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高葡萄糖水平对人血小板NTPD酶和5'-核苷酸酶活性的影响。

Effect of high glucose levels in human platelet NTPDase and 5'-nucleotidase activities.

作者信息

Lunkes Gilberto Inácio, Lunkes Daniéle Sausen, Leal Daniela, Araújo Maria do Carmo, Corrêa Maísa, Becker Lara, Rosa Cintia Saydelles da, Morsch Vera Maria, Schetinger Maria Rosa Chitolina

机构信息

Departamento de Química, Centro de Ciências Naturais e Exatas, Programa de Pós-Graduação em Bioquímica Toxicológica, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil.

出版信息

Diabetes Res Clin Pract. 2008 Sep;81(3):351-7. doi: 10.1016/j.diabres.2008.06.001. Epub 2008 Jul 21.

DOI:10.1016/j.diabres.2008.06.001
PMID:18644642
Abstract

OBJECTIVES

The objective of this work was to evaluate the effect of different glucose levels on the ATP, ADP and AMP hydrolysis in the platelets of diabetic, hypertensive and diabetic/hypertensive participants.

METHODS

The activities of the enzymes NTPDase (ATP and ADP hydrolysis) and 5'-nucleotidase (AMP hydrolysis), and CD39 expression were analyzed in human blood platelets of diabetic (DM-2), hypertensive (HT) and diabetic/hypertensive (DM-2/HT) patients. To evaluate the interference of glucose and fructose in NTPDase and 5'-nucleotidase activities, experiments were performed with glucose, fructose and mannitol concentrations ranging from 5 to 30 mM in platelet-rich plasma (PRP). Pre-incubation times of 10, 120 min and 24h were used.

RESULTS

NTPDase and 5'-nucleotidase activities increased with increasing glucose and fructose concentrations (P<0.001) and the different times of pre-incubation did not interfere in ectonucleotidases activities (P>0.5). NTPDase and 5'-nucleotidase activities demonstrated a positive correlation between serum glucose levels and ATP and ADP hydrolysis in DM-2 and DM-2/HT patients. CD39 expression demonstrated that DM-2, HT and DM-2/HT groups presented a significant increase when compared to the control group (P<0.004).

CONCLUSION

The hydrolysis of adenine nucleotides is enhanced in platelets of patients with diabetes and hypertension. We observed that an increasing glucose concentration had a direct effect on ATP, ADP and AMP hydrolysis. Furthermore, CD39 expression was enhanced in all patients groups, indicating that these enzyme activities are related with diabetes and hypertension.

摘要

目的

本研究旨在评估不同葡萄糖水平对糖尿病、高血压及糖尿病合并高血压患者血小板中三磷酸腺苷(ATP)、二磷酸腺苷(ADP)和一磷酸腺苷(AMP)水解的影响。

方法

对糖尿病(2型糖尿病,DM-2)、高血压(HT)和糖尿病合并高血压(DM-2/HT)患者的人血小板进行分析,检测三磷酸核苷二磷酸酶(NTPDase,ATP和ADP水解)和5'-核苷酸酶(AMP水解)的活性以及CD39的表达。为评估葡萄糖和果糖对NTPDase和5'-核苷酸酶活性的干扰,在富含血小板血浆(PRP)中用5至30 mM的葡萄糖、果糖和甘露醇浓度进行实验。预孵育时间分别为10、120分钟和24小时。

结果

NTPDase和5'-核苷酸酶活性随葡萄糖和果糖浓度的增加而增加(P<0.001),不同的预孵育时间对胞外核苷酸酶活性无干扰(P>0.5)。在DM-2和DM-2/HT患者中,NTPDase和5'-核苷酸酶活性与血清葡萄糖水平以及ATP和ADP水解呈正相关。CD39表达显示,与对照组相比,DM-2、HT和DM-2/HT组均有显著增加(P<0.004)。

结论

糖尿病和高血压患者血小板中腺嘌呤核苷酸的水解增强。我们观察到葡萄糖浓度的增加对ATP、ADP和AMP水解有直接影响。此外,所有患者组的CD39表达均增强,表明这些酶活性与糖尿病和高血压有关。

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