Limits...
Cytokines induce small intestine and liver injury after renal ischemia or nephrectomy.

Park SW, Chen SW, Kim M, Brown KM, Kolls JK, D'Agati VD, Lee HT - Lab. Invest. (2010)

Bottom Line: Small intestine histology after AKI showed profound villous lacteal capillary endothelial apoptosis, disruption of vascular permeability and epithelial necrosis.Small intestine appears to be the source of IL-17A, as IL-17A levels were higher in the portal circulation and small intestine compared with the levels measured from the systemic circulation and liver.Modulation of the inflammatory response and cytokine release in the small intestine after AKI may have important therapeutic implications in reducing complications arising from AKI.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, New York, NY 10032-3784, USA.

ABSTRACT
Patients with acute kidney injury (AKI) frequently suffer from extra-renal complications including hepatic dysfunction and systemic inflammation. We aimed to determine the mechanisms of AKI-induced hepatic dysfunction and systemic inflammation. Mice subjected to AKI (renal ischemia reperfusion (IR) or nephrectomy) rapidly developed acute hepatic dysfunction and suffered significantly worse hepatic IR injury. After AKI, rapid peri-portal hepatocyte necrosis, vacuolization, neutrophil infiltration and pro-inflammatory mRNA upregulation were observed suggesting an intestinal source of hepatic injury. Small intestine histology after AKI showed profound villous lacteal capillary endothelial apoptosis, disruption of vascular permeability and epithelial necrosis. After ischemic or non-ischemic AKI, plasma TNF-α, IL-17A and IL-6 increased significantly. Small intestine appears to be the source of IL-17A, as IL-17A levels were higher in the portal circulation and small intestine compared with the levels measured from the systemic circulation and liver. Wild-type mice treated with neutralizing antibodies against TNF-α, IL-17A or IL-6 or mice deficient in TNF-α, IL-17A, IL-17A receptor or IL-6 were protected against hepatic and small intestine injury because of ischemic or non-ischemic AKI. For the first time, we implicate the increased release of IL-17A from small intestine together with induction of TNF-α and IL-6 as a cause of small intestine and liver injury after ischemic or non-ischemic AKI. Modulation of the inflammatory response and cytokine release in the small intestine after AKI may have important therapeutic implications in reducing complications arising from AKI.

Show MeSH

Related in: MedlinePlus

Increased small intestine necrosis, apoptosis and inflammation after ischemic or non-ischemic AKIA. Representative photomicrographs of ileum from 5 experiments (hematoxylin and eosin staining, magnification 200X and 400X) of mice subjected to sham-operation (Sham), to 30 min. renal ischemia and 5 hrs of reperfusion (RIR) or to bilateral nephrectomy (BNx). Sham operated animals show normal-appearing intestine histology (left panel). As shown in middle upper panel, lower power images (200X) show full thickness of the ileal wall and villi that appear thickened, blunted and inflamed compared to sham. In addition, 5 hours after 30 min. renal IR or bilateral nephrectomy, severe intestine epithelial cell necrosis of villous lining cells and the development of a necrotic epithelial pannus (arrows) over the mucosal surface were observed (middle panel). Finally, we observed prominent capillary endothelial apoptosis within the central villi of ileum (right panel). Enlarged insert shows several apoptotic endothelial cells (red arrow heads) within a villus. B. Representative gel images and band intensity quantifications of semi-quantitative RT-PCR of the pro-inflammatory markers ICAM-1, TNF-α, IL-6, IL-17A, KC, MCP-1 and MIP-2 from ileum of mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Tissues were harvested 5 hrs after sham-operation or AKI induction. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM. C-E. Representative photomicrographs (400X, dark brown stain indicated by arrows) of 5 experiments of immunohistochemistry for neutrophils (C), macrophages (D) and T-lymphocytes (E) in the small intestine tissues harvested from mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR) 5 hrs prior. F. Representative fluorescence photomicrographs (of 5 experiments) of ileum sections illustrating apoptotic nuclei [terminal deoxynucleotidyl transferase biotin-dUTP nick end-labeling (TUNEL) fluorescence staining, 100X]. Mice were subjected to sham-operation, bilateral nephrectomy (BNx) or 30 min. renal ischemia reperfusion (RIR) 5 hrs prior. Enlarged insert (of ileum from mice subjected to 30 min. RIR) shows prominent capillary endothelial apoptosis. G. (Left) Representative photographs of small intestine tissues (duodenum, jejunum and ileum) isolated from mice subjected to sham-operation or to bilateral nephrectomy (BNx) 5 hrs prior and injected with Evans blue dye (EBD). (Right) Quantification of small intestine EBD extravasations in mice subjected to sham-operation (N=6), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Five hrs after surgery, EBD was extracted in formamide and the amount of extravasated EBD in the intestine was calculated against a standard curve. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2991383&req=5

