Figure 5

Inhalation of CO gas and administration of CORM-2 and hemin increase IRG1 and A20 expression via HO-1 expression and decrease inflammation in a sepsis mouse model in vivo. (a–c) Wild-type 7-week-old male C57BL/6 mice were inhaled with CO gas (250 ppm) for 6 days (4 h daily basis) and some of them were administrated with ZnPP (5 mg/kg; i.p.). After 6 days, the mice were administrated with LPS (12.5 mg/kg; i.p.) for 16 h. (a) Blood serum was analyzed for the TNF-α level by ELISA. (b) Liver tissues were analyzed for the levels of TNF-α by real-time RT-PCR. (c) Liver tissues were analyzed for the levels of IRG1, A20 and HO-1 mRNA by real-time RT-PCR. (d–f) Wild-type 7-week-old male C57BL/6 mice were pre-treated with CORM-2 (30 mg/kg; i.p.), hemin (10 mg/kg; i.p.) and ZnPP (5 mg/kg; i.p.) for 2 h and, then, the mice were administrated with LPS (12.5 mg/kg; i.p.) for 16 h. (d) Blood serum was analyzed for the TNF-α level by ELISA. (e) Liver tissues were analyzed for the levels of TNF-α by real-time RT-PCR. (f) Liver tissues were analyzed for the levels of IRG1, A20 and HO-1 mRNA by real-time RT-PCR. Data represents mean±s.e.m., * P<0.05 and ** P<0.001 as compared with control; and ^# P<0.05 and ^## P<0.001 as compared with the cells exposed to LPS+CO/CORM-2/hemin (in case TNF-α, ^## P<0.001 as compared with the cells exposed to only LPS).