CINC-1 Levels Research Articles

Activation of hepatic macrophage contributes to hepatic necrosis after post-ischemic reperfusion in alcoholic fatty liver

Fatty livers are vulnerable to ischemia/reperfusion (I/R) injury. We investigated the role of hepatic macrophages in the I/R injury in the fatty liver. Rats with alcoholic or nonalcoholic fatty liver were subjected to hepatic warm ischemia for 30 min. A bolus of gadolinium chloride (GdCl 3) was injected intravenously twice before I/R to block hepatic macrophage activity. Alcoholic fatty liver developed more extensive hepatic necrosis with neutrophil infiltration in association with a higher production of cytokine-induced neutrophil chemoattractant (CINC)-1, a potent neutrophil chemokine in rat, after I/R than the nonalcoholic fatty liver or control liver without steatosis. Hepatic apoptosis after I/R increased to a similar degree (3-fold) in each of the two fatty liver models, compared with the control liver. Alcoholic fatty liver exposed to I/R showed a rapid increase in nuclear factor-kappa B (NF-κB) binding activity. The GdCl 3 pretreatment significantly reduced NF-κB binding activity, CINC-1 level and necrosis in alcoholic fatty liver, despite no significant decrease in the extent of apoptosis. Our results suggest that the activation of hepatic macrophages in alcoholic fatty liver may contribute to hepatic necrosis after I/R, and that the apoptosis might be less dependent on the macrophage activity.

Gender dimorphism in neutrophil priming and activation following trauma-hemorrhagic shock

Although gender influences T-cell, macrophage and organ functions following trauma-hemorrhage and resuscitation (T-H), it remains unknown whether it also influences polymorphonuclear cell (PMN) activity under such conditions. To study this, proestrus female and male Sprague-Dawley rats underwent trauma-hemorrhage followed by fluid resuscitation. Circulating PMNs were assessed for superoxide (O2-) and elastase production and tissues were analyzed for myeloperoxidase (MPO) activity and TBARS (thiobarbituric acid reactive substances) as a marker of oxidative injury, at 2 and 24 h after resuscitation. PMA stimulated O2- production was not influenced by T-H or gender. In contrast, fMLP-stimulated O2- and LPS-stimulated elastase release by PMNs from male T-H rats was greater than that of females. A significant MPO activity and TBARS in tissues of both male and female rats was induced; however, MPO activity and TBARS levels were higher in males following T-H. Levels of the chemokine CINC-1 were elevated in the lungs of male, but not of proestrus females after T-H. Thus, decreased PMN priming and activation in proestrus females, compared to males, occurs following T-H resulting in decreased cellular injury and organ damage that is likely to contribute to improved outcome under those conditions.

Prophylactic effect of JTE-607 on LPS-induced acute lung injury in rats with CINC-1 inhibition.

JTE-607, a multiple cytokine inhibitor, was evaluated in lipopolysaccharide (LPS)-induced acute lung injury in rats in vivo and in vitro. LPS instillation into airways of rats was performed. JTE-607 at 3-30 mg/kg and dexamethasone at 3 mg/kg were administered intravenously at 10 min and 0 min for JTE-607, and 60 min for dexamethasone prior to the LPS instillation (n = 8). Cytokine-induced neutrophil chemoattractant (CINC)-1 level and myeloperoxidase (MPO) activity in lung were measured at 4 h after LPS instillation, and at 24 h for lung wet weight measurement and histological study. LPS-induced CINC-1 production by rat alveolar macrophages were also measured in vitro. JTE-607 and dexamethasone showed a significant reduction of increased CINC-1 level and MPO activity in lung after LPS treatment in vivo. Increased wet weight was also significantly inhibited. Histological studies revealed that JTE-607 and dexamethasone significantly inhibited LPS-induced accumulation of peribronchial neutrophils and eosinophils, and perivascular edema. JTE-607 and dexamethasone suppressed CfNC-1 synthesis by rat alveolar macrophages in vitro with IC50 values of 12.4 microM and 2.3 nM, respectively. These results indicate that JTE-607 has an inhibitory effect on LPS-induced rat lung inflammation in parallel with CINC-1 reduction. The effect of JTE-607 was suggested to be through direct inhibition of CINC-1 production from rat alveolar macrophages. JTE-607 may thus be efficacious in cytokine-mediated lung inflammation such as acute respiratory distress syndrome.

Effects of neutrophil elastase inhibitor (ONO-5046) on lung injury after intestinal ischemia-reperfusion

The underlying mechanisms of lung endothelial injury after intestinal ischemia-reperfusion (I/R) injury are not fully known. Here we investigated the effects of posttreatment with a neutrophil elastase inhibitor (NEI; ONO-5046) on lung injury after intestinal I/R injury in a rat model. Intestinal I/R was produced by 90 min of ischemia followed by either 60 or 240 min of reperfusion. For all experimental groups, the endothelial permeability index increased, neutrophil H(2)O(2) production increased in the pulmonary vasculature blood, neutrophil counts increased in bronchoalveolar lavage fluid (BALF), and the cytokine-induced neutrophil chemoattractant (CINC)-1 and CINC-3 levels were increased in BALF after 240 min (P < 0.01). In rats treated with NEI from 60 min after reperfusion, the lung endothelial permeability index was significantly reduced (P < 0.05), whereas neutrophil H(2)O(2) production in pulmonary vasculature blood and neutrophil count in BALF were significantly suppressed by NEI (P < 0.05 and P < 0.01, respectively). In addition, NEI significantly suppressed the increase of CINC-1 and CINC-3 levels in BALF (P < 0.05). Our study clearly indicates that posttreatment with NEI reduces neutrophil activation in the pulmonary vessels and neutrophil accumulation in the lungs and suggests that ONO-5046, even when administered after the primary intestinal insult, can prevent the progression of lung injury associated with intestinal I/R.

