CAJ | 학술논문

目的探讨主动脉夹层小鼠血管壁中微管相关蛋白1轻链3(Microtubule-associated protein 1 light chain 3,LC3)、Beclin1及骨桥蛋白(Osteopontin,OPN)的表达及意义.方法 (1)急性主动脉夹层模型的建立:3周龄FVB小鼠随机分为β-氨基丙腈(β-aminopropionitrilemonofumarate,BAPN)联合人血管紧张素-Ⅱ(Angiotensin-Ⅱ,Ang-Ⅱ)组,单BAPN干预组和对照组,分别予以BAPN口服4周后联合Ang-Ⅱ皮下微泵48 h,单用BAPN 4周口服和正常喂养进行干预.(2) LC3、Beclin1及OPN的检测:采用RT-PCR、免疫组织化学染色法对小鼠主动脉血管壁中自噬相关蛋白LC3、Beclin1及合成型血管平滑肌细胞(vascular smooth muscle cell,VSMC)标志蛋白OPN等进行定量和半定量测定.结果 (1)BAPN联合Ang-Ⅱ干预组小鼠夹层发生比例为86.7％,单BAPN组夹层发生比例低于联合组,对照组夹层发生比例为0;(2)急性主动脉夹层小鼠主动脉壁中自噬相关蛋白LC3B、Beclin1及合成型VSMC标志蛋白OPN基因和蛋白含量明显高于正常对照组(P＜0.05),且LC3B、Beclin1及OPN蛋白表达主要定位于血管中层VSMC.结论 BAPN联合Ang-Ⅱ能够成功诱导FVB小鼠发生急性主动脉夹层,且该模型中发生了VSMC表型转化和自噬现象,两者可能具有相关性.
목적 탐토주동맥협층소서혈관벽중미관상관단백1경련3(Microtubule-associated protein 1 light chain 3,LC3)、Beclin1급골교단백(Osteopontin,OPN)적표체급의의.방법 (1)급성주동맥협층모형적건립:3주령FVB소서수궤분위β-안기병정(β-aminopropionitrilemonofumarate,BAPN)연합인혈관긴장소-Ⅱ(Angiotensin-Ⅱ,Ang-Ⅱ)조,단BAPN간예조화대조조,분별여이BAPN구복4주후연합Ang-Ⅱ피하미빙48 h,단용BAPN 4주구복화정상위양진행간예.(2) LC3、Beclin1급OPN적검측:채용RT-PCR、면역조직화학염색법대소서주동맥혈관벽중자서상관단백LC3、Beclin1급합성형혈관평활기세포(vascular smooth muscle cell,VSMC)표지단백OPN등진행정량화반정량측정.결과 (1)BAPN연합Ang-Ⅱ간예조소서협층발생비례위86.7％,단BAPN조협층발생비례저우연합조,대조조협층발생비례위0;(2)급성주동맥협층소서주동맥벽중자서상관단백LC3B、Beclin1급합성형VSMC표지단백OPN기인화단백함량명현고우정상대조조(P＜0.05),차LC3B、Beclin1급OPN단백표체주요정위우혈관중층VSMC.결론 BAPN연합Ang-Ⅱ능구성공유도FVB소서발생급성주동맥협층,차해모형중발생료VSMC표형전화화자서현상,량자가능구유상관성.
Objective To explore the expression of microtubule-associated protein 1 light chain 3 、Beclin1 and osteopontinin aortic tissue of aortic dissection mice and to discuss the relevance.Methods (1) The establishment of aortic dissection model:3 weeks old FVB mice were randomly divided intoβ-aminopropionitrilemonofumarate (BAPN) with Angiotensin-Ⅱ (Ang-Ⅱ) group,BAPN only group and control group.The BAPN-Ang-Ⅱ group was intervened by BAPN orally at first 4 weeks and infused with Ang Ⅱ subcutaneously using a micro-osmotic pump for 48 h.The BAPN only group was intervened by BAPN orallyfor 4 weeks,and the control was given a placebo respectively.(2) Detecting the expression of microtubule-associated protein 1 light chain 3 、Beclin1 and osteopontin:RT-PCR and Immunohistochemistry staining was performed to detect the expression of LC3 B,Beclin1 and OPN in the aortic tissue of aortic dissectionmice.Results (1) the incidence of aortic dissection in the BAPN-Ang-Ⅱ is 86.7％ and the BAPN only group is lower than the collaborategroup.No aortic dissection was found in the control group.(2)It was identified that the autophagy related gene was much highly expressed in the AD mice's aortic tissue verse the normal one.According to immunohistochemistry staining,the autophagy related protein is mainly located in the medial vascular wall.Conclusion These data for the first time demonstrates that autophagy related gene and protein highly expressed in the AAD mouse's aortic tissue and it will provide us an available method and fundamental for the further studying in the influences of autophagy to AAD.