Background The gout is an inflammatory multifactorial disorder where the diet, age, sex, absorption regulation of uric acid in kidney and genetic, contribute to the onset of the disease. The balance of uric acid concentration not only depends on metabolism of purines but also on the clearance of uric acid, in which many proteins participate in the reabsorption and transport of urate. Is unknown if the peripheral blood leukocyte cells can change their expression and regulation mechanism of the urate transporters by the presence of uric acid in gout patients [1–4].
Objectives Analyze changes in gene expression and the methylation pattern of the TLR2, TLR4, SLC2A9, SLC22A12, SLC22A3 and ABCG2 in neutrophils and peripheral blood monocytes from patients with gout and controls
Methods The isolation of peripheral blood neutrophils and monocytes cells was performed by negative immunomagnetic selection (MACxpress kit, EUA). By flow citometry were analyzed the previously separate cell populations, mononuclear (MN), polymorphonuclear (PMN) cells and neutrophils (N) (CD15, CD16, CD14). The DNA and RNA extraction was realized with a without columns kit and with Trizol technique. The gene expression analysis will be performed from total RNA by RT-PCR kit (Promega). Methylation analysis will be carried out the bisulfite conversion (ABCAM) from total DNA and HRM-PCR. For both studies, gene expression and methylation analysis are designed specific primers
Results The biggest difference between asymptomatic gout patients (n=12) and controls (n=12), in the biochemical parameters (Table 1), is in the higher levels of uric acid and triglycerides that the patient presents. Actually, we've already evaluated the genetic expression of TLR1, TLR2, TLR4 and IL1b in mononuclear cells of 5 asymptomatic gout patients and 5 controls (Plot 1). Interestingly, IL1b is UP-regulated in sample group bye a mean factor of 16.350 and TLR2 is UP-regulated in sample group in comparison to control group by a mean factor of 3.686.
Conclusions The asymptomatic patients with a higher mean of uric acid (8mg/dL) and tryglicerides (145.60 mg/dL) had a higher expression of IL1b and TLR2 compared to controls.
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Disclosure of Interest None declared