载脂蛋白E基因敲除小鼠动脉粥样硬化早期血管外膜成纤维细胞增殖活性增强  

Increase in proliferation of adventitial fibroblasts at early stage of atherosclerosis in apoE(-/-) mice

作　　者：徐芳[1] 刘颖[2] 石磊[1] 蔡虹静[1] 王蔚琛[1] 胡维诚[1] 

机构地区：[1]滨州医学院病理生理学教研室,山东滨州256603 [2]滨州医学院附属医院,山东滨州256603

出　　处：《中国病理生理杂志》2010年第8期1503-1508,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30470700)

摘　　要：目的:探讨动脉外膜成纤维细胞增殖与早期动脉粥样硬化病灶形成的关系。方法:选择6周龄载脂蛋白E基因敲除[apoE(-/-)]小鼠和野生型C57BL/6小鼠,高脂喂养2、4和10周后,在各个时点处死动物前24 h经腹腔注射5-溴-2-脱氧尿嘧啶(BrdU),后选取升主动脉制备连续切片,通过HE染色观察组织形态学的变化,用免疫组化方法观察不同时点血管外膜及内膜BrdU的表达变化。体外培养高脂喂养2周的apoE(-/-)小鼠和C57BL/6小鼠动脉外膜成纤维细胞,通过BrdU掺入法测定细胞增殖活性,流式细胞术测定细胞周期。结果:体内实验发现apoE(-/-)小鼠高脂喂养2周后,在无可见内膜病灶形成之前,首先在主动脉外膜发现BrdU标记的阳性细胞,之后才在损伤内膜观察到BrdU标记细胞。而C57BL/6小鼠在任何时点都未检测到BrdU标记的细胞。体外实验观察到apoE(-/-)小鼠血管外膜成纤维细胞BrdU标记的细胞数显著多于C57BL/6小鼠(P<0.01),apoE(-/-)小鼠血管外膜成纤维细胞S期及G2/M期所占百分比明显高于对照组(P<0.05)。结论:血管外膜成纤维细胞增殖可能参与早期动脉粥样硬化病灶形成。AIM： To study the relationship between the proliferation of adventitial fibroblasts and the early formation of atherosclerotic lesion.METHODS： ApoE（-/-） mice and wild-type C57BL/6 black mice（6-week old） were used in this experiment.5-Bromo-2-deoxyuridine（BrdU） was administered 24 h before the animals were sacrificed at time points of 0,2,4 or 10 weeks after hyperlipidic diet,respectively.The ascending aorta was removed for serial sectioning.The morphological changes of the aortic tissues were observed under microscope with HE staining.Some sections of the aorta were used to observe the BrdU labeling in the adventitia and intima at different time points by the method of immunohistochemistry.In addition the arterial adventitial fibroblasts derived from apoE（-/-） mice and C57BL/6 mice after hyperlipidic diet for 2 weeks were cultured.The proliferation of the cells was examined by BrdU incorporation.The cell cycle was examined by flow cytometry.RESULTS： In vivo the BrdU-labeled cells were first found in the adventitia of apoE（-/-） mice after feeding with hyperlipidic diet for 2 weeks,the time point that no visible intimal lesion formation was found,and then subsequently emerged in the intimal lesion.No BrdU labeling was observed in C57BL/6 mice at any time point.The number of BrdU labeled cells in the adventitial fibroblasts of apoE（-/-） mice was significantly higher than that in C57BL/6 mice in vitro（P0.01）.The adventitial fibroblasts of apoE（-/-） mice exhibited higher percentages of S and G2/M phases than those in C57BL/6 mice（P0.05）.CONCLUSION： The proliferation of arterial adventitial fibroblasts might participate in the early formation of atherosclerotic lesions.

关 键 词：成纤维细胞 外膜 细胞增殖 动脉粥样硬化 载脂蛋白E基因敲除小鼠 

分 类 号：R363[医药卫生—病理学]