β3肾上腺素能受体活化抑制慢性间歇低氧诱导的动脉粥样硬化进展的机制  

Impact and Related Mechanisms of Activation ofβ3 Adrenergic Receptor on Atherosclerosis Induced by Chronic Intermittent Hypoxia in Apo E Knockout Mice

作　　者：索旻[1] 王越 刘梦如 刘倍倍 陈蕾蕾 王悦[2] 王志强 张慧娜[3,4] 吴小凡 SUO Min;WANG Yue;LIU Mengru;LIU Beibei;CHEN Leilei;WANG Yue;WANG Zhiqiang;ZHANG Huina;WU Xiaofan(Coronary Heart Disease Center,Beijing Anzhen Hospital,Capital Medical University,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院冠心病中心,北京市100029 [2]首都医科大学附属北京安贞医院心内科,北京市100029 [3]北京市心肺血管疾病研究所 [4]上气道功能障碍相关心血管疾病研究室

出　　处：《中国循环杂志》2021年第12期1237-1244,共8页Chinese Circulation Journal

基　　金：北京市百千万人才工程(2020A39);北京市教育委员会科技计划重点项目(KZ202010025045);国家自然科学基金(82071573、81670317、81470492)。

摘　　要：目的:探讨在慢性间歇低氧(CIH)促进动脉粥样硬化进展的过程中,β3肾上腺素能受体(β3 AR)活化对动脉粥样硬化进展的影响及机制。方法:将15只6~7周龄雄性载脂蛋白E基因敲除(ApoE-/-)小鼠按照随机数表法等分为3组:常氧组、低氧组与低氧+米拉贝隆组。对照组给予高脂饲料喂养12周;低氧组和低氧+米拉贝隆组于高脂饲料喂养4周后每日进入低氧箱8 h,持续8周(箱内氧浓度在5%~21%之间循环,每个循环180 s),低氧+米拉贝隆组小鼠进入低氧箱的同时还每日给予10 mg/kg的米拉贝隆灌胃。12周时收取小鼠主动脉组织,苏木素-伊红染色检测主动脉根部粥样硬化斑块面积;CD68免疫组织化学染色检测斑块内巨噬细胞;用ABTS法检测血清总抗氧化能力;免疫组织化学染色检测斑块内p22phox表达;超氧化物阴离子荧光探针检测活性氧;转录组测序方法筛选各组小鼠主动脉组织氧化应激相关的差异表达基因。培养RAW264.7细胞,通过细胞低氧培养箱(氧浓度在5%~21%之间循环,每个循环1 h)建立CIH模型,分为低氧组和低氧+米拉贝隆组(米拉贝隆0.5μmol/L),48 h后用ABTS法检测细胞总抗氧化能力;用免疫印迹法检测巨噬细胞β3 AR、p22phox表达水平。结果(:1)低氧组较常氧组和低氧+米拉贝隆组小鼠的主动脉根部斑块面积占动脉壁表面积的百分比更高[(27.88±5.83)%vs.(19.84±5.13)%(;27.88±5.83)%vs.(16.33±3.60)%,P均<0.05],斑块内CD68阳性面积百分比更高[(10.63±3.18)%vs.(5.31±2.77)%(;10.63±3.18)%vs.(5.20±1.40)%,P均<0.05],斑块内β3 AR阳性面积百分比更高[(5.06±0.44)%vs.(2.99±1.56)%(;5.06±0.44)%vs.(2.61±0.95)%,P均<0.05],p22phox阳性面积百分比更高[(4.82±0.85)%vs.(1.29±0.27)%(;4.82±0.85)%vs.(0.94±0.63)%,P均<0.05];各组小鼠斑块内p22phox阳性面积与β3 AR阳性面积呈正相关(r=0.90,P<0.05)。(2)低氧组较常氧组和低氧+米拉贝隆组小鼠颈动脉组织活性氧阳性面积百分比更高[(29.Objectives:To investigate the impacts and related mechanisms of activatedβ3 adrenergic receptor(β3 AR)on atherosclerosis induced by chronic intermittent hypoxia(CIH).Methods:Male ApoE knockout mice(6-7 weeks)were randomly divided into three groups(n=5 each):normoxia group,hypoxia group and hypoxia+mirabegron group.The normoxia group was fed with high-fat diet for 12 weeks,the hypoxia group and the hypoxia+mirabegron group were fed with high-fat diet for 4 weeks and then entered the hypoxic chamber(the oxygen concentration in the chamber was between 5%and 21%,each cycle was 180 seconds)for 8 hours per day for 8 weeks.Mice in the hypoxia+mirabegron group received 10 mg/kg mirabegron by gavage daily for 8 weeks.Aortic tissue was collected at 12 weeks,and the area of atherosclerotic plaque in aortic root was detected by HE staining.CD68 immunohistochemical staining was performed to detect macrophages in the plaque.Immunohistochemical staining was performed to detect the expression of p22 phox in plaques.The total antioxidant capacity of serum was determined by ABTS.In vitro,RAW264.7 cells were cultured and CIH model was established by cell hypoxia incubator(oxygen concentration was between 5%and 21%,each cycle was 1 hour).The cells were divided into hypoxia group and hypoxia+mirabegron group(mirabegron 0.5μmol/L).After 48 hours,the expression levels ofβ3 AR and p22 phox in macrophages were detected by Western blot,and the total antioxidant capacity of cells was detected by ABTS method.Results:(1)The aortic root atherosclerosis plaque area of mice([27.88±5.83]%vs.[19.84±5.13]%,[27.88±5.83]%vs.[16.33±3.60]%,all P<0.05),CD68 positive area of mice([10.63±3.18]%vs.[5.31±2.77]%,[10.63±3.18]%vs.[5.20±1.40]%,all P<0.05),β3 AR positive area of mice([5.06±0.44]%vs.[2.99±1.56]%,[5.06±0.44]%vs.[2.61±0.95]%,all P<0.05),and p22 phox positive area of mice([4.82±0.85]%vs.[1.29±0.27]%,[4.82±0.85]%vs.[0.94±0.63]%,all P<0.05)were significantly larger in the hypoxia group than in the normoxia group and hypoxia+mirab

关 键 词：慢性间歇低氧 动脉粥样硬化 Β3肾上腺素能受体 氧化应激 

分 类 号：R541.4[医药卫生—心血管疾病]