Inhibitive effect of exogenous carbon monoxide-releasing molecules 2 on the activation of Janus kinase/signal transducer and activator of transcription pathway in sepsis / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the inhibitive effect of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on the activation of Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway in sepsis.</p><p><b>METHODS</b>RAW264.7 cells were divided into normal control group, LPS group (10 mg/mL LPS, the same concentration below), LPS + inactive CORM-2 (iCORM-2) group, LPS + 50 mmol/L CORM-2 group, and LPS + 100 mmol/L CORM-2 group. TNF-alpha level in the supernatant was determined with ELISA, and the phosphorylation levels of JAK1 and JAK3 were determined with Western blot. Thirty-five male BALB/c mice were divided into normal control group, cecal ligation and puncture (CLP) group, CLP + iCORM-2 (8.0 mg/kg) group and CLP + CORM-2 group (8.0 mg/kg) according to the random number table. Mice in CLP + CORM-2 group were treated the same as mice in CLP group except for administration of CORM-2 after CLP. The plasma levels of TNF-alpha, IL-1beta, and the phosphorylation levels of JAK1, JAK3 in liver tissue were determined with ELISA 24 hours post CLP. Data were processed with t test.</p><p><b>RESULTS</b>Compared with that of normal control group [(1.9 +/- 0.3) pg/mL], the TNF-alpha level [(8.2 +/- 2.7) pg/mL, t = 2.844, P < 0.01] and phosphorylation levels of JAK1, JAK3 in LPS group increased significantly; while TNF-alpha levels in LPS + 50 mmol/L CORM-2 and LPS + 100 mmol/L CORM-2 groups decreased obviously as compared with that of LPS group [(5.7 +/- 1.4), (3.2 +/- 0.9) pg/mL, with t value respectively 2.104 and 2.363, P values all below 0.05], and it was the same with phosphorylation levels of JAK1, JAK3 in a dose-dependent manner. Compared with those of normal control group, plasma levels of TNF-alpha and IL-1beta and phosphorylation levels of JAK1, JAK3 in liver tissue significantly increased in CLP group (with t value respectively 2.916 and 2.796, and P values all below 0.05); while plasma levels of TNF-alpha and IL-1beta and the phosphorylation levels of JAK1, JAK3 in liver tissue decreased significantly in CLP + CORM-2 group (with t value respectively 2.115 and 2.398, and P values all below 0.05).</p><p><b>CONCLUSIONS</b>Exogenous CORM-2 can obviously inhibit the phosphorylation of JAKs molecules and then inhibit the activation of JAK/STAT signal pathway in sepsis, and decrease the expression of downstream cytokines to effectively prevent cascade reaction in the inflammatory response after severe infection.</p>

Inhibitive effect of exogenous carbon monoxide-releasing molecules 2 on tissue factor expression in sepsis / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the inhibitive effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on expression of tissue factor (TF) in sepsis.</p><p><b>METHODS</b>Human umbilical vein endothelial cells (HUVEC) were cultured with trypsin digestion method, which were divided into NC group (with normal treatment), LPS group (with culture of 10 microg/mL LPS), LD group (with 10 microg/mL LPS and DMSO in co-culture), LC1 group (with 10 microg/mL LPS and 10 micromol/L CORM-2 in co-culture), LC2 group (with 10 microg/mL LPS and 50 micromol/L CORM-2 in co-culture), LC3 group (with 10 microg/mL LPS and 100 micromol/L CORM-2 in co-culture). After culture for 4 hours, TF activity, TF protein expression, nuclear factor-kappaB (NF-kappaB) activity were examined. Forty-five C57 BL/6 male mice were randomly divided into NC (without treatment, n = 5), CLP (n = 5) and CLP + CORM-2 (with treatment of 8 mg/kg CROM-2 after CLP, n = 5) groups. The serum samples in CLP, CLP + CORM-2 groups were collected at 2, 6, 12 and 24 post operation hour ( POH, 5 mice at each time point) for determination of TF and TFPI levels,which were also examined in NC group.</p><p><b>RESULTS</b>Compared with those of NC group, TF activity increased (P < 0.01) , TF protein expression and NF-KB activity also increased in LPS group. Compared with those of LPS group, above indices were decreased in LC1, LC2, LC3 groups. The serum level of TF in CLP group at 6 POH was higher than that of NC group (80.0 +/- 11.9 pg/mL vs 58.4 +/- 6.9 pg/mL, P < 0.05), peaked at 12 POH, and still higher than that of NC group at 24 POH, while the serum level of TFPI showed no obvious difference in NC and CLP groups. Compared with that of NC group, TFPI levels obviously increased in CLP + CORM-2 group at 6, 12 POH (23.7 +/-3.5 ng/mL, 24.4 +/- 5.0 ng/mL vs 12.4 +/- 2.8 ng/mL, P < 0.05).</p><p><b>CONCLUSIONS</b>Exogenous CORM can obviously inhibit TF and NF-KB activity,decrease TF protein expression. Meanwhile, it can also decrease serum level of TF, and increase serum level of TFPI, preventing activation of procoagulant system, balancing procoagulant and anticoagulant system in sepsis.</p>