硼替佐米对血管紧张素Ⅱ诱导的ApoE^(-/-)小鼠腹主动脉瘤形成的影响  

Effect of Bortezomib on Angiotensin Ⅱ-induced Abdominal Aortic Aneurysm in ApoE^(-/-) Mice

作　　者：李方达[1] 李汇华[2,3] 田翠[4] 聂皓[1] 郑月宏[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院血管外科,北京100730 [2]大连医科大学附属第一医院心血管病研究所,辽宁大连116011 [3]大连医科大学公共卫生学院慢性非传染性疾病预防控制中心,辽宁大连116044 [4]首都医科大学基础医学院生理学与病理生理学系,北京100069

出　　处：《中国医学科学院学报》2017年第1期120-127,共8页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(81270390;81470586);国家重点基础研究发展计划(973计划)(2012CB517802);长江学者计划~~

摘　　要：目的研究蛋白酶体抑制剂硼替佐米(BTZ)对血管紧张素Ⅱ(AngⅡ)诱导的ApoE基因敲除(ApoE^(-/-))小鼠腹主动脉瘤(AAA)形成的炎性机制的影响。方法 8~10周龄雄性ApoE~(^(-/-))小鼠40只,喂养1周适应后随机分为Sham组、BTZ组、AngⅡ组和AngⅡ+BTZ组4组,每组10只。AngⅡ组采用皮下埋植AngⅡ缓释泵[1000 ng/(min·kg)]复制动物AAA模型,BTZ组和AngⅡ+BTZ组同时给予蛋白酶体抑制剂BTZ腹腔注射50μg/kg每周2次。28 d后取材测量统计腹主动脉最大直径及AAA发生率。采用HE染色观察组织中炎症反应情况,免疫组织化学染色及流式细胞分析技术检测组织中T淋巴细胞数目,q PCR检测组织中黏附因子(ICAM-1)的mRNA表达,Western blot检测组织中核转录活化因子信号通路(NF-κB)表达变化。结果 Sham组、BTZ组、AngⅡ组和AngⅡ+BTZ组小鼠的腹主动脉最大直径平均值分别为(1.00±0.01)、(0.99±0.01)、(1.50±0.13)和(1.20±0.04)mm(F=8.959,P=0.000),其中,AngⅡ组明显高于Sham组(P=0.000)和AngⅡ+BTZ组(P=0.015)。AngⅡ组、AngⅡ+BTZ组和Sham组的AAA发生率分别为60%、17%和0。HE染色结果显示,AngⅡ组血管壁厚度及炎症细胞浸润数目较Sham组和AngⅡ+BTZ组明显增加。免疫组织化学染色结果显示,AngⅡ组CD3+T淋巴细胞浸润数为107.9±15.9,明显高于Sham组的0(P=0.000)和AngⅡ+BTZ组的0.8±0.5(P=0.000)。流式细胞检测结果显示,造模1周后,AngⅡ组组织中CD3+T淋巴细胞比例为(13.50±0.69)%,明显高于AngⅡ+BTZ组的(10.40±0.78)%(t=3.009,P=0.040);造模4周后,AngⅡ组的CD3+T淋巴细胞比例为(22.70±0.93)%,明显高于AngⅡ+BTZ组的(15.10±0.97)%(t=5.654,P=0.005)。q PCR检测结果显示,Sham组、BTZ组、AngⅡ组和AngⅡ+BTZ组ICAM-1的mRNA表达水平相比于Sham组升高比例分别为1.00±0.15、0.34±0.03、1.93±0.54和0.83±0.08(F=6.797,P=0.001),其中,AngⅡ组明显高于Sham组(P=0.011)和AngⅡ+BTZ组(P=0.009)。Western blot检测结果显示,AngⅡ组NF-κB核转录抑�Objective To investigate the role of proteasome inhibitor bortezomib（ BTZ） in inflammatory response in abdominal aortic aneurysm（ AAA） formation induced by angiotensin Ⅱ（ Ang Ⅱ）. Methods AngⅡ-induced ApoE^-/-mice AAA models were established. Forty male ApoE^-/-mice（ 8-10-week-old） were randomly and equally divided into four groups： Sham group, BTZ group, Ang Ⅱ group, and Ang Ⅱ ＋ BTZ group. HE staining, immunohistochemical staining, and flow cytometry were used to analyze the inflammatory response.Real-time quantitative polymerase chain reaction（ q PCR） was used to analyze the mRNA expression of intercellular cell adhesion molecule-1（ ICAM-1）. Western blotting was used to analyze the activation of nuclear factor κB signaling（ NF-κB）. Results The mean maximum suprarenal aortic diameter（ Dmax） of Sham group, BTZ group,Ang Ⅱ group, and Ang Ⅱ ＋ BTZ group were（ 1. 00 ± 0. 01）,（ 0. 99 ± 0. 01）,（ 1. 50 ± 0. 13）, and（1. 20 ± 0. 04） mm,respectively（F = 8. 959,P = 0. 000）. The Dmax of Ang Ⅱ group was significantly larger than those of Sham group（ P = 0. 000） and Ang Ⅱ ＋ BTZ group（ P = 0. 015）. The incidence of AAA in Ang Ⅱgroup,Ang Ⅱ ＋ BTZ group,and Sham group were 60%,17%,and 0, respectively. HE staining revealed that the abdominal aortic wall thickening was more severe in Ang Ⅱ group than in Sham group and Ang Ⅱ＋ BTZ group,similar with the infiltration of inflammatory cells. Immunohistochemical staining demonstrated that the CD3＋T lymphocyte count was significantly higher in Ang Ⅱ group than in Sham group（107. 9 ± 15. 9 vs. 0,P = 0. 000） and Ang Ⅱ＋ BTZ group（107. 9 ± 15. 9 vs. 0. 8 ± 0. 5,P = 0. 000）. Flow cytometry also demonstrated that the proportion of the CD3^＋T lymphocytes of the Ang Ⅱ group [（13. 50 ± 0. 69） %] was significantly higher than that in the Ang Ⅱ＋ BTZ group [（10. 40 ± 0. 78） %] at week 1（ t = 3. 009,P = 0. 040）,and the proportion of the CD3^＋T lymphocytes of

关 键 词：腹主动脉瘤 蛋白酶体 硼替佐米 炎症 核转录活化因子信号通路 

分 类 号：R363.21[医药卫生—病理学]