摘要
背景:易损斑块是致急性心、脑血管缺血事件的病理基础,建立合适的动物易损斑块模型以供研究对心脑血管疾病的防治工作具有重要的指导意义,但目前尚无公认的构建易损斑块模型的方法。目的:探讨免疫刺激能否协同液氮冷冻损伤血管构建大鼠动脉粥样硬化易损斑块模型,以期能够复制出能广泛应用的动脉粥样硬化模型。方法:雄性SD大鼠40只,经适应性喂养1周后将其随机分为4组,即对照组、高脂组、液氮组、免疫刺激+液氮组。对照组予以常规饲料,其他3组予以高脂饲料喂养,2周后各组大鼠腹腔一次性注射维生素D3注射液;对照组及高脂组大鼠仅做假手术处理,不进行液氮及免疫刺激,术后分别继续予以常规饲料及高脂饲料喂养10周;液氮组及液氮+免疫刺激组施以液氮冷冻损伤术,术后继续予以高脂饲料饲养10周,期间液氮+免疫刺激组大鼠皮下注射牛血清白蛋白(3次/周,共计3周)及腹腔注射卵清白蛋白(间隔3 d注射,共计5次)。造模10周后,取血检测大鼠血脂指标、炎症及氧化应激损伤指标、血红素氧合酶1蛋白表达量;取靶血管段观察粥样斑块的病理变化。实验方案经西南医科大学实验动物伦理委员会批准(批准号为IACUC:20170917006)。结果与结论:①与对照组大鼠相比较,高脂组大鼠胆固醇水平、炎症及氧化应激指标、血红素氧合酶1蛋白表达量以及镜下组织病理学检查均无明显变化(P>0.05);与对照组及高脂组相比较,液氮组、液氮+免疫刺激组大鼠胆固醇水平、炎症及氧化应激指标、血红素氧合酶1蛋白表达量均明显升高(P<0.05),镜下组织病理学检查也提示有明显斑块,且液氮+免疫刺激组升高趋势更为显著(P<0.05);与液氮组相比较,液氮+免疫刺激组大鼠血脂、炎症及氧化应激指标、血红素氧合酶1蛋白表达量均明显升高(P<0.05),镜下组织病理学检查提示粥样斑块较液氮组更为明显;②结果说明,免疫刺激能协同液氮冷冻损伤血管加速大鼠颈总动脉粥样硬化易损斑块模型形成,为动脉粥样硬化性疾病的干预实验奠定基础。
BACKGROUND:Vulnerable plaque is the pathological basis of acute cardiac and cerebrovascular ischemic events.Establishing a suitable animal vulnerable plaque model is of great significance for guiding the prevention and treatment of cardiovascular and cerebrovascular diseases.However,there is no well-established method for constructing the vulnerable plaque model.OBJECTIVE:To investigate whether immunostimulation can cooperate with liquid nitrogen freezing to establish an atherosclerotic vulnerable plaque model in rats,aiming to replicate the widely applicable atherosclerotic model.METHODS:Forty male Sprague-Dawley rats were randomly divided into four groups after 1 week of adaptive feeding,namely,control group,high-fat group,liquid nitrogen group,immunostimulation+liquid nitrogen group.After 2 weeks of conventional diet and special high-fat diet,respectively,the rats in the control group and the other groups were given a single dose of vitamin D3 injection through the abdominal cavity.Rats in the control and high-fat diet groups were subjected to sham operation with no immunostimulation and liquid nitrogen freezing,and given routine diet and special high-fat diet for 10 weeks respectively.Rats in the liquid nitrogen group and immunostimulation+liquid nitrogen group were given liquid nitrogen freezing,followed by 10 weeks of high-fat diet.During this period,the rats in the immunostimulation+liquid nitrogen group were given subcutaneous injection of bovine serum albumin(3 times per week,for 3 weeks)and intraperitoneal injection of ovalbumin(3 days interval,5 times in total).At 10 weeks after modeling,blood samples were taken to detect blood lipid index,inflammation and oxidative stress injury index,and heme oxygenase-1(HO-1)protein expression in rats.The target blood vessel segments of each group were executed for hematoxylin-eosin staining,and the pathological changes of atherosclerotic plaques were observed under light microscopy.The study protocol was approved by the Animal Ethics Committee of Southwest Medical University,with an approval No.IACUC:20170917006.RESULTS AND CONCLUSION:Compared with the control group,there were no significant changes in cholesterol level,inflammatory and oxidative stress index,HO-1 protein expression and histopathological examination in the high-fat group(P>0.05).Compared with the control and high-fat groups,the levels of serum lipids,inflammatory and oxidative stress indicators and HO-1 protein expression in the liquid nitrogen and immunostimulation+liquid nitrogen groups were significantly increased(P<0.05).Histopathological examination also indicated that there were obvious plaques,and the increasing trend in the immunostimulation+liquid nitrogen group was more significant(P<0.05).Compared with the liquid nitrogen group,the levels of serum lipids,inflammatory and oxidative stress indexes and HO-1 protein expression in the immunostimulation+liquid nitrogen group were significantly increased(P<0.05),and histopathological examination also showed more obvious atherosclerotic plaques.Therefore,immunostimulation could accelerate the formation of rat common carotid atherosclerotic vulnerable plaque model in cooperation with liquid nitrogen freezing injury,which lays a foundation for the intervention experiment of atherosclerotic disease.
作者
魏刚
高尚远
张英
黄维义
Wei Gang;Gao Shangyuan;Zhang Ying;Huang Weiyi(Department of Cardiology,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Department of Cardiology,Dujiangyan People’s Hospital,Dujiangyan 611800,Sichuan Province,China;Functional Laboratory of Southwest Medical University,Luzhou 646000,Sichuan Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2020年第35期5656-5661,共6页
Chinese Journal of Tissue Engineering Research
基金
四川省科技厅-泸州市科技局-泸州医学院联合基金项目(14JC0051),项目负责人:张英
泸州市科技局基金项目(2013LZLY-J15),项目负责人:黄维义。
关键词
实验
模型
鼠
免疫
冷冻
动脉粥样硬化
血管构建
颈总动脉
experiment
model
rat
immune
freezing
atherosclerosis
vascular construction
common carotid artery