Figure 6: Increased small intestine necrosis, apoptosis and inflammation after ischemic or non-ischemic AKIA. Representative photomicrographs of ileum from 5 experiments (hematoxylin and eosin staining, magnification 200X and 400X) of mice subjected to sham-operation (Sham), to 30 min. renal ischemia and 5 hrs of reperfusion (RIR) or to bilateral nephrectomy (BNx). Sham operated animals show normal-appearing intestine histology (left panel). As shown in middle upper panel, lower power images (200X) show full thickness of the ileal wall and villi that appear thickened, blunted and inflamed compared to sham. In addition, 5 hours after 30 min. renal IR or bilateral nephrectomy, severe intestine epithelial cell necrosis of villous lining cells and the development of a necrotic epithelial pannus (arrows) over the mucosal surface were observed (middle panel). Finally, we observed prominent capillary endothelial apoptosis within the central villi of ileum (right panel). Enlarged insert shows several apoptotic endothelial cells (red arrow heads) within a villus. B. Representative gel images and band intensity quantifications of semi-quantitative RT-PCR of the pro-inflammatory markers ICAM-1, TNF-α, IL-6, IL-17A, KC, MCP-1 and MIP-2 from ileum of mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Tissues were harvested 5 hrs after sham-operation or AKI induction. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM. C-E. Representative photomicrographs (400X, dark brown stain indicated by arrows) of 5 experiments of immunohistochemistry for neutrophils (C), macrophages (D) and T-lymphocytes (E) in the small intestine tissues harvested from mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR) 5 hrs prior. F. Representative fluorescence photomicrographs (of 5 experiments) of ileum sections illustrating apoptotic nuclei [terminal deoxynucleotidyl transferase biotin-dUTP nick end-labeling (TUNEL) fluorescence staining, 100X]. Mice were subjected to sham-operation, bilateral nephrectomy (BNx) or 30 min. renal ischemia reperfusion (RIR) 5 hrs prior. Enlarged insert (of ileum from mice subjected to 30 min. RIR) shows prominent capillary endothelial apoptosis. G. (Left) Representative photographs of small intestine tissues (duodenum, jejunum and ileum) isolated from mice subjected to sham-operation or to bilateral nephrectomy (BNx) 5 hrs prior and injected with Evans blue dye (EBD). (Right) Quantification of small intestine EBD extravasations in mice subjected to sham-operation (N=6), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Five hrs after surgery, EBD was extracted in formamide and the amount of extravasated EBD in the intestine was calculated against a standard curve. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM.

Mentions: Small intestine histology was also assessed 5 hrs after bilateral nephrectomy in H&E stained sections which demonstrated profound villous endothelial cell apoptosis (right panel, Figure 6A and magnified insert), individual epithelial cell necrosis of villous lining cells and the development of a necrotic epithelial pannus over the mucosal surface (middle panel of Figure 6A). We also observed congestion of villous capillaries and swelling and blunting of villi due to villous mononuclear cell mucosal inflammation and edema. Histological changes were qualitatively similar but inflammatory changes were more severe in the ileum than the jejunum (Supplemental Figure 3). Similar intestine histopathology was observed after 30 min. renal IR (Figure 6A, middle panel) or unilateral nephrectomy (Supplemental Figure 4). In addition, we demonstrate increased pro-inflammatory mRNA (ICAM-1, TNF-α, IL-6, IL-17A, KC, MCP-1 and MIP-2) expression in the small intestine after (30 min. renal IR) or non-ischemic (bilateral nephrectomy) AKI with RTPCR (Figure 6B). We also demonstrate significant influx of neutrophils (Figure 6C), macrophages (Figure 6D) and T-lymphocytes (Figure 6E) into the small intestine compared to the sham-operated mice 5 hrs after 30 min. renal IR or bilateral nephrectomy in mice. Neutrophil infiltration was scant in the intestines from sham-operated mice; however, infiltration was easily visible in the small intestines from mice subjected to ischemic (30 min. renal IR) or non-ischemic (bilateral nephrectomy) AKI.


Cytokines induce small intestine and liver injury after renal ischemia or nephrectomy.

