Interleukin-10 inhibits pulmonary NF-kappa B activation and lung injury induced by hepatic ischemia-reperfusion

Hiroyuki Yoshidome, Atsushi Kato, Michael J. Edwards, and Alex B. Lentsch

Department of Surgery, University of Louisville School of Medicine, Louisville, Kentucky 40292


    ABSTRACT
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Hepatic ischemia and reperfusion cause local and remote organ injury. This injury culminates from an integrated cascade of proinflammatory cytokines, chemokines, and adhesion molecules, many of which are regulated by the transcription factor nuclear factor-kappa B (NF-kappa B). The anti-inflammatory cytokine interleukin-10 (IL-10) has been shown to have inhibitory effects on NF-kappa B. The objective of the current study was to determine whether IL-10 could suppress pulmonary NF-kappa B activation and ensuing lung injury induced by hepatic ischemia-reperfusion. C57BL/6 mice underwent partial hepatic ischemia with or without intravenous administration of IL-10. Hepatic ischemia-reperfusion resulted in pulmonary NF-kappa B activation, increased mRNA expression of tumor necrosis factor-alpha (TNF-alpha ), and macrophage inflammatory protein-2 (MIP-2), as well as increased pulmonary neutrophil accumulation and lung edema. Administration of IL-10 suppressed lung NF-kappa B activation, reduced TNF-alpha and MIP-2 mRNA expression, and decreased pulmonary neutrophil recruitment and lung injury. The data suggest that IL-10 protects against hepatic ischemia and reperfusion-induced lung injury by inhibiting lung NF-kappa B activation and the resulting pulmonary production of proinflammatory mediators.

inflammation; tumor necrosis factor-alpha ; neutrophils; mice; nuclear factor-kappa B


    INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

HEPATIC ISCHEMIA-REPERFUSION injury is a complication of liver resectional surgery, liver transplantation, and hemorrhagic shock with fluid resuscitation and may lead to local and remote organ damage (17). There is a preponderance of literature suggesting that the pathogenesis of the resulting tissue injury in remote organs, such as the lung, is related to the hepatic generation of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha ). After hepatic ischemia and reperfusion, TNF-alpha initiates a mediator cascade in the lung, including upregulation of the C-X-C chemokines, epithelial neutrophil-activating protein ENA-78, macrophage inflammatory protein-2 (MIP-2), and KC, as well as increased pulmonary vascular expression of intercellular adhesion molecule-1 (ICAM-1) (3-5, 26). The coordinated effects of C-X-C chemokines and adhesion molecules result in pulmonary neutrophil accumulation and ensuing lung injury. The gene expression of these proinflammatory mediators, including TNF-alpha , C-X-C chemokines, and ICAM-1, is controlled at least in part by the transcription factor nuclear factor-kappa B (NF-kappa B) (2, 7, 24, 25). Although activation of NF-kappa B occurs in liver during ischemia-reperfusion injury (28), it is unknown whether NF-kappa B is activated in remote organs or whether remote organ NF-kappa B activation is important for the development of tissue injury.

Recent reports have demonstrated that the anti-inflammatory cytokine interleukin-10 (IL-10) has hepatoprotective effects during liver transplantation (27) and experimental liver injury induced by galactosamine and lipopolysaccharide (18, 23). IL-10 has also been shown to suppress experimental lung inflammatory injury induced by intrapulmonary deposition of IgG immune complexes through inhibitory effects on NF-kappa B activation (15). In the current studies, we sought to determine whether hepatic ischemia and reperfusion caused NF-kappa B activation in the lung. In addition, we evaluated the effects of IL-10 on the development of lung inflammatory injury induced by hepatic ischemia and reperfusion.


    MATERIALS AND METHODS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Hepatic ischemia and reperfusion injury model. Male C57BL/6 mice (Charles River Laboratories, Wilmington, MA) weighing 22-28 g were used in all experiments. This project was approved by the University of Louisville Animal Care and Use Committee and was in compliance with the guidelines of the National Institutes of Health. The model of partial hepatic ischemia and reperfusion employed was prepared as described previously (16). Briefly, mice were anesthetized with pentobarbital sodium (60 mg/kg ip). Mice received either sterile saline or recombinant murine IL-10 (1 µg; R&D Systems, Minneapolis, MN) via the lateral tail vein before induction of ischemia. A midline laparotomy was performed, and an atraumatic clip was used to interrupt the arterial and the portal venous blood supply to the cephalad lobes of the liver. After 90 min of partial hepatic ischemia, mice again received either sterile saline or IL-10 (1 µg) via the lateral tail vein and the clip was removed, initiating hepatic reperfusion. Sham control mice underwent the same protocol but without vascular occlusion. Mice were killed after the indicated periods of reperfusion, and lung tissues and blood samples were taken for analysis.

