重组腺病毒介导脂联素基因对ApoE^-/-小鼠动脉粥样硬化的作用机制  

Mechanism of the effect of r Ad-APN gene on atherosclerosis in ApoE^(-/-) mice

作　　者：王雪梅[1] 魏琴[1] 张春[1] 姜涛[1] 段明军[1] 杨毅宁[1] 

机构地区：[1]新疆医学动物模型研究重点实验室,新疆医科大学第一附属医院临床医学研究院,乌鲁木齐830011

出　　处：《中国实验动物学报》2016年第2期175-182,共8页Acta Laboratorium Animalis Scientia Sinica

基　　金：新疆维吾尔自治区自然科学基金青年基金项目(2015211C095);新疆重大疾病医学重点实验室-省部共建国家重点实验室培育基地(2010DS890294)开放课题(SKLIB-XJMDR-2014-16)

摘　　要：目的研究腺病毒介导的脂联素(APN)过表达对Apo E-/-小鼠动脉粥样硬化(AS)的抑制作用和对NF-κB信号通路的影响。方法将12周龄雄性Apo E-/-小鼠120只分为空载腺病毒对照组和脂联素干预组,每组60只。在3个不同时间点(0天、4周、8周)处死小鼠收集组织。全自动生化仪检测血脂指标;ELISA法测定血清APN浓度;油红O染色法检测小鼠主动脉血管组织的病理学变化;Masson染色法检测进展性斑块区的胶原含量和纤维帽厚度变化;免疫荧光法测定小鼠主动脉血管APN和NF-κB p65蛋白的表达;免疫印迹法检测主动脉血管APN、NF-κB p65核蛋白和炎症因子的表达。结果 APN过表达抑制了Apo E-/-小鼠动脉粥样硬化斑块形成。与对照组相比,脂联素干预组的动脉粥样硬化病理损伤面积减少(P<0.01),动脉粥样硬化损伤程度降低(P<0.001),总体油红O染色测定血管表面损伤百分比,4周时为(27.78±8.64)vs(33.02±5.18)%;8周时为(31.58±5.87)vs(52.16±5.79)%。脂联素减缓了高脂饮食导致的小鼠血清中TC(P<0.001)、TG(P<0.001)、LDL-C(P<0.001)的浓度增加,使血脂水平趋向正常化。随着血清脂联素浓度增加,可以阻遏NF-κB通路的激活,抑制NF-κB p65核蛋白和炎症因子的表达。结论脂联素通过抑制NF-κB通路激活来减轻AS的炎症反应。Objective Atherosclerosis（ AS） is a common pathological basis of cardiovascular diseases. Adiponectin（ APN） has been shown to have an anti-AS effect,and the underlying mechanisms,however,are largely unknown. Nuclear transcription factor κB（ NF-κB） has also been regarded as a proatherogenic factor,mainly because of its regulation of a variety of the proinflammatory genes linked to AS. It is hypothesized that the inhibitory effects of APN on AS is through the inhibition of NF-κB signaling pathway. The aim of this study was to test the hypothesis via investigation and validation of the inhibitory effect of APN on AS in Apo E^- /-mice,and to delineate the roles of NF-κB signaling pathway in modulating the APN effect on AS in vivo. Methods APN overexpression in Apo E^- /-mice were mediated by transfecting adenovirus bearing a vector encoding for APN and enhanced green fluorescent protein（ Ad-APN-e GFP）. The AS in Apo E^- /-mice was induced by feeding a high-fat diet. To validate the inhibitory effect of the adenovirus mediated APN overexpression on AS inthe ApoE^- /-mice. 120 male ApoE^- /-mice aged 12 weeks were randomly and evenly assigned into two groups（ 60 mice per group）,and were fed with a high-fat diet to induce AS. At 0 day,2,4,and 6 weeks of high-fat diet feeding. The 2groups of mice were injected intravenously in the tail with either 100 μL（ 3. 0 × 10^8 p. f. u） of Ad-eG FP virus（ control group） or the same amount of Ad-APN-eG FP virus（ APN group）. Blood samples and aortic tissues were taken at 0 day,4,and 8 weeks of high-fat diet feeding. For the blood samples,FABA was used to analyze the concentrations of blood lipids and ELIZA was used to test the concentrations of serum APN. For the aortic tissues,oil red O staining was used to detect the surface lesion percentage. Masson staining was used to evaluate the collagen content and fibrous cap thickness of the plaque area. Immunofluorescence method was used to detect APN and NF-κB p65 expression. Western blot was used to

关 键 词：脂联素 动脉粥样硬化 APO E^-/-小鼠 炎症 NF-ΚB信号通路 

分 类 号：Q95-33[生物学—动物学]