摘要
目的通过观察一氧化氮对腹膜间皮细胞醛糖还原酶(AR)活性的影响,探讨一氧化氮对腹膜间皮细胞醛糖还原酶的调节作用。方法采用人的网膜作为细胞来源,胰蛋白酶-EDTA消化法进行人腹膜间皮细胞(HPMC)的分离、培养,间接免疫荧光法对第二代细胞进行鉴定。第2～3代细胞用于实验。以硝普纳为一氧化氮供体,硝普纳(0.5mmol/L、1mmol/L、2mmol/L)+1.5%葡萄糖培养液为实验组,以无一氧化氮1.5%葡萄糖培养液为对照组,观察剂量及时间对腹膜间皮细胞醛糖还原酶活性的影响。应用紫外分光光度计记录NADPH在340nm处吸光值的下降表示醛糖还原酶活性。结果高糖状态下各浓度一氧化氮组醛糖还原酶活性均高于对照组,且醛糖还原酶活性随一氧化氮浓度的增加而增加;一氧化氮+葡萄糖组醛糖还原酶活性随着时间的延长而逐渐增加。各时点一氧化氮+葡萄糖组醛糖还原酶活性均高于葡萄糖组。结论一氧化氮以时间及剂量依赖的方式使腹膜间皮细胞醛糖还原酶活性增加。
Objective To observe the effect of nitric oxide on activity of aldose reductase in peritoneal mesothelial cells and explore its regulation. Methods Mesothelial cells were isolated from the omental tissue of consenting patients by typsin-EDTA disaggregation and indirect immunofluorescence was used for identification. HPMCs were treated with D-glucose（ 1.5% ） in presence of increasing NO concentration （0.5mmol/L, 1.0mmol/L,2.0mmol/L）. HPMCs of control group was just incubated with 1.5% D-glucose. Aldose reductase activity was measured on spectrophotometer by monitoring the decrease in absorbance of NADPH at 340nm. Results A dose-dependent increase in AR activity was observed with increasing NO concentration. Incubation of HPMC with glucose in the presence of NO resulted in a time-dependent increase in AR activity. In contrast in the presence of NO glucose-induced increase of AR activity was greater than glucose alone. Conclusion Nitric oxide can up-regulate AR activity in a dose and time dependent way.
出处
《中国现代医生》
2008年第5期8-10,共3页
China Modern Doctor
关键词
一氧化氮
腹膜间皮细胞
醛糖还原酶
多元醇通路
腹膜透析
Nitric oxide
Peritoneal mesothelial cells
Aldose reductase
Polyol Pathway
Peritoneal dialysis