Park SW, Chen SW, Kim M, Brown KM, Kolls JK, D'Agati VD, Lee HT - Lab. Invest. (2010)

Increased small intestine necrosis, apoptosis and inflammation after ischemic or non-ischemic AKIA. Representative photomicrographs of ileum from 5 experiments (hematoxylin and eosin staining, magnification 200X and 400X) of mice subjected to sham-operation (Sham), to 30 min. renal ischemia and 5 hrs of reperfusion (RIR) or to bilateral nephrectomy (BNx). Sham operated animals show normal-appearing intestine histology (left panel). As shown in middle upper panel, lower power images (200X) show full thickness of the ileal wall and villi that appear thickened, blunted and inflamed compared to sham. In addition, 5 hours after 30 min. renal IR or bilateral nephrectomy, severe intestine epithelial cell necrosis of villous lining cells and the development of a necrotic epithelial pannus (arrows) over the mucosal surface were observed (middle panel). Finally, we observed prominent capillary endothelial apoptosis within the central villi of ileum (right panel). Enlarged insert shows several apoptotic endothelial cells (red arrow heads) within a villus. B. Representative gel images and band intensity quantifications of semi-quantitative RT-PCR of the pro-inflammatory markers ICAM-1, TNF-α, IL-6, IL-17A, KC, MCP-1 and MIP-2 from ileum of mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Tissues were harvested 5 hrs after sham-operation or AKI induction. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM. C-E. Representative photomicrographs (400X, dark brown stain indicated by arrows) of 5 experiments of immunohistochemistry for neutrophils (C), macrophages (D) and T-lymphocytes (E) in the small intestine tissues harvested from mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR) 5 hrs prior. F. Representative fluorescence photomicrographs (of 5 experiments) of ileum sections illustrating apoptotic nuclei [terminal deoxynucleotidyl transferase biotin-dUTP nick end-labeling (TUNEL) fluorescence staining, 100X]. Mice were subjected to sham-operation, bilateral nephrectomy (BNx) or 30 min. renal ischemia reperfusion (RIR) 5 hrs prior. Enlarged insert (of ileum from mice subjected to 30 min. RIR) shows prominent capillary endothelial apoptosis. G. (Left) Representative photographs of small intestine tissues (duodenum, jejunum and ileum) isolated from mice subjected to sham-operation or to bilateral nephrectomy (BNx) 5 hrs prior and injected with Evans blue dye (EBD). (Right) Quantification of small intestine EBD extravasations in mice subjected to sham-operation (N=6), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Five hrs after surgery, EBD was extracted in formamide and the amount of extravasated EBD in the intestine was calculated against a standard curve. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2991383&req=5

