摘要目的 观察CD44抗体对视网膜Müller细胞降解透明质酸(HA)功能的影响.方法 培养的猪后极部视网膜Müller细胞,取第2代细胞用于实验.细胞分为1、2、3组,每组含12×103个Müller细胞.其中1组培养液中未加入其他试剂;2组培养液中加入0.01 mg/ml HA;3组培养液中加入0.01 mg/ml HA和10μg/ml抗CD44抗体.2、3组细胞和HA、单克隆抗CD44抗体预培养,收集1、2、3组上清液,行HA-底物胶电泳法和类酶链免疫吸附测定(ELISA)法检测.结果 HA-底物胶电泳法分析结果显示,1组表现为蓝色背景下的白色细淡的双条带;2组双条带明显增厚,合并成较粗和明亮的脱色条块;3组脱色条块回复为细淡的双条带.类ELISA法检测结果显示,1、2、3组吸光度[A,旧称光密度(OD)]值分别为0.310±0.025、0.093±0.051、0.025±0.069.2组A值较1组A值明显降低,与1组A值比较,差异有统计学意义(t=28.1,P＜0.01);3组A值与1组A值比较,差异无统计学意义(t=4.92,P＞0.05),与2组A值比较,差异有统计学意义(t=26.9,P＜0.01).结论 Müller细胞与HA的相互作用可加强细胞降解HA的功能,CD44抗体可降低这种加强作用.

abstractsObjective To observe the effect of CD44 antibody on the hyaluronic acid (HA)generation) were cultured in three different medium: without HA (group 1),0. 01 mg/ml HA (group 2),10 μg/ml HA and CD44 antibody (group 3). The cells in the group 2 and 3 were pre-cultured with HA and CD44 antibody, and the supernatant was collected. HA-substrate gel electrophoresis was performed for HA degradation, while ELISA-like method was performed for HA-binding protein. Results HA-substrate gel electrophoresis showed white light double-band on blue background in groups 1 and 3, thicker double-band or bright de-colored blocks in group 2. ELISA-like method showed that the absorbance (A) value of groups 1,2 and 3 were 0.310 ± 0.025, 0.093 ± 0.051, 0.025 ± 0.069 respectively. The A value of group 2 was obviously lower than that of group 1 (t=28.1, P＜0.01); the A value of group 3 was significantly higher than that of group 2 (t=26.9, P＜0.01), but was the same as group 1 (t=4.92, P＞0.05). Conclusion CD44 antibody can inhibit the interaction between Miller cells and HA, and thus reduce the HA degradation.