表皮生长因子和维生素A对高浓度葡萄糖抑制腹膜间皮细胞增殖和分化的影响

Effects of epidermal growth factor and vitamin A on proliferation and differentiation of human peritoneal mesothelial cells inhibited by high concentration glucose

目的 研究表皮生长因子（ Ｅ Ｇ Ｆ） 和维生素 Ａ（ Ｖｉｔ Ａ） 对高浓度葡萄糖抑制腹膜间皮细胞增殖和分化的影响，以探讨 Ｅ Ｇ Ｆ和 Ｖｉｔ Ａ 在腹膜修复中的作用。方法 采用胰蛋白酶消化法，从人腹膜组织中分离间皮细胞，并建立体外人腹膜间皮细胞（ Ｈ Ｐ Ｍ Ｃ） 培养模型；以３ Ｈ胸腺嘧啶核苷（３ Ｈ Ｔｄ Ｒ） 掺入法测定细胞内 Ｄ Ｎ Ａ 含量示细胞增殖程度；以免疫组织化学染色法，用图像分析仪测细胞表面角蛋白和波形蛋白（ｖｉｍｅｎｔｉｎ） 表达量示细胞分化程度。结果 随着葡萄糖浓度增高和培养时间的延长， Ｈ Ｐ Ｍ Ｃ３ Ｈ Ｔｄ Ｒ 掺入降低； Ｅ Ｇ Ｆ 和 Ｖｉｔ Ａ 可增加 Ｈ Ｐ Ｍ Ｃ 对３ Ｈ Ｔｄ Ｒ 的掺入，但两者ｃｐｍ 值比较无显著性差异（ Ｐ＞ ００５） ；在含高浓度葡萄糖的培养液中分别加入 Ｅ Ｇ Ｆ或 Ｖｉｔ Ａ 后， Ｅ Ｇ Ｆ 可明显促进 Ｈ Ｐ Ｍ Ｃ 的增殖，其３ Ｈ Ｔｄ Ｒ 掺入量明显增加，与 Ｖｉｔ Ａ 组比较有显著性差异（ Ｐ＜ ００５） 。 Ｅ Ｇ Ｆ 和 Ｖｉｔ Ａ 不影响细胞表面角蛋白和波形蛋白的表达。结论 Ｅ Ｇ Ｆ和 Ｖｉｔ Ａ 对腹膜间皮细胞的分化未显示出明显作用，但两者均可刺激细胞的增殖，特别是 Ｅ Ｇ

Objective To study the effects of EGF and VitA on the proliferation and differentiation of human peritoneal mesothelial cells(HPMC) inhibited by high concentration glucose and investigate their roles in the repair of peritoneal mesothelium. Methods HPMC was separated with trypsin and the synthesis of DNA was measured by 3H TdR incorporation.The expression of keratin and vimentin on the surface of HPMC were measured by immunohistochemistry staining. Results EGF and VitA didn't effect the expression of keratin and vimentin on the surface of HPMC. 3H TdR incorporation of HPMC cultured in the medium containing high concentration glucose was decreased significantly than in the RPMⅠ 1640 alone( P <0 05). EGF and VitA both increased the 3H TdR incorporation,showing no significant difference.But 3H TdR incorporation of HPMC cultured in the medium with high concentration glucose and EGF was increased significantly( P <0 05)than in the medium with high concentration glucose and VitA. Conclusion EGF and Vit A do not effect the differentiation of HPMC,but can ameliorate the inhibitive effect on proliferation of HPMC by high concentration glucose.It may play an important role in the repair of damaged mesothelium and restoration of its function.