历志, 魏旭峰, 顾春虎, 康晓军, 王春梅, 韩骅, 易定华. 正常小鼠主动脉瓣膜的组织形态学研究[J]. 心脏杂志, 2010, 22(5): 677-683.
    引用本文: 历志, 魏旭峰, 顾春虎, 康晓军, 王春梅, 韩骅, 易定华. 正常小鼠主动脉瓣膜的组织形态学研究[J]. 心脏杂志, 2010, 22(5): 677-683.
    Histological and morphometric analyses of mouse heart valve aorta structure[J]. Chinese Heart Journal, 2010, 22(5): 677-683.
    Citation: Histological and morphometric analyses of mouse heart valve aorta structure[J]. Chinese Heart Journal, 2010, 22(5): 677-683.

    正常小鼠主动脉瓣膜的组织形态学研究

    Histological and morphometric analyses of mouse heart valve aorta structure

    • 摘要: 目的: 建立转基因小鼠主动脉瓣疾病模型的评估标准。方法: 采用3月龄C57成年小鼠20只,分离小鼠心脏称重后固定,经HE染色、免疫荧光、激光共聚焦和Massoon染色后光镜观察及电镜观察,分别对瓣膜的细胞数量、分布、胶原含量及瓣膜超微结构进行组织形态学分析。结果: 正常小鼠主动脉瓣的面积为(0.1978±0.003) mm2,周长为(9.18±0.06)×102 μm,平均细胞数为(9.75±0.04)×102个/mm2,其中内皮细胞数为(3.23±0.10)×102个/mm2,间质细胞数为(5.05±0.07)×102个/mm2,胶原含量为(23.77±8.38)%。此外,通过激光共聚焦显微镜观察还发现,成体内皮下层存在一些共表达CD31及α-SMA的细胞,表明成体瓣膜仍保留具有上皮-间质转化潜能的细胞。电镜观察发现,瓣膜的心室面内皮与主动脉面内皮在形态上存在有很大的差异:心室面内皮呈扁平形与血流方向一致,主动脉面内皮呈方形与血流方向垂直。间质中成纤维细胞处于静止状态,其细胞胞体较小,呈长梭形,粗面内质网和高尔基复合体均不发达。细胞外基质胶原的含量丰富,主要集中于流出道。结论: 本实验首次详细观测了小鼠主动脉瓣形态学特征,建立了正常小鼠主动脉瓣组织形态学研究的标准,为今后小鼠主动脉瓣疾病模型的研究提供重要的参考依据。

       

      Abstract: AIM: To provide standard morphometric data of normal mouse aorta valve structure for the evaluation of transgenic mouse models of aortic valve diseases. METHODS: Adult mouse (postnatal 3 months) hearts were harvested and histological and valve morphometric analyses of valve cell population, distribution and collagen content as well as valve microstructure were conducted by light microscope (HE stain, immunofluorescence, Laser scanning confocal microscope and Masson’s trichrome stain) and transmission electron microscope. RESULTS: The aortic valve area of normal mice was (0.1978±0.003) mm2 with a perimeter of (9.18±0.06)×102 μm, average cell number (9.75±0.04)×102/mm2, endothelial cell number (3.23±0.10)×102/mm2, intersititial cell number (5.05±0.07)×102/mm2, and collagen content (21.62±9.33)%. Confocal laser scanning displayed some co-expressed CD31 and α-SMA cells under endothelium, indicating the existence of epidermal-mesenchymal transformation potential cells in adult aortic valves. Electron microscopy showed some morphological differences between the ventricular face of the valve and the aortic face of the valve endothelium. The ventricular face of the valve endothelium was flat, whereas the aortic face of the valve endothelium was square. Interstitial fibroblasts were in a quiescent state in which the cell bodies were short and long spindle-shaped, and endoplasmic reticulum and Golgi complexes were undeveloped. Extracellular matrix was collagen-rich, which was mainly concentrated in the outflow tract. CONCLUSION: Histological and morphometric data are provided for normal mouse aortic valve structure and are useful as reference standards for future studies of mouse models of progressive aortic valve diseases.

       

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