Induction of INOs expression in mouse peritoneal macrophages by Clostridium septicum

CÁCERES CLAUDIA SOLEDAD; VILLA MARÍA CECILIA; VEGA ALBA EDITH; GALLO GIOVANNA LUCRECIA; ORTEGA NICOLAS; CORTIÑAS TERESA INÉS; STEFANINI ANA MARÍA

Mendoza

Congreso; XXIV Reunión Científica Anual de La Sociedad de Biología de Cuyo.; 2008

Sociedad de Biología de Cuyo

Clostridium septicum is the causative agent of traumatic gangrene in most animals and endogen myonecrosis in man. Without a treatment all the infections are of rapid establishment and inevitably fatal. Previous results showed the capacity of Clostridium septicum to induce necrosis or apoptosis on mouse peritoneal macrophage (Mq) depending of the infective doses. Both are considered mechanisms for evasion of innate immune system as they achieve the destruction of key immune system cells. The aim of this study was to analyze the induced nitric oxide sintetase (iNOS) expression of Mq induced by C. septicum infection. Cells of C. septicum ATCC 12464 collected at logarithmic phase of grow in a clostridial medium under anaerobic conditions were used to infect Mq. The expression of iNOs gene and B actin as housekeeping gene were analyzed with the multiplicity of infection (moi) 1:10 y 1:100 at 4 and 24 h. For RNA extraction, 106 cells/well were treated with TRIzol reagent. cDNA was performed with random hexamer and 200 U Moloney murine leukaemia virus reverse transcriptase. The results showed increased iNOS expression at moi 1:100 at 4 h and a lost of it at 24 h meanwhile at moi 1:10 a low expression of iNOS was sustained both at 4 and 24 h. The expression of iNOs confirmed the previous hypothesis on the effect of nitric oxide (NO) inducing apoptosis or necrosis depending of the levels of NO produced by the Mq.