颈总动脉内膜损伤后的自噬现象  

The dynamic changes of autophagy in carotid artery after balloon injury in rats

作　　者：叶兰香[1] 余剑[1,2] 陈歆然[1] 龚琼[3] 丁乔[1] 廖松洁[1,2] 

机构地区：[1]中山大学附属第一医院神经内科,广州510080 [2]广东省重大神经疾病诊治研究重点实验室,卫生部国家临床重点专科,教育部国家重点专科 [3]广东省第二人民医院

出　　处：《中国神经精神疾病杂志》2014年第7期409-413,共5页Chinese Journal of Nervous and Mental Diseases

基　　金：国家自然科学基金青年科学基金项目(编号:81100871);国家自然科学基金面上项目(编号:81371276)510080;中大青年教师培育基金项目(编号:13ykpy16)

摘　　要：目的观察颈总动脉内膜损伤后自噬的动态变化。方法 40只雄性SD大鼠利用简单随机抽样法分为假手术组(n=20)和模型组(n=20)。假手术组依次分离颈部皮肤、皮下组织,暴露左侧颈总动脉后直接缝合皮肤,模型组行左侧颈总动脉内膜剥脱术。假手术组及模型组在3 d、7 d分别用western blot、免疫荧光检测颈总动脉中Beclin-1、微管相关蛋白1轻链3(microtubule associated protein 1 light chain 3,LC3)的表达;并在相同的时间点用western blot检测颈总动脉中P62的表达。结果 western blot结果显示,与假手术组相比,模型组Be-clin-1及LC3的表达于3 d、7 d增高,其中第7 d增高明显,自噬底物P62的表达于3 d、7 d下降,其中7 d下降显著。免疫荧光结果显示,与假手术组相比,模型组Beclin-1在颈总动脉中膜及内膜总的表达增高(第3 d 18.60±1.34;6.40±0.55,P=0.000,n=5;7 d 27.6±2.19;6.40±0.55,P=0.000,n=5);模型组LC3在颈总动脉中膜及内膜总的表达增高(第3d 10.60±1.52;3.00±0.71,P=0.000,n=5;第7d 21.20±2.49;3.00±0.71,P=0.000,n=5)。结论模型组中调控自噬的Beclin-1蛋白以及自噬体形成所必需的LC3的表达增高,均表明自噬被激活,并于不同时间点呈动态变化,推测其可能参与新生内膜形成。Objective To investigate the dynamic levels of autophagy after intimal injury of carotid artery. Meth-ods In this study ,40 male SD rats were randomly assigned to operated（n=20）and control groups（n=20）. Balloon inju-ry was induced in the left carotid artery in operated groups .Rats in control groups just received carotid artery exposure without injury. Western blot was used to detect the levels of Beclin-1, LC3 and p62 at the third and seventh days. Immu-nofluorescence was used to examine the expression of Beclin-1 and LC3 at the third and seventh days. Results The ex-pression levels of Beclin-1 and LC3 were increased while the levels of P62 were decreased at the third and seventh days after carotid balloon injury. Beclin-1 and LC3 were present in neointima and medintima. The numbers of both Beclin-1 positive cells and LC3 positive cells were increased at the third and seventh days after carotid injury. The numbers of Be-clin-1 positive cells were 18.60 ± 1.34 in neointima and 6.40 ± 0.55 in medintima at third day, （27.6 ± 2.19 in neointima and 6.40±0.55 in medinitima at the seventh day,（all P=0.000,n=5）. The numbers of LC3 positive cells were 10.60±1.52 in neointima and 3.00 ± 0.71 in medintima at third day, （P=0.000,n=5;at the seventh day 21.20 ± 2.49;3.00 ± 0.71,P=0.000,n=5）. Conclusions This study domenstrates that autophagy was activated after carotid injury and the chang is dy-namic, which may contribute to neointima formation.

关 键 词：颈动脉新生内膜 自噬 LC3 BECLIN-1 

分 类 号：R743.3[医药卫生—神经病学与精神病学]