变化; CD31免疫组化染色检测主动脉和心肌组织不同部位的新生血管密度。结果模型组血TC、LDL-C水平显著高于对照组(P<0. 05),而HDL-C显著低于对照组(P<0. 05); HE染色显示模型组主动脉形成了典型的AS病理学改变;皮下注射异丙肾上腺素4周后,模型组心肌组织HE染色可见典型的心肌梗死病理改变; CD31免疫组化染色结果显示,模型组主动脉新生血管密度显著高于对照组(P<0. 05),模型组心肌缺血区的新生血管表达最丰富,显著高于心肌梗死区和正常心肌组织区(P <0. 05)。结论Apo E-/-小鼠在高脂饮食下,通过皮下注射异丙肾上腺素造成心肌梗死,能够成功建立心肌缺血和AS斑块血管新生的双靶点小鼠模型。
Objective To explore an effective method for establishing a dual-target mouse model of angiogenesis in myocardial ischemic and atherosclerotic plaque. Methods Ten C57 BL/6 J mice were fed basic forage as a control group,while ten Apo E-/-mice were fed high-fat forage as a model group. After 8 weeks, the model group was subcutaneously injected with isoproterenol at 100 mg/kg once a day for 2 days,while the control group was subcutaneously injected with the same amount of physiological saline. Another 4 weeks later,the serum levels of TC,TG,LDL-C,andHDL-C were measured. HE staining was used to estimate the pathomorphological changes in aortic tissues and different regions of myocardium. Immunohistochemical staining was used to measure the density of new vessels in aortic plaques and different regions of the myocardium that were marked by CD31. Results The levels of serum TC and LDL-C in the model group were significantly higher than those in the control group( P < 0. 05),while HDL-C was significantly lower than that in the control group( P < 0. 05). HE staining revealed pathological changes typical of AS in the aorta of the model group.After 4 weeks of the subcutaneous injection of isoproterenol,pathological changes typical of myocardial infarction were observed in HE staining of myocardial tissue in the model group. CD31 immunohistochemical staining showed that the density of new vessels in aorta of the model group was significantly higher than that in the control group( P < 0. 05). The expression of neovascularization in the myocardial ischemia zone was the highest in the model group,which was significantly higher than that in the myocardial infarction zone and normal myocardium zone( P < 0. 05). Conclusions ApoE-/-mice with myocardial infarction induced by subcutaneous injection of isoproterenol under a diet of high-fat forage can be successfully used to establish a dual-target mouse model of angiogenesis in myocardial ischemia and atherosclerotic plaque.
Objective To explore an effective method for establishing a dual-target mouse model of angiogenesis in myocardial ischemic and atherosclerotic plaque. Methods Ten C57 BL/6 J mice were fed basic forage as a control group,while ten Apo E-/-mice were fed high-fat forage as a model group. After 8 weeks, the model group was subcutaneously injected with isoproterenol at 100 mg/kg once a day for 2 days,while the control group was subcutaneously injected with the same amount of physiological saline. Another 4 weeks later,the serum levels of TC,TG,LDL-C,andHDL-C were measured. HE staining was used to estimate the pathomorphological changes in aortic tissues and different regions of myocardium. Immunohistochemical staining was used to measure the density of new vessels in aortic plaques and different regions of the myocardium that were marked by CD31. Results The levels of serum TC and LDL-C in the model group were significantly higher than those in the control group( P < 0. 05),while HDL-C was significantly lower than that in the control group( P < 0. 05). HE staining revealed pathological changes typical of AS in the aorta of the model group.After 4 weeks of the subcutaneous injection of isoproterenol,pathological changes typical of myocardial infarction were observed in HE staining of myocardial tissue in the model group. CD31 immunohistochemical staining showed that the density of new vessels in aorta of the model group was significantly higher than that in the control group( P < 0. 05). The expression of neovascularization in the myocardial ischemia zone was the highest in the model group,which was significantly higher than that in the myocardial infarction zone and normal myocardium zone( P < 0. 05). Conclusions ApoE-/-mice with myocardial infarction induced by subcutaneous injection of isoproterenol under a diet of high-fat forage can be successfully used to establish a dual-target mouse model of angiogenesis in myocardial ischemia and atherosclerotic plaque.
引文
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[4] Guo D,Murdoch CE,Liu T,et al. Therapeutic angiogenesis of chinese herbal medicines in ischemic heart disease:A Review[J]. Front Pharmacol,2018,9:428.
[5] Wu W,Li X,Zuo G,et al. The role of angiogenesis in coronary artery disease:a double-edged sword:intraplaque angiogenesis in physiopathology and therapeutic angiogenesis for Treatment[J].Curr Pharm Des,2018,24(4):451-464.
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[7] Zukowska Z. Atherosclerosis and angiogenesis:what do nerves have to do with it?[J] Pharmacol Rep,2005,57 Suppl:229-234.
[8] Emini Veseli B,Perrotta P,De Meyer GRA,et al. Animal models of atherosclerosis[J]. Eur J Pharmacol,2017,816:3-13.
[9]兰琴,王竹林,曹浩,等.改良左旋支结扎法微创小型猪急性心肌梗死模型建立与评估[J].中国实验动物学报,2017,25(4):414-419.
[10]曹珏,李贻奎,陈孟倩,等.大鼠急性心肌梗死模型制备方法研究进展[J].中国比较医学杂志,2017,27(10):96-100.
[11] Lobo Filho HG,Ferreira NL,Sousa RB,et al. Experimental model of myocardial infarction induced by isoproterenol in rats[J]. Rev Bras Cir Cardiovasc,2011,26(3):469-476.
[2] Perrotta P, Emini Veseli B, Van der Veken B, et al.Pharmacological strategies to inhibit intra-plaque angiogenesis in atherosclerosis[J]. Vascul Pharmacol,2018,pii:S1537-1891(18)30113-30117.
[3]刘灿,海宝,张稳,等.骨内单次注射小剂量辛伐他汀对大鼠心梗后血管新生的影响[J].中国实验动物学报,2017,25(5):506-512.
[4] Guo D,Murdoch CE,Liu T,et al. Therapeutic angiogenesis of chinese herbal medicines in ischemic heart disease:A Review[J]. Front Pharmacol,2018,9:428.
[5] Wu W,Li X,Zuo G,et al. The role of angiogenesis in coronary artery disease:a double-edged sword:intraplaque angiogenesis in physiopathology and therapeutic angiogenesis for Treatment[J].Curr Pharm Des,2018,24(4):451-464.
[6] Yuan R, Shi WL, Xin QQ, et al. Holistic regulation of angiogenesis with Chinese herbal medicines as a new option for coronary artery disease[J]. Evid Based Complement Alternat Med,2018,2018:3725962.
[7] Zukowska Z. Atherosclerosis and angiogenesis:what do nerves have to do with it?[J] Pharmacol Rep,2005,57 Suppl:229-234.
[8] Emini Veseli B,Perrotta P,De Meyer GRA,et al. Animal models of atherosclerosis[J]. Eur J Pharmacol,2017,816:3-13.
[9]兰琴,王竹林,曹浩,等.改良左旋支结扎法微创小型猪急性心肌梗死模型建立与评估[J].中国实验动物学报,2017,25(4):414-419.
[10]曹珏,李贻奎,陈孟倩,等.大鼠急性心肌梗死模型制备方法研究进展[J].中国比较医学杂志,2017,27(10):96-100.
[11] Lobo Filho HG,Ferreira NL,Sousa RB,et al. Experimental model of myocardial infarction induced by isoproterenol in rats[J]. Rev Bras Cir Cardiovasc,2011,26(3):469-476.