Glucocorticoids as regulators of gene expression on human macrophages

DIAZ LUDOVICO IVO; ESTEVE RAFOLS, MONTSERRAT; LEDDA, ANGELO; GRASA, MARÍA DEL MAR; TOLEDO, J. D.; GARDA, H. A.; GONZALEZ MARINA CECILIA

Rosario

Congreso; 49th Reunión anual. Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2014

SAIB

Human monocytes THP1 cells were transformed into macrophages by adding PMA in the culture medium. Macrophages THP1 cells were treated with Ox-LDL/cortisol or Ox-LDL/cortisone or Ox-LDL/cortisone/BVT.2733 inhibitor for 24 h.  Isolated LDL fraction from human plasma was peroxidized in vitro with Cu++ to transform LDL into Ox-LDL. We evaluated the expression of genes involved in the pro inflammatory (TNFα, FAT/CD36, IL6) and anti inflammatory processes (11ßHSD1, rIL10, MMR). The 11ßHSD1 expression is involved in the reduction of cortisone to cortisol (glucocorticoid active form) and the 11ßHSD2 turns cortisol into cortisone. The mRNA level of different genes was measured by quantitative real-time PCR. Results showed a decrease of pro-inflammatory marker in cells treated with increasing cortisol doses. Also, the treatment with high cortisone concentration (1000nM) showed decrease of pro-inflammatory gene expression. However, cells treated with increasing cortisone concentration and 11ßHSD1 inhibitor (preventing conversion of cortisone into cortisol) did not show pro-inflammatory gene expression decrease. Enhancing cortisol concentration promotes a decrease of 11ßHSD1 and an increase of 11ßHSD2 gene expression. In conclusion, our results showed that only cortisol contributes to an anti-inflammatory response.