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高脂饮食结合球囊损伤快速建立ApoE-/-大鼠动脉粥样硬化模型
作者:杨炯 张耀雷 代凤丹 韦舒杰 王强 李德 杨大春 
单位:400042 重庆 第三军医大学研究生管理大队(杨炯) 610083 成都 成都军区总医院心血管内科(杨炯、代凤丹、韦舒杰、王强、李德、杨大春) 中心实验室(张耀雷) 
关键词:基因敲除技术 球囊损伤 高脂血症 动脉粥样硬化 
分类号:R543.5
出版年,卷(期):页码:2016,41(12):982-986
摘要:
[摘要] 目的 探索利用短期高脂饮食结合球囊损伤的造模方式,快速建立载脂蛋白E基因敲除(ApoE-/-)大鼠动脉粥样硬化(AS)动物模型。方法 选择8周雄性SD大鼠(n=10)ApoE-/-大鼠(n=10),高脂饮食2周后分别检测两组血常规、肝肾功能、血脂谱及C反应蛋白(CRP)等指标。随后行大鼠左侧颈总动脉球囊损伤术,2周后采用过量水合氯醛处死大鼠,取术侧颈总动脉进行HEMasson及油红O染色,并行CD68α平滑肌蛋白(α-SMA)免疫荧光染色。结果 ApoE-/-大鼠血脂水平明显高于SD大鼠[总胆固醇(TC)18.56±2.82mmol/L vs 5.69±1.98mmol/LP<0.01;低密度脂蛋白(LDL)6.86±1.47mmol/L vs 1.92±0.76mmol/LP<0.01];与SD大鼠比较,在严重血脂紊乱状态下,ApoE-/-大鼠全身处于显著炎症状态[CRP4.66±0.57mg/L vs 0.39±0.21mg/LP<0.05;白细胞:(21.79±5.10)×109/L vs (14.82±2.41)×109/LP<0.01;中性粒细胞:(9.28±3.35)×109/L vs (2.10±0.96)×109/LP<0.01]HEMasson染色结果显示,两组均有显著增生形成,并伴随着胶原纤维的轻度沉积;油红O染色可见ApoE-/-大鼠斑块增生明显,而SD大鼠呈阴性。免疫荧光染色结果显示,ApoE-/-大鼠斑块内CD68呈显著阳性,α-SMA呈弱阳性,而SD大鼠α-SMA呈阳性,无明显CD68显色。结论 通过短期高脂饮食结合球囊损伤的造模方式,可以快速建立ApoE-/-大鼠AS模型。

[Abstract] Objective To quickly establish the atherosclerosis (AS) model in apoprotein E gene knockout (ApoE-/-) rats by short-term high-fat diet combined with balloon injury. Methods Eight-week-old male SD and ApoE-/- rats (10 each) were used in the experiments. After fed with high-fat diet for 2 weeks, the blood routine, hepatorenal function, blood lipid profile and C-reactive protein (CRP) were measured, and then all the left common carotid arteries were processed by balloon injury. After 2 weeks, rats euthanasia were carried out by excessive chloral hydrate, and then the operation side common carotid arteries were stained with HE, Masson trichrome and oil red O. While the common carotid arteries accepted CD68, α-SMA immunofluorescent staining. Results The blood lipid level was significantly higher in ApoE-/- rats than in SD rats [total cholestrol (TC): 18.56±2.82mmol/L vs 5.69±1.98mmol/L, P<0.01; low density lipoprotein (LDL): 6.86±1.47mmol/L vs 1.92±0.76mmol/L, P<0.01]. In the condition of serious blood lipid disorders, the ApoE-/- rats had been in a state of inflammation [CRP: 4.66±0.57mg/L vs 0.39±0.21mg/L, P<0.05; white blood cell: (21.79±5.10)×109/L vs (14.82±2.41)×109/L, P<0.01; neutrophil: (9.28±3.35)×109/L vs (2.10±0.96)×109/L, P<0.01]. HE and Masson staining showed that obvious hyperplasia formed and collagen fibers deposited slightly in the two groups. Oil red O staining revealed the obvious hyperplastic plaques in ApoE-/- rats, but a negative result in SD rats. Immunofluorescence staining showed the significant positive CD68 and weak positive α-SMA in the plaque of ApoE-/- rats, while in SD rats the positive α-SMA was pointed out with no CD68 coloration. Conclusion The atherosclerosis model of ApoE-/- rat may be quickly established by short-term high-fat diet combined with balloon injury.

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