核受体视黄醇X受体α对高脂喂养的载脂蛋白E基因敲除小鼠Wnt、β-连环蛋白的调节作用及对动脉粥样硬化的影响  

The effects of retinoid X receptor α agonist bexarotene on Wnt and β-catenin expression and arteriosclerosis in apolipoprotein E knockout mice

作　　者：祝江[1] 张美金 许昌声[1] 林金秀[1] 柴大军[1] ZHU Jiang;ZHANG Mei-jin;XU Chang-sheng;LIN Jin-xiu;CHAI Da-jun(Department of Cardiology,The First Affiliated Hospital of Fujian Medical University,Fujian Hypertension Research Institute,Fuzhou Fujian 350005,China;不详)

机构地区：[1]福建医科大学附属第一医院心血管内科,福建福州350005

出　　处：《中华高血压杂志》2020年第12期1171-1178,共8页Chinese Journal of Hypertension

基　　金：福建医科大学苗圃科研基金资助项目(2015MP023);福建省中青年教师教育科研项目(JAT170236);福建省卫生教育联合攻关计划项目(WKJ2016-2-10)。

摘　　要：目的观察视黄醇X受体(RXR)激动剂蓓萨罗丁(Bex)对高脂喂养的载脂蛋白E(APOE)基因敲除(APOE-/-)小鼠胸主动脉Wnt、β-连环蛋白(β-catenin)的调节作用及对动脉粥样硬化的影响。观察核受体RXRα对血小板源生长因子(PDGF)诱导的平滑肌细胞增殖迁移与Wnt-β-catenin信号转导的关系,探讨核受体RXRα介导Wnt-β-catenin通路对动脉粥样硬化的影响及机制。方法动物实验:8只C57小鼠作为正常对照组,24只APOE-/-小鼠随机分为3组(每组8只):APOE-/-组,APOE-/-+高脂组(高脂喂养APOE-/-小鼠建立动脉硬化模型),APOE-/-+高脂+Bex10组[蓓萨罗丁10 mg/(kg·d)灌胃,干预时间3个月]。检测各组血脂水平,HE染色图像分析测定胸主动脉斑块面积,免疫印迹法检测小鼠胸主动脉Wnt、β-catenin的蛋白表达。细胞实验:培养小鼠胸主动脉平滑肌细胞,(1)分别予PDGF 0、1、3、10、30 ng/mL对平滑肌细胞干预24 h,Western blot方法检测Wnt、β-catenin蛋白表达。(2)PDGF(10 ng/mL)及RXRα激动剂9-顺式维甲酸(9 cis-RA)10-7 mol/L干预24 h,免疫杂交法检测细胞Wnt、β-catenin蛋白水平。(3)PDGF(10 ng/mL)、RXRα激动剂9-cis-RA 10-7 mol/L及β-catenin抑制剂FH535(10-6 mol/L)干预24 h后,5-溴脱氧尿嘧啶核苷(BrdU)渗入法测细胞增殖水平,干预4 h,膜迁移法检测细胞迁移。结果与C57组相比,APOE-/-组血清三酰甘油、总胆固醇、低密度脂蛋白胆固醇(LDL-C)水平升高(均P<0.01),APOE-/-+高脂组三者水平进一步增加(P<0.01);APOE-/-+高脂组与APOE-/-+高脂+Bex10组血清总胆固醇、LDL-C水平差异无统计学意义(P>0.05)。与C57组相比,APOE-/-组胸主动脉Wnt、β-catenin蛋白水平升高,胸主动脉斑块面积显著增加[(64.29±9.87)比0μm2,P<0.05];与APOE-/-+高脂组相比,APOE-/-+高脂+Bex10组Wnt、β-catenin蛋白水平降低,胸主动脉斑块面积降低[(124.09±15.42)比(218.86±15.79)μm2,P<0.05]。与PDGF 0 ng/mL相比,PDGF 10 ng/mL干预后平滑肌细胞Wnt、β-catenin蛋�Objective To explore the effects and the possible mechanisms of retinoid X receptor α(RXRα) agonist bexarotene on Wnt and β-catenin and arteriosclerosis in apolipoprotein E knockout(ApoE-/-) mice. Methods In animal experiments, eight C57 mice were used as normal control group(NCG), and twenty-four ApoE-/- mice were randomly divided into 3 groups: ApoE-/- group, ApoE-/-+high fat chow group(to establish atherosclersis mouse model) group and ApoE-/-+high fat chow+Bex10 gavage group [intervention with bexarotene 10 mg/(kg·d) for 3 months]. Blood lipid levels were detected. Patch area in thoracic aorta was measured by HE staining. Wnt and β-catenin protein levels were detected by immunoblotting. For in vitro experiments, thoracic aorta vascular smooth muscle cells(VSMC) were cultured, and experiments were carried out as following: the Wnt and β-catenin protein level of cells was detected after intervention with platelet derived growth factor(PDGF)(0, 1, 3, 10, 30 ng/mL) for 24 hours, or after intervention with PDGF(10 ng/mL) and RXRα agonist 9-cis-retinoic acid(9 cis-RA) 10-7 mol/L for 24 hours. The proliferation level of VSMC was measured by 5-bromodeoxyuridine(BrdU) infiltration method after intervention with PDGF(10 ng/mL), 9 cis-RA 10-7 mol/L and β-catenin inhibitor FH535(10-6 mol/L) for 24 hours. Cell migration was measured by membrane migration method after intervention with PDGF(10 ng/mL), 9-cis-RA 10-7 mol/L and FH535(10-6 mol/L) for 4 hours. Results Compared with C57 BL/6 mice, triglyceride, total cholesterol, and low-density lipoprotein cholesterol(LDL-C) levels were increased in ApoE-/- mice and increased further in ApoE-/-+ high fat chow mice. Compared with C57 BL/6 group, patch area in thoracic aorta [(64.29±9.87) vs 0 μm2, P<0.05] and Wnt and β-catenin protein levels were increased in ApoE-/-mice. Compared with ApoE-/-+ high fat chow group, patch area in thoracic aorta [(124.09±15.42) vs(218.86±15.79)μm2, P<0.05], Wnt and β-catenin protein levels decreased in ApoE-/-+high fat chow+Bex10 gr

关 键 词：视黄醇X受体 Wnt蛋白 Β-连环蛋白 脂代谢 血管平滑肌细胞 增殖 迁移 动脉粥样硬化 

分 类 号：R543.5[医药卫生—心血管疾病]