RESUMO
Pregnane X receptor (PXR) is a member of nuclear receptor subfamily 1 (NR1I2) that is a transcriptional regulator of several metabolic enzymes involved in clopidogrel metabolism. In this study we identified and evaluated the contributions of single nucleotide polymorphisms (SNPs) in NR1I2 and cytochrome P450 (CYP) 2C19 alleles to clopidogrel resistance (CR) and long-term clinical outcomes in acute ischemic stroke (IS) patients. A total of 634 patients with acute IS were recruited, who received antiplatelet medication (clopidogrel or aspirin) every day and completed a 1-year follow-up. The selected SNPs were genotyped, and platelet function was measured. Modified Rankin Scale (mRS) scores and main adverse cardiovascular and cerebrovascular events (MACCE) were noted to assess the prognosis. We showed that SNPs NR1I2 rs13059232 and CYP2C19 alleles (2*/3*) were related to CR. SNP NR1I2 (rs13059232) was identified as an independent risk factor for the long-term clinical outcomes in the clopidogrel cohorts (P < 0.001), but similar results were not observed in a matched aspirin cohort (P > 0.05). Our results suggest that NR1I2 variant (rs13059232) could serve as biomarker for clopidogrel therapy and individualized antiplatelet medications in the treatment of acute IS patients.
Assuntos
Infarto Encefálico/tratamento farmacológico , Infarto Encefálico/genética , Clopidogrel/uso terapêutico , Inibidores da Agregação Plaquetária/uso terapêutico , Polimorfismo de Nucleotídeo Único , Receptor de Pregnano X/genética , Doença Aguda , Idoso , Idoso de 80 Anos ou mais , Aspirina/uso terapêutico , Infarto Encefálico/diagnóstico , Estudos de Coortes , Citocromo P-450 CYP2C19/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Agregação Plaquetária/genética , Prognóstico , Fatores de RiscoRESUMO
Trimethylamine-N-oxide (TMAO) is derived from the gut microbiome and tissues metabolism of dietary choline and betaine. These molecules are closely related to the development of cardiovascular and cerebrovascular diseases. A rapid, sensitive and accurate method has been developed and validated for the simultaneous determination of trimethylamine N-oxide (TMAO), choline and betaine in human plasma using d9-trimethylamine N-oxide (TMAO), d9-choline, d9-betaine as the internal standard (IS). After methanol precipitation with 10⯵L plasma samples, the analytes were extracted and then separated on Amide column (2.1â¯×â¯100â¯mm, 1.7⯵m, waters) with an isocratic elution program consisting of acetonitrile-water (containing 10â¯mM ammonium formate pHâ¯=â¯3.0) at a flow of 400⯵L/min. The detection was achieved under the selected reaction monitoring (SRM) scan using positive electrospray ionization (ESI+) in 3â¯min. The mass transitions monitored were as follows: m/z 76.3â¯ââ¯58.4 for TMAO, m/z 104.2â¯ââ¯60.3 for choline, m/z 118.1â¯ââ¯58.3 for betaine, m/z 85.1â¯ââ¯66.3 for d9-TMAO, m/z 113.2â¯ââ¯69.3 for d9-choline, and m/z 127.1â¯ââ¯67.2 for d9-betaine, respectively. The method has been fully validated for specificity, lower limit of quantification, linearity, stability, intra- and inter-day accuracy and precision. This assay combines simple sample processing with a short run time and small plasma volumes, making it well suited for high-throughput routine clinical or research purposes. The newly developed method was successfully applied to the patients (nâ¯=â¯220) suffered from acute stroke, and the concentration of choline was firstly found to be closely related with the prognosis of these patients.