Myeloperoxidase assay. Lung myeloperoxidase (MPO) content was assessed by methods described previously (26). Briefly, lung tissue (50 mg) was homogenized in 2 ml of homogenization buffer (3.4 mM KH2HPO4 and 16 mM Na2HPO4, pH 7.4). After centrifugation for 20 min at 10,000 g, 10 volumes of resuspension buffer (43.2 mM KH2HPO4, 6.5 mM Na2HPO4, 10 mM EDTA, and 0.5% hexadecyltrimethylammonium, pH 6.0) were added to the pellet and the samples were sonicated for 10 s. After heating for 2 h at 60°C, the supernatant was reacted with 3,3',5,5'-tetramethylbenzidine (Sigma Chemical, St. Louis, MO) and read at 655 nm.

Lung edema. The extent of lung edema was measured by tissue wet-to-dry weight ratios. After dissection, lung samples were weighed and then placed in a drying oven at 55°C until a constant weight was obtained. In this determination, lung edema is represented by an increase in the wet-to-dry weight ratio.

Reverse transcription-polymerase chain reaction. Fresh lung samples were immediately frozen when the mice were killed. The extraction of total RNA was performed using a commercially available kit (Qiagen, Valencia, CA). A 1-µg aliquot of total lung RNA was reverse transcribed to cDNA using the Geneamp RNA polymerase chain reaction (PCR) protocol (Perkin-Elmer, Norwalk, CT) with random hexamers to prime the reverse transcription (RT) with an excess of deoxyribonucleotides. The cDNA products were amplified with 2.5 units of AmpliTaq DNA polymerase, 3 mM Mg2+, and 0.5 mM primer. After 2 min of initial melting at 95°C, the mixture was amplified for a total of 30 cycles with a three-step cycle process that began with melting at 95°C for 60 s, annealing at 59°C for 90 s, followed by extension at 72°C for 10 s. The final cycle was followed by 12 min of soaking at 72°C. Ten microliters of each RT-PCR were electrophoresed in a 3.5% agarose (GIBCO) gel and stained with ethidium bromide. RT-PCR amplification of a housekeeping gene (beta -actin) was performed to verify equal loading of RNA and cDNA in the RT-PCRs. Products of RT-PCRs were photographed.

PCR primers were as follows: MIP-2 sense, 5'-GAA CAA AGG CAA GGC TAA CTG A-3'; MIP-2 antisense, 5'-AAC ATA ACA ACA TCT GGG CAA T-3' (to give a 205-bp product, designed using OLIGO Software; NBI, Plymouth, MN; from GenBank accession no. 53798); TNF-alpha sense, 5'-AGC CCA CGT AGC AAA CCA CCA A-3'; TNF-alpha antisense, 5'-ACA CCC ATT CCC TTC ACA GAG CAA T-3' [to give a 446-bp product (15)]; beta -actin sense, 5'-GTG GGC CGC TCT AGG CAC CA-3'; beta -actin antisense, 5'-CGG TTG GCC TTA GGG TTC AGG GGG G-3' (to give a 245-bp product; Stratagene, La Jolla, CA).

NF-kappa B activation. Nuclear extracts of lung tissue were prepared by the method of Deryckere and Gannon (10). Protein concentrations were determined by bicinchoninic acid assay with trichloroacetic acid precipitation using BSA as a reference standard (Pierce, Rockford, IL). Double-stranded NF-kappa B consensus oligonucleotide (5'-AGTGAGGGGACTTTCCCAGGC-3'; Promega, Madison, WI) was end labeled with [gamma -32P]ATP (3,000 Ci/mmol at 10 mCi/ml; Amersham, Arlington Heights, IL). Binding reactions containing equal amounts of protein (20 µg) and 35 fmol (~50,000 counts/min, Cherenkov counting) of oligonucleotide were performed for 30 min in binding buffer [4% glycerol, 1 mM MgCl2, 0.5 mM EDTA, pH 8.0, 0.5 mM dithiothreitol, 50 mM NaCl, 10 mM Tris, pH 7.6, and 50 µg/ml poly(dI-dC); Pharmacia, Piscataway, NJ]. Reaction volumes were held constant to 15 µl. Reaction products were separated in a 4% polyacrylamide gel and analyzed by autoradiography.

