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目的 探讨内皮黏附分子血小板内皮细胞黏附分子 1(PECAM 1)、细胞间黏附分子 3(ICAM 3)和CD4 4对淋巴管新生的作用。 方法 用狗的胸导管分离和培养淋巴管内皮细胞。标记内皮细胞的PECAM 1、ICAM 3和CD4 4 ,在荧光显微镜和共聚焦激光扫描显微镜下观察。内皮细胞用肿瘤坏死因子 (TNF α)或脂多糖 (LPS)刺激后 ,再阻断PECAM 1、ICAM 3和CD4 4 ,作细胞计数和计算迁移率。制备三维凝胶淋巴管形成模型 ,观察管状结构的形成 ,测量其长度和面积 ,并在透射电镜下观察其特征。 结果 淋巴管内皮细胞表达PECAM 1、ICAM 3和CD4 4。在对照组以及TNF α和LPS刺激组 ,分别阻断PECAM 1、ICAM 3和CD4 4后 ,内皮细胞的迁移率降低 ,管样结构的长度和面积减少。阻断PECAM 1或CD4 4后 ,细胞的增殖数目降低 ,但阻断ICAM 3后细胞的增殖数目无明显变化。在半薄和超薄切片上 ,淋巴管内皮细胞形成的管状结构具有毛细淋巴管的形态特征。 结论 体外培养的淋巴管内皮细胞表达PECAM 1、ICAM 3和CD4 4 ,这些黏附分子参与了淋巴管内皮细胞的增殖、迁移和管腔形成等淋巴管新生过程。
Objective To investigate the effects of endothelial adhesion molecule 1 (PECAM 1), intercellular adhesion molecule 3 (ICAM 3) and CD4 4 on lymphangiogenesis. Methods The lymphatic endothelial cells were isolated and cultured with dog thoracic duct. Endothelial cells were labeled with PECAM 1, ICAM 3 and CD4 4 and observed under fluorescence and confocal laser scanning microscopy. After endothelial cells were stimulated with tumor necrosis factor (TNFα) or lipopolysaccharide (LPS), PECAM 1, ICAM 3 and CD4 4 were further blocked, and cell counts and mobility were calculated. The three-dimensional gel lymphangiogenesis model was prepared. The formation of tubular structure was observed. The length and area of the tubular structure were measured. The characteristics of the structure were observed under transmission electron microscope. Results The lymphatic endothelial cells expressed PECAM 1, ICAM 3 and CD4 4. In control and TNFα and LPS groups, the migration of endothelial cells decreased and the length and area of tube-like structures decreased after blocking PECAM 1, ICAM 3 and CD4 4, respectively. After blocking PECAM 1 or CD4 4, the number of cell proliferation was decreased, but there was no significant change in the number of cells after blocking ICAM 3. In semi-thin and ultra-thin sections, the tubular structure formed by the lymphatic endothelial cells has morphological features of lymphatic vessels. Conclusion The cultured LECs express PECAM 1, ICAM 3 and CD4 4. These adhesion molecules are involved in lymphangiogenesis, such as proliferation, migration and lumen formation of lymphatic endothelial cells.