Fig. 7

The CYP1A1–12(S)-HETE-JNK-AP-1 signalling axis is activated in septic patients. a, b Human peripheral monocytes were isolated from healthy donors and treated with heat-killed E. coli (MOIs = 10) at the indicated times. Supernatants were collected for analysis of TNF-α, IL-6 and 12(S)-HETE levels using ELISA. Cell lysates were used for JNK/AP-1 phosphorylation assessment. Mean of three independent experiments. Results were compared by one-way ANOVA. c CYP1A1 mRNA levels in peripheral monocytes from the studied subjects were measured by qRT-PCR. TNF-α, IL-6 and 12(S)-HETE levels in plasma were analysed by ELISA (n = 30 for healthy individuals, n = 30 for sepsis patients). Group means were compared by Student’s t test. Correlation of SOFA scores with CYP1A1 (r = 0.65, P < 0.05) and 12(S)-HETE (r = 0.38, P < 0.05) in sepsis patients. d Peripheral monocytes were isolated from 10 septic patients and 10 healthy controls and cultured for 2 h, respectively. Supernatants were collected for analysis of TNF-α, IL-6 and 12(S)-HETE levels using ELISA. Group means were compared by Student’s t test. e Peripheral monocytes were isolated from 6 septic patients and 6 healthy controls and coaxed into macrophages by PMA for 48 h. Supernatants were collected for analysis of TNF-α, IL-6 and 12(S)-HETE levels using ELISA. CYP1A1 mRNA levels in monocytes-derived macrophages were measured by qRT-PCR. Group means were compared by Student’s t test. f CYP1A1 protein, JNK phosphorylation and AP-1 phosphorylation levels in peripheral monocytes from healthy individuals and septic patients were assessed using LSCM. Bar, CYP1A1: 5 μm, JNK/AP-1: 10 μm. Data shown as mean ± SEM of triplicate experiments. * p < 0.05