Figure 6: Increased small intestine necrosis, apoptosis and inflammation after ischemic or non-ischemic AKIA. Representative photomicrographs of ileum from 5 experiments (hematoxylin and eosin staining, magnification 200X and 400X) of mice subjected to sham-operation (Sham), to 30 min. renal ischemia and 5 hrs of reperfusion (RIR) or to bilateral nephrectomy (BNx). Sham operated animals show normal-appearing intestine histology (left panel). As shown in middle upper panel, lower power images (200X) show full thickness of the ileal wall and villi that appear thickened, blunted and inflamed compared to sham. In addition, 5 hours after 30 min. renal IR or bilateral nephrectomy, severe intestine epithelial cell necrosis of villous lining cells and the development of a necrotic epithelial pannus (arrows) over the mucosal surface were observed (middle panel). Finally, we observed prominent capillary endothelial apoptosis within the central villi of ileum (right panel). Enlarged insert shows several apoptotic endothelial cells (red arrow heads) within a villus. B. Representative gel images and band intensity quantifications of semi-quantitative RT-PCR of the pro-inflammatory markers ICAM-1, TNF-α, IL-6, IL-17A, KC, MCP-1 and MIP-2 from ileum of mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Tissues were harvested 5 hrs after sham-operation or AKI induction. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM. C-E. Representative photomicrographs (400X, dark brown stain indicated by arrows) of 5 experiments of immunohistochemistry for neutrophils (C), macrophages (D) and T-lymphocytes (E) in the small intestine tissues harvested from mice subjected to sham-operation (Sham), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR) 5 hrs prior. F. Representative fluorescence photomicrographs (of 5 experiments) of ileum sections illustrating apoptotic nuclei [terminal deoxynucleotidyl transferase biotin-dUTP nick end-labeling (TUNEL) fluorescence staining, 100X]. Mice were subjected to sham-operation, bilateral nephrectomy (BNx) or 30 min. renal ischemia reperfusion (RIR) 5 hrs prior. Enlarged insert (of ileum from mice subjected to 30 min. RIR) shows prominent capillary endothelial apoptosis. G. (Left) Representative photographs of small intestine tissues (duodenum, jejunum and ileum) isolated from mice subjected to sham-operation or to bilateral nephrectomy (BNx) 5 hrs prior and injected with Evans blue dye (EBD). (Right) Quantification of small intestine EBD extravasations in mice subjected to sham-operation (N=6), bilateral nephrectomy (BNx) or 30 min. renal IR (RIR). Five hrs after surgery, EBD was extracted in formamide and the amount of extravasated EBD in the intestine was calculated against a standard curve. *P<0.05 vs. sham-operated mice. Error bars represent 1 SEM.
Mentions: Small intestine histology was also assessed 5 hrs after bilateral nephrectomy in H&E stained sections which demonstrated profound villous endothelial cell apoptosis (right panel, Figure 6A and magnified insert), individual epithelial cell necrosis of villous lining cells and the development of a necrotic epithelial pannus over the mucosal surface (middle panel of Figure 6A). We also observed congestion of villous capillaries and swelling and blunting of villi due to villous mononuclear cell mucosal inflammation and edema. Histological changes were qualitatively similar but inflammatory changes were more severe in the ileum than the jejunum (Supplemental Figure 3). Similar intestine histopathology was observed after 30 min. renal IR (Figure 6A, middle panel) or unilateral nephrectomy (Supplemental Figure 4). In addition, we demonstrate increased pro-inflammatory mRNA (ICAM-1, TNF-α, IL-6, IL-17A, KC, MCP-1 and MIP-2) expression in the small intestine after (30 min. renal IR) or non-ischemic (bilateral nephrectomy) AKI with RTPCR (Figure 6B). We also demonstrate significant influx of neutrophils (Figure 6C), macrophages (Figure 6D) and T-lymphocytes (Figure 6E) into the small intestine compared to the sham-operated mice 5 hrs after 30 min. renal IR or bilateral nephrectomy in mice. Neutrophil infiltration was scant in the intestines from sham-operated mice; however, infiltration was easily visible in the small intestines from mice subjected to ischemic (30 min. renal IR) or non-ischemic (bilateral nephrectomy) AKI.

Bottom Line: Small intestine histology after AKI showed profound villous lacteal capillary endothelial apoptosis, disruption of vascular permeability and epithelial necrosis.Small intestine appears to be the source of IL-17A, as IL-17A levels were higher in the portal circulation and small intestine compared with the levels measured from the systemic circulation and liver.Modulation of the inflammatory response and cytokine release in the small intestine after AKI may have important therapeutic implications in reducing complications arising from AKI.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, New York, NY 10032-3784, USA.

ABSTRACT
Patients with acute kidney injury (AKI) frequently suffer from extra-renal complications including hepatic dysfunction and systemic inflammation. We aimed to determine the mechanisms of AKI-induced hepatic dysfunction and systemic inflammation. Mice subjected to AKI (renal ischemia reperfusion (IR) or nephrectomy) rapidly developed acute hepatic dysfunction and suffered significantly worse hepatic IR injury. After AKI, rapid peri-portal hepatocyte necrosis, vacuolization, neutrophil infiltration and pro-inflammatory mRNA upregulation were observed suggesting an intestinal source of hepatic injury. Small intestine histology after AKI showed profound villous lacteal capillary endothelial apoptosis, disruption of vascular permeability and epithelial necrosis. After ischemic or non-ischemic AKI, plasma TNF-α, IL-17A and IL-6 increased significantly. Small intestine appears to be the source of IL-17A, as IL-17A levels were higher in the portal circulation and small intestine compared with the levels measured from the systemic circulation and liver. Wild-type mice treated with neutralizing antibodies against TNF-α, IL-17A or IL-6 or mice deficient in TNF-α, IL-17A, IL-17A receptor or IL-6 were protected against hepatic and small intestine injury because of ischemic or non-ischemic AKI. For the first time, we implicate the increased release of IL-17A from small intestine together with induction of TNF-α and IL-6 as a cause of small intestine and liver injury after ischemic or non-ischemic AKI. Modulation of the inflammatory response and cytokine release in the small intestine after AKI may have important therapeutic implications in reducing complications arising from AKI.

Show MeSH
Related in: MedlinePlus