The role of cytokine‐induced neutrophil chemoattractant‐1 in neutrophil‐mediated remote lung injury after intestinal ischaemia/reperfusion in rats

Remote lung injury is induced by ischaemia/reperfusion (I/R) of the gastrointestinal tract and the liver following hypovolaemic shock. In the present study, the role of cytokine-induced neutrophil chemoattractant (CINC), a member of the interleukin (IL)-8 family, in neutrophil-mediated remote lung injury following intestinal I/R was investigated in anaesthetized rats. The I/R group was subjected to 60 min of occlusion of the superior mesenteric artery with laparotomy, followed by 240 min of intestinal reperfusion. The sham-operated (sham) group was subjected to the same procedures with the exception of intestinal I/R. In the I/R group, the permeability index of the lung, the neutrophil count in pulmonary vascular lavage fluid and bronchoalveolar lavage fluid (BALF), lung myeloperoxidase activity and neutrophil oxidative production were all significantly greater than those in the sham group. Cytokine-induced neutrophil chemoattractant-1 levels in blood and BALF were significantly increased at 240 min after intestinal reperfusion. There was a significant relationship between neutrophils in BALF and CINC-1 level in BALE CONCLUSION: These findings suggest that intestinal reperfusion was associated with activation and accumulation of neutrophils in the lung and resulted in remote lung injury with increased microvascular permeability. Thus, CINC-1 in BALF may induce neutrophil migration from the pulmonary vessels to the interstitium and alveolar spaces in remote lung injury after intestinal I/R.

Cytokine-induced neutrophil chemoattractant in a rat model of lipopolysaccharide-induced acute lung injury.

To elucidate the role of major chemotactic factors, cytokine-induced neutrophil chemoattractant (CINC), leukotriene B4 (LTB4) and C5a in lipopolysaccharide (LPS)-induced acute lung injury in rat, we employed three reagents: anti-CINC-1 antibody, an LTB4 receptor antagonist (ONO-4057) and an anti-complementary agent (K-76COONa). Rats were divided into five groups: (1)control group; (2) LPS group, which received intratracheal instillation of LPS (100 microg/kg); (3) Anti-CINC group, which received intratracheal coinstillation of LPS with anti-CINC-1 antibody (1 mg/kg); (4) LTB4-Ra group, which received intravenous ONO-4057 (10 mg/kg) prior to intratracheal LPS; (5) Anti-C5a group, which received intravenous K-76COONa (100 mg/kg) prior to intratracheal LPS. The number of neutrophils in bronchoalveolar lavage (BAL) fluids 6 h after LPS instillation was significantly reduced in the Anti-CINC group, however, no reduction was found in either the LTB4-Ra group or Anti-C5a group. The levels of CINC-1, CINC-2alpha and CINC-3 in BAL fluids were significantly higher in the LPS group than in the saline-instilled control group. In vitro, the production of CINC-1 and CINC-3 from LPS-stimulated macrophages was significantly elevated compared to unstimulated macrophages 6 h later. The increase in CINC-2alpha production was markedly less than that of CINC-1 or CINC-3. These results indicate that CINCs, especially CINC-1 and CINC-3 play an important role in the recruitment of neutrophils to the lung in LPS-induced acute lung injury.

炎症性サイトカインのｉｎ ｖｉｖｏにおける作用とその産生制御 ラット胸膜炎モデルを用いた検討

To assess roles of cytokines in the inflammatory process, we examined local production of proinflammatory cytokines such as tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-6 and cytokine-induced neutrophil chemoattractant-1 (CINC-1, a member of the CXC chemokine in rats), in the exudates of carrageenin induced pleurisy of rats. All four cytokine levels began to increase at 1-2h, and their production preceded the influx of neutrophils, and peaked at 3-5h, whereas the exudate volume and leukocyte number still continued to increase. When recombinant human (rh) TNFα or rhlL-1α was injected into the pleural cavity, the pleural CINC-1 level rapidly increased, preceding the neutrophil infiltration. A sharp rise in IL-6 level in the plasma, followed by an increase in T-kininogen, an acute-phase protein, was also observed. Intrapleural injection of rhIL-8 or CINC-1 caused neutrophil infiltration in a protein synthesis independent manner. Treatment with indomethacin, an anti-inflammatory drug, increased TNF and IL-1 levels but suppressed IL-6 level in the exudate of rats with pleurisy, whereas dexamethasone suppressed all these cytokine levels. Prostaglandins (PGs) and dibutyryl cAMP reduced the production of TNF and IL-1, while they increased that of IL-6 and CINC-1 from rat pleural resident cells. The above results clearly indicate that proinflammatory cytokines, TNF, IL-1, IL-6 and CINC-1 were produced locally. Moreover, these results suggest that these proinflammatory cytokine production could be regulated by inflammatory stimuli and also by other mediators such as PGE2 and PGI2 concomitantly produced in the inflammatory sites.