化合物48/80对载脂蛋白E基因敲除小鼠主动脉斑块的影响  

Effect of Compound 48/80 on Thoracic Aortic Plaque in Apolipoprotein E-knock out Mice

作　　者：王双 [1] 杨永宗 [2] 唐雅玲 [2] 唐朝克 [2] 易光辉 [2] 孙玉慧 [2] 叶旭 [2] 许增祥 [2] 彭旷 [2] 彭倩 [2] 万载阳 [1] 

机构地区：[1]中南大学湘雅医学院病理生理系,湖南省,长沙市,410078,南华大学心血管病研究所,湖南省,衡阳市,421001 [2]南华大学心血管病研究所,湖南省,衡阳市,421001

出　　处：《中国动脉硬化杂志》2005年第3期292-296,共5页Chinese Journal of Arteriosclerosis

基　　金：湖南省重点科技项目(01SSY1003)资助

摘　　要：目的目前,肥大细胞与动脉粥样硬化的关系研究主要是病理学观察和细胞学实验。基于此种现状,本课题拟用化合物48/80探讨肥大细胞脱颗粒与载脂蛋白E基因敲除小鼠主动脉斑块的关系。方法40只10周龄载脂蛋白E基因敲除小鼠给予高脂高胆固醇饲养至16周龄,随机分为2组(n=20),实验组给予腹腔注射化合物48/80(0.5mg/kg),隔日一次,共4次;对照组腹腔注射Dhank’s。第4次注射后半小时,安乐死处死动物,摘眼球取血,分离血清,采用酶法测定血清脂质含量,采用比色法测定血清类胰蛋白酶活性浓度。原位灌流固定,取主动脉置10%中性缓冲福尔马林中固定,石蜡包埋,连续切片,HE染色、甲苯胺蓝染色以及α平滑肌肌动蛋白、Mac3、VEcadherin免疫组织化学染色。HMIAs2000高清晰度彩色医学图文分析系统进行图像分析。结果实验组血清类胰蛋白酶活性浓度明显高于对照组(0.57±0.13u/L比0.36±0.10u/L),但两组血清脂质含量无明显差异。实验组与对照组相比,主动脉外膜脱颗粒肥大细胞百分率升高(80.6%±17.8%比13.5%±4.1%,P<0.05),主动脉斑块最大横截面积增大[(1.25±0.36)×104μm2比(0.79±0.24)×104μm2,P<0.05],斑块内α平滑肌肌动蛋白阳性细胞百分率下降(36.2%±14.9%比69.7%±31.3%,P<0.05)斑块内Mac3阳性细胞百分率升高(58.6%±17.3%比28.5%±16.4%,P<0.05),而斑块内膜VEcadherin平均光密度减小(48±8比65±10,P<0.05)。结论化合物48/80促使肥大细胞脱颗粒,并使载脂蛋白E基因敲除小鼠主动脉斑块最大横截面积增大、斑块内Mac3阳性细胞百分率升高、α平滑肌肌动蛋白阳性细胞百分率下降、斑块内膜VEcadherin平均光密度减小。Aim So far, research on the relationship of mast cells, atheroselerosis and angiogencsis is mostly pathological observation and cytology experiments. Based on the status, compound 48/80 and apolipoprotein E-knockout mice were used to investigate the effect of mast cell degranulation on plaque. Methods 40 apoE-/- mice were fed a Western-type diet from 10wecks old. At 16wecksold, mice were divided into 2 groups and treated for 8 days as follows ： group I, mice were intraperitoncal injected with compound 48/80, 0.5 mg/kg, every other day; and group 2, control mice were intraperitoncal injected with buffer. After half an hour of the fouth injection, euthanasia, blood was collected from the orbit for measurement of plasma lipids and tryptase, mice were in situ peffusion fixed with 10% neutral buffered formalin and aortas were removed. Then aortas were fixed in neutral buffered formalin, and embedded in paraffin. Sections were routinely stained with hematoxylin and eusin. Cotresponding sections on separate slides were stained immunohistochemically with antibodies against a macrophage-specific antigen （Mac3）, α-smooth muscle actin, VE-cadherin. The images were analyzed with a Olympus BX51 microscope and HMIAS2000 software to distinguish the maximum cross section area of aorta plaque and expression of Mac3, α-smooth muscle actin, VE-cadherin between the two groups. Results Tryptase levels were 0.54±0.29 versus 0.36±0.13 mmol/L in the Compound 48/ 80 and control groups, respectively （P〈0.05）. There was no difference on serum lipids content. Compound48/80 group were compared with control group, the maximum cross section area of aorta plaque was significantly increased [（1. 25±0.36）×10^4μm^2 VS（0.79±0.24）×10^4μm^2 in control group, P〈0.05],the percent of Mac3 ＋ cell was increased （58.6%±17.3% vs 28.5%±16.4%, P〈0.05）, the percent of α- muscle actin cell was reduced （ 36.2% ±14.9% vs 69.7 %±31.3 %, P〈0.05 ） and the MOD of VE-eadherin of aorta endothelium was reduced （48±8

关 键 词：病理学与病理生理学 肥大细胞 主动脉斑块 载脂蛋白E基因敲除小鼠 化合物48/80 

分 类 号：R363[医药卫生—病理学]