Statistical analysis. All data are expressed as means ± SE. Data were analyzed with a one-way analysis of variance, and individual group means were then compared with a Student-Newman-Keuls test. Differences were considered significant when P < 0.05. For calculations of percent change, negative control values were subtracted from positive control and treatment group values.


    RESULTS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Activation of NF-kappa B in lung during hepatic ischemia-reperfusion injury. The time course of NF-kappa B activation in the lung during hepatic ischemia-reperfusion injury was determined by electrophoretic mobility shift assays of lung nuclear extracts (Fig. 1A). Autoradiograms were digitized and relative band intensity was determined using image-analysis software (Fig. 1B). In lungs from sham-operated controls, there was little NF-kappa B in nuclear extracts. Unexpectedly, nuclear translocation (activation) of NF-kappa B was observed after 90 min of hepatic ischemia but before reperfusion (0-h reperfusion). Lung NF-kappa B activation increased modestly over the 4-h time course studied, with maximal activation occurring 1 and 4 h after hepatic reperfusion.


View larger version (46K):
[in this window]
[in a new window]
 
Fig. 1.   Time course of pulmonary nuclear factor-kappa B (NF-kappa B) activation during hepatic ischemia-reperfusion. A: nuclear extracts from whole lung tissues were subjected to electrophoretic mobility shift assay (EMSA). B: image analysis of EMSA autoradiogram. Results are representative of 4 separate time-course experiments.

Effects of IL-10 on lung NF-kappa B activation and mRNA expression of TNF-alpha and MIP-2. Because lung NF-kappa B activation induced by hepatic ischemia and reperfusion was maximally activated within 1-4 h of hepatic reperfusion, we assessed the effects of exogenously administered IL-10 on lung NF-kappa B activation after 1 h of reperfusion (Fig. 2). As in previous experiments, lung NF-kappa B activation was increased by hepatic ischemia-reperfusion compared with sham controls. In mice receiving intravenous administration of IL-10, lung NF-kappa B activation was almost completely inhibited.


View larger version (67K):
[in this window]
[in a new window]
 
Fig. 2.   Effects of interleukin-10 (IL-10) on hepatic ischemia-reperfusion (I/R)-induced lung NF-kappa B activation. Nuclear translocation of NF-kappa B was assessed in nuclear extracts from lung tissue obtained from sham mice and mice undergoing hepatic ischemia and 1 h of reperfusion treated with saline (I/R) or IL-10 (I/R + IL-10).

To determine whether IL-10-mediated suppression of lung NF-kappa B activation was associated with decreased proinflammatory mediator generation, lung expression of mRNAs for TNF-alpha and MIP-2 was assessed in sham control mice and in mice undergoing hepatic ischemia and 1 or 4 h of reperfusion in the presence or absence of IL-10 treatment. Lung RNA extracts were analyzed by RT-PCR. Hepatic ischemia-reperfusion induced a small increase in TNF-alpha mRNA after 1 h of reperfusion (Fig. 3A). This increase was completely abrogated with the administration of IL-10 (Fig. 3A). Hepatic ischemia and 4 h of reperfusion induced a much larger increase in the expression of TNF-alpha mRNA, which was greatly reduced with administration of IL-10. Pulmonary expression of MIP-2 mRNA was modestly increased after 1 h of reperfusion (Fig. 3B). After 4 h of reperfusion, lung MIP-2 mRNA was greatly increased. Administration of IL-10 resulted in a slight decrease in MIP-2 mRNA expression after 1 h of reperfusion but more effectively inhibited MIP-2 mRNA expression after 4 h of reperfusion (Fig. 3B).


View larger version (24K):
[in this window]
[in a new window]
 
Fig. 3.   Effects of IL-10 on lung expression of tumor necrosis factor-alpha (TNF-alpha ; A) and macrophage inflammatory protein-2 (MIP-2; B) mRNAs. RT-PCR analysis of lung RNA extracts obtained from sham mice and mice undergoing hepatic ischemia-reperfusion treated with saline (I/R) or IL-10 (I/R + IL-10). MWM, Molecular-weight marker. Right panels show image analysis of RT-PCR results.

