不同剂量肝素对高糖诱导后的人腹膜间皮细胞水通道蛋白1表达的影响  

Impact of different dosages' heparin on the expression of aquaporins 1 of human peritoneal mesothelial cells induced by high concentration glucose

作　　者：李鸽[1] 赵久阳[2] 任泽银[1] 肖伟[1] 桂永丰[1] 

机构地区：[1]湖南省常德市第一人民医院肾病内科,湖南常德415000 [2]大连医科大学附属第二医院肾内科,辽宁大连116000

出　　处：《中国现代医学杂志》2016年第4期18-23,共6页China Journal of Modern Medicine

摘　　要：目的研究不同剂量肝素对高糖诱导后的人腹膜间皮细胞水通道蛋白1(AQP1)m RNA和蛋白表达水平的影响。方法分离并培养人腹膜间皮细胞,并用高糖(2.5%葡萄糖溶液)诱导4、8、12和24 h;采用逆转录-聚合酶链反应(RT-PCR)检测AQP1 mRNA的表达水平,从而确定表达量达峰值的诱导时间。将不同剂量的肝素添加至人腹膜间皮细胞培养液中,并经2.5%高糖溶液诱导后,用RT-PCR及免疫组织化学法检测实验组与对照组AQP1 m RNA和蛋白表达水平。结果与对照组比较,实验组AQP1 m RNA和蛋白表达明显增加,且mRNA表达呈时间依赖性,且在高糖诱导24 h时表达量达峰值,差异有统计学意义(P<0.05);RT-PCR与免疫组织化学结果具有一致性,均提示AQP1 mRNA和蛋白表达水平在2.5%葡萄糖+1.5×10^(-2)mg/ml肝素组呈现明显上调,差异有统计学意义(P<0.05);免疫组织化学染色提示2.5%葡萄糖+1.5×10^(-2)mg/ml肝素组细胞可见胞核及胞浆有空泡样改变。结论肝素在高糖环境中诱导24 h后可显著增加人腹膜间皮细胞水通道蛋白AQP1 m RNA和蛋白的表达,且大剂量的肝素能使胞浆及胞核出现明显的空泡样改变。Objective To investigate the impact of different dosages＇ heparin on the expression of aquaporin-1（AQP1） of human peritoneal mesothelial cells（HPMCs） induced by high concentration glucose（2.5 % W/W）. Methods HPMCs were separated from patients and sub-cultured,and induced by high concentration glucose for 4,8,12,and 24h respectively. This is to determinate the time when the mRNA of AQP1 reaches the maximum level. Heparin of different dosages and 2.5% glucose were added into the cell culture medium. The expression levels of AQP1 were detected by using RT-PCR and immunohistochemistry in both control and experimental groups. Results Compared with the control group,the mRNA expression level of AQP1 was up-regulated in an time-dependant manner,with the maximum expression at 24 h induced by high concentration glucose. The protein expression level of AQP1 was also up-regulated in all experimental groups,particularly by 2.5 % glucose and 1.5×10^-2mg/ml heparin. The difference was of statistical significance（P〈0.05）. The result of RT-PCR is consistent with that of immunohistochemistry. By using immunohistochemistry,vacuolation was observed in the nucleus and cytoplasm treated by 2.5 % glu-cose and 1.5×10^-2mg/ml heparin. Conclusion The mRNA and protein expression of AQP1 can be up-regulated by 2.5 % glucose and heparin. Vacuolation can be observed in the nucleus and cytoplasm treated by glucose and heparin of high concentration.

关 键 词：肝素 人腹膜间皮细胞 水通道蛋白1 

分 类 号：R572.2[医药卫生—消化系统]