Article Text

PDF
AB0861 Expression control by methylation of the TLR1, TLR2, TLR4, IL1B, ALPK1 SLC2A9 and SLC22A12 genes in monocytes of patients with gout
  1. N Díaz Paniagua1,
  2. EG Tranquilino Batres1,
  3. AP Lόpez Flores2,
  4. A Lozano Cardenas2,
  5. E Vallarino Reyes2,
  6. AL Alvarez Grijalva2,
  7. L Sanchez Chapul1,
  8. C Díaz Hernández3,
  9. L Ríos Ventura3,
  10. A Lopez Macay1
  1. 1Enfermedades Neuromusculares, Instituto Nacional de Rehabilitacion
  2. 2Ciencias de la vida, Instituto de Estudios Superiores Monterrey
  3. 3LUMA, Instituto Nacional de Rehabilitacion, Mexico City, Mexico

Abstract

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.

Table 1.

Characteristics of patients and controls

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.

References

  1. Neogi T., et al. Gout Classification. Criteria An American College of Rheumatology/European League Against Rheumatism. Arthritis & Rheumatology. 2015.

  2. Busso N., et al. Review Mechanisms of inflammation in gout. Arthritis Research & Therapy 2010.

  3. Mitroulis I., et al. Review, Neutrophils, IL-1β, and gout: is there a link. Semin Immunopathology. 2013.

  4. Partial HPRT. Deficiency with a Novel Mutation of the HPRT Gene in Combination with Four Previously Reported Variants Associated with Hyperuricemia. International Medicine. 2014.

References

Disclosure of Interest None declared

Statistics from Altmetric.com

Request permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.