Effects of IL-10 on lung neutrophil recruitment and injury. We have previously shown in this model that pulmonary neutrophil accumulation and lung injury correlate with lung mRNA and serum levels of MIP-2 and TNF-alpha (26). The recruitment of neutrophils to lung and the development of lung injury occur after 3-4 h of hepatic reperfusion (26). Therefore, we assessed whether the reduction in TNF-alpha and MIP-2 mRNA expression by IL-10 was associated with decreased recruitment of neutrophils and lung injury after 4 h of hepatic reperfusion. Pulmonary neutrophil recruitment was determined by lung MPO content. Hepatic ischemia and 4 h of reperfusion caused significant increases in lung MPO content compared with the sham controls (Fig. 4). Intravenous administration of IL-10 reduced lung MPO content by 61% (P = 0.016). Lung edema, as measured by lung wet-to-dry weight ratio, was used as an index of lung injury. Hepatic ischemia and 4 h of reperfusion induced significant lung edema (Fig. 5). Administration of IL-10 suppressed increases in lung wet-to-dry weigh ratio by 72% (P = 0.004).


View larger version (18K):
[in this window]
[in a new window]
 
Fig. 4.   Effects of IL-10 on lung neutrophil recruitment induced by hepatic ischemia and reperfusion. Myeloperoxidase (MPO) content in lung tissues from sham control mice and mice undergoing hepatic ischemia and 4 h of reperfusion in the presence or absence of IL-10 was determined. For all groups n = 5.



View larger version (21K):
[in this window]
[in a new window]
 
Fig. 5.   Effects of IL-10 on lung edema induced by hepatic ischemia and reperfusion. Lung edema was determined by tissue wet-to-dry weight ratios in sham control mice and mice undergoing hepatic ischemia and 4 h of reperfusion in the presence or absence of IL-10. For all groups n = 5.


    DISCUSSION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Hepatic injury induced by ischemia and reperfusion is associated with two distinct phases. During the initial phase, activated Kupffer cells cause liver injury by releasing reactive oxygen species (12). After this initial oxidant-induced injury, Kupffer cells and hepatocytes produce proinflammatory cytokines, including TNF-alpha , which initiates a mediator cascade leading to hepatic neutrophil recruitment (6). The ensuing liver injury is mediated by oxidants and proteases released by sequestered neutrophils (11). It has been shown that activation of NF-kappa B occurs during the initial phase of hepatic injury (28). It is thought that NF-kappa B activation may be responsible at least in part for the increased hepatic production of proinflammatory cytokines during this phase. Remote organ injury caused by hepatic ischemia and reperfusion, such as that induced in the lung, is thought to be a result of liver-derived TNF-alpha . In fact, blockade of TNF-alpha using antibody neutralization greatly reduced hepatic ischemia and reperfusion-induced lung inflammatory injury in rats (6).

Unexpected were the findings that NF-kappa B was activated in the lung during the period of hepatic ischemia. The precise mechanism of this effect is unknown, but serum TNF-alpha is not increased at this time (Yoshidome and Lentsch, unpublished data). Our observations of increased lung NF-kappa B activation during hepatic ischemia are consistent with those of a recent report demonstrating increased lung production of C-X-C chemokines during hindlimb ischemia before reperfusion (1). In those studies, it was noted that there was systemic activation of complement during the ischemic period. Although complement activation products have been shown to augment pulmonary production of TNF-alpha and C-X-C chemokines (9, 20), these effects are not mediated by activation of NF-kappa B (9, 14). Alternatively, mediators could be released by the nonischemic liver in response to increased blood flow to that portion of the organ. However, there is no evidence of tissue injury in nonischemic lobes (16).

On the basis of our findings, increased NF-kappa B activation in lung after hepatic ischemia-reperfusion appears to lead to pulmonary expression of TNF-alpha and MIP-2. Activation of NF-kappa B in alveolar macrophages has been shown to be required for the intrapulmonary production of TNF-alpha and MIP-2, as well as development of lung injury induced by IgG immune complexes (13). IL-10 has been shown to suppress NF-kappa B in alveolar macrophages by preventing degradation of the NF-kappa B inhibitory protein Ikappa B-alpha (15). In the present studies, intravenous administration of IL-10 suppressed lung NF-kappa B activation induced by hepatic ischemia and reperfusion. These effects were associated with reduced lung expression of TNF-alpha and MIP-2 mRNAs, both of which are required for full development of lung injury after hepatic ischemia and reperfusion (3, 26). Reductions in TNF-alpha and MIP-2 corresponded to decreased lung neutrophil accumulation and reduced lung edema. Additionally, IL-10 may reduce neutrophil recruitment into the lung through effects on the pulmonary endothelium. IL-10 has been shown to prevent upregulation of lung vascular ICAM-1 during lung inflammation (19). Furthermore, recent studies showed that blockade of TNF-alpha decreases pulmonary vascular expression of the adhesion molecule ICAM-1, resulting in reduced lung recruitment of neutrophils (4). Like TNF-alpha and MIP-2, ICAM-1 is regulated at the transcriptional level by NF-kappa B (7). Although not investigated in the current study, it is likely that IL-10 reduces pulmonary vascular expression of ICAM-1 either directly through inhibition of NF-kappa B in endothelial cells or indirectly by suppression of TNF-alpha production as a contributing mechanism for reduction of lung neutrophil accumulation.

IL-10 has been shown to be beneficial in the setting of organ transplantation, including liver (8, 27) and heart (21, 22); in both cases, treatment with IL-10 increased allograft survival. The current studies demonstrate that IL-10 may also be protective in surgical or trauma-related organ injuries occurring secondary to hepatic ischemia-reperfusion. IL-10 reduced lung injury induced by hepatic ischemia and reperfusion. IL-10 decreased the extent of lung neutrophil recruitment in association with reductions in lung expression of TNF-alpha and MIP-2 mRNAs. These effects of IL-10 were accompanied by almost complete inhibition of NF-kappa B activation in the lung. Thus the current studies suggest that the use of agents such as IL-10 to target upstream components of the inflammatory response, such as NF-kappa B, may have therapeutic applications for organ injury caused by hepatic surgery, transplantation or hemorrhagic shock.


    ACKNOWLEDGEMENTS

This work was supported in part by grants from the Alliant Community Trust Fund and the Jewish Hospital Foundation.


    FOOTNOTES

The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

Address for reprint requests and other correspondence: A. B. Lentsch, Dept. of Surgery, Univ. of Louisville School of Medicine, J. G. Brown Cancer Center, Rm. 426, 529 South Jackson St., Louisville, KY 40292 (E-mail: ablent01{at}ulkyvm.louisville.edu).

Received 12 February 1999; accepted in final form 9 June 1999.


    REFERENCES
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

1.   Bless, N. M., R. L. Warner, V. A. Padgaonkar, A. B. Lentsch, B. J. Czermak, H. Schaml, H. P. Friedl, and P. A. Ward. Roles for C-X-C chemokines and C5a in lung injury after hindlimb ischemia-reperfusion. Am. J. Physiol. 276 (Lung Cell. Mol. Physiol. 20): L57-L63, 1999[Abstract/Free Full Text].

2.   Collart, M. A., P. Baeuerle, and P. Vassalli. Regulation of tumor necrosis factor alpha transcription in macrophages: involvement of four kappa B-like motifs and of constitutive and inducible forms of NF-kappa B. Mol. Cell. Biol. 10: 1498-1506, 1990[Medline].

3.   Colletti, L. M., G. D. Burtch, D. G. Remick, S. L. Kunkel, R. M. Strieter, K. S. Guice, K. T. Oldham, and D. A. Campbell, Jr. The production of tumor necrosis factor alpha and the development of a pulmonary capillary injury following hepatic ischemia/reperfusion. Transplantation 49: 268-272, 1990[Medline].

4.   Colletti, L. M., A. Cortis, N. Lukacs, S. L. Kunkel, M. Green, and R. M. Strieter. Tumor necrosis factor up-regulates intercellular adhesion molecule 1, which is important in the neutrophil-dependent lung and liver injury associated with hepatic ischemia and reperfusion in the rat. Shock 10: 182-191, 1998[Medline].

5.   Colletti, L. M., S. L. Kunkel, A. Walz, M. D. Burdick, R. G. Kunkel, C. A. Wilke, and R. M. Strieter. Chemokine expression during hepatic ischemia/reperfusion-induced lung injury in the rat. The role of epithelial neutrophil activating protein. J. Clin. Invest. 95: 134-141, 1995[Medline].

6.   Colletti, L. M., D. G. Remick, G. D. Burtch, S. L. Kunkel, R. M. Strieter, and D. A. Campbell, Jr. Role of tumor necrosis factor-alpha in the pathophysiologic alternations after hepatic ischemia/reperfusion injury in the rat. J. Clin. Invest. 85: 1936-1943, 1990[Medline].