کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2549132 1124501 2011 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
An ex vivo standardized assay to measure human platelet cGMP
موضوعات مرتبط
علوم پزشکی و سلامت داروسازی، سم شناسی و علوم دارویی داروشناسی
پیش نمایش صفحه اول مقاله
An ex vivo standardized assay to measure human platelet cGMP
چکیده انگلیسی

IntroductionNitric oxide (NO) acts a pleiotropic biomodulator in several systems, including the cardiovascular, nervous and immune systems. The intracellular levels of cyclic guanylate monophosphate (cGMP) can be increased by NO or by inhibiting the breakdown to 5′cGMP operated by the cyclic nucleotide phosphodiesterases (PDEs). Platelets are anuclear circulating cells that are rich in both soluble guanylyl cyclase and PDEs. The purpose of this study was to standardize cGMP determination in human platelets.MethodsFresh blood samples were obtained from a group of healthy volunteers in order to obtain washed platelets. Platelet (3 × 104/μl or 5 × 105/μl) cGMP levels were measured in basal and stimulated conditions. To stimulate platelets two different NO-donors were used: sodium nitroprusside (SNP; 10, 100, 1000 μM) or diethylamine NONOate (DEA-NONOate; 1, 10, 100 μM). Different times of incubation were also studied (5, 15 and 30 min). As positive control has been used ODQ a well known inhibitor of guanylyl cyclase. Platelet cGMP accumulation was measured by using a standard ELISA kit using different sample dilutions.ResultsThe optimal stimulus was DEA-NONOate, the optimal washed platelet concentration was 5 × 105/μl, incubation time was 30 min and dilution to be used was 1:2.DiscussionPlatelets represent a valuable marker to investigate the effect of drugs interfering with the cGMP cascade. This standardized assay allows to measure ex vivo the inhibitory (PDE inhibitors) or stimulatory effect (NO donors) of drugs given in vivo to humans.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmacological and Toxicological Methods - Volume 64, Issue 2, September–October 2011, Pages 164–167
نویسندگان
, , , , , , ,