Rev-erbα在血管内皮损伤后内膜增生中的作用  

Role of Rev-erbα in neointima hyperplasia after vascular endothelial injury

作　　者：甘雪晴 侯娟妮 李秀川 孙雄山 杨大春 Gan Xueqing;Hou Juanni;Li Xiuchuan;Sun Xiongshan;Yang Dachun(School of Clinical Medicine,Southwestom Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学临床医学院,泸州646000 [2]西部战区总医院心内科

出　　处：《中华老年心脑血管病杂志》2020年第6期629-634,共6页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(81770299)。

摘　　要：目的探讨Rev-erbα在血管内皮损伤后内膜增生中的作用。方法将雄性野生型C57BL/6J小鼠80只随机分为假手术1组和2组、激动剂1组、抑制剂1组、损伤1组和2组、激动剂2组和抑制剂2组,每组10只,前4组仅行假手术,后4组用机械损伤颈动脉内皮法构建模型,造模后,按分组分别腹腔注射激动剂SR9011或抑制剂SR8278各100 mg/kg,1次/d,4周。用100 nmol/L血管紧张素Ⅱ(AngⅡ)干预小鼠主动脉平滑肌细胞系(MOVAS)24 h诱导体外增殖,特异性小分子干扰(siRNA)转染技术敲低细胞中核苷酸结合寡聚化域样受体蛋白3(Nlrp3)水平。将细胞分为对照1组、2组和3组,AngⅡ1组、2组和3组,SR9011组,SR8278组,AngⅡ+SR8278组,siRNA对照组,si-Nlrp3组,SR8278+si-Nlrp3组,后9组用AngⅡ干预24 h,SR9011组和SR8278组先用5μmol/L的SR9011或SR8278干预12 h;后3组用siRNA敲低Nlrp3水平。分别用qRT-PCR和Western blot测Rev-erbα、Nlrp3 mRNA和蛋白表达,苏木精-伊红染色观察颈动脉内膜增生,分别用CCK-8法和Ki-67免疫荧光术测细胞增殖率。结果与假手术1组比较,内皮损伤1组Rev-erbαmRNA及蛋白表达下降(P<0.01)。与假手术2组比较,内皮损伤2组内膜增生更明显(P<0.01)。与内皮损伤2组比较,激动剂2组内膜/中膜面积比值下降(P<0.01);抑制剂2组内膜/中膜面积比值升高(P<0.01)。与对照1组比较,AngⅡ1组Rev-erbαmRNA和蛋白表达下降(P<0.01)。与AngⅡ2组比较,SR9011组吸光度和阳性细胞率减少,SR8278组吸光度和阳性细胞率明显增加(P<0.05)。与对照2组比较,AngⅡ2组Nlrp3 mRNA和蛋白表达增加(P<0.01);与AngⅡ2组比较,SR9011组Nlrp3 mRNA和蛋白表达降低,SR8278组Nlrp3 mRNA和蛋白表达增加(P<0.05)。si-Nlrp3组较siRNA对照组阳性细胞减少[(0.133±0.012)%vs(0.337±0.023)%,P<0.05];SR8278+si-Nlrp3组较AngⅡ+SR8278组阳性细胞减少[(0.138±0.011)%vs(0.498±0.023)%,P<0.05]。结论Rev-erbα可能通过抑制Nlrp3表达,抑制细胞增殖,抑制血管内皮损伤Objective To study the role of Rev-erbαin neointima hyperplasia after vascular endothelial injury.Methods Eighty male wild C57 BL/6 J mice were randomly divided into sham operation groups 1 and 2,agonist group 1,antagonist group 1,injury groups 1 and 2,agonist group 2 and antagonist group 2(10 in each group).The first 4 groups only underwent sham operation.A model of vascular endothelial injury was established for the latter 4 groups by mechanically injuring the carotid artery.The animals in agonist groups 1 and 2,antagonist groups 1 and 2 were injected with SR9011 or SR8278 respectively(100 mg/kg,once a day)for 4 weeks after the model was established.The MOVAS were treated with 100 nmol/L angiotensinⅡ(AngⅡ)for 24 hours to induce proliferation in vitro.Specific siRNA transfection technique was used to knock down the level of Nlrp3.The MOVAS were divided into control groups 1,2 and 3,AngⅡgroups 1,2 and 3,SR9011 group,SR8278 group,AngⅡ+SR8278 group,siRNA control group,si-Nlrp3 group and SR8278+si-Nlrp3 group.The MOVAS in the latter 9 groups were treated with AngⅡfor 24 hours.The MOVAS in SR9011 group and SR8278 group were treated with SR9011 or SR8278 of 5μmol/L for 12 hours.The Nlrp3 level in the latter 3 groups were knocked down using siRNA.The expressions of Rev-erbαand Nlrp3 were detected by qRT-PCR and Western blot respectively.The carotid artery neointima hyperplasia was detected with HE staining.The proliferation rate of cell was assayed by CCK-8 and Ki-67 immunofluorescence staining.Results The expression levels of Rev-erbαmRNA and protein were significantly lower in endothelial injury group 1 than in sham operation group 1(P<0.01).The neointimal hyperplasia was severer in endothelial injury group 2 than in sham operation group 2(P<0.01).The intima area/media area ratio was lower in agonist group 2 than in endothelial injury group 2(P<0.01)while the intima area/media area ratio was higher in antagonist group 2 than in endothelial injury group 2(P<0.01).The expression levels of Rev-erbαmRNA an

关 键 词：内皮 血管 血管紧张素Ⅱ 肌细胞 平滑肌 核受体亚家族1 D族 成受1 

分 类 号：R363[医药卫生—病理学]