【摘要】 目的探讨一氧化氮(NO)和溶血卵磷脂对人脐静脉内皮细胞胞外5′-外核苷酸酶(CD73)的调节作用。方法将已融合人脐静脉内皮细胞的细胞培养皿分为8组(n=15),即对照组:无干预;LPC组:培养皿中加入LPC 10μmol/L;AOPCP组:加入胞外5′-外核苷酸酶抑制剂α,β-甲基腺苷-5-二磷酸50μmol/L。另外5个均为NO组,分别加入NO供体亚硝基铁氢化钠25,100,300,500或1000μmol/L。每组均于试验开始时加入乙烯-单磷酸腺苷(eAMP,5μmol/L)。在试验第15、30、45 min时以高性能液相色谱仪测定各培养皿中乙烯-腺苷含量。结果在上述三个时间点LPC组乙烯-腺苷生成较对照组显著增高(P<0.05),而各不同浓度NO组乙烯-腺苷生成与对照组无显著差别(P>0.05),AOPCP组乙烯-腺苷生成较对照组显著减少(P<0.01)。结论LPC可兴奋有脐静脉内皮细胞外5′-核苷酸酶,增加腺苷生产率;而NO对离体人内皮细胞外5′-核苷酸活性无明显影响。更多还原

【Abstract】 Objective To evaluate the effect of nitric oxide(NO) and phosphatide on ecto-5′-nucleotidase activity in Human umbilical endothelial cells(HUVEC).Methods Experiments were conducted in HUVEC grown on dishes which were divided into 8 groups: Control group in which only eAMP(5 μM) was added;LPC group in which HUVEC were incubated with LPC(10 μM) before eAMP was added;AOPCP group in which cells were incubated with AOPCP(10 μM) before eAMP was added.Other five groups are NO groups in which 25,100,300,500 or 1000 μmol/L SNAP(NO doner) was added respectively.Etheno-adenosine production was detected at 15th,30th,45th min with high performance liquid chromatography(HPLC) respectively.Results Comparing to the control group LPC significantly increased etheno-adenosine production at three time points respectively(P<0.05,n=15);However,no significant difference were found among control and all these five NO groups(P>0.05,n=15).AOPCP significantly decreased the etheno-adenosine production compared to the control at three time points respectively(P<0.01,n=15).Conclusions Ecto-5′-nucleotidase can be up-regulated within minutes following exposure of HUVEC to LPC;However,NO has no effect on ecto-5′-nucleotidase in human endothelial cells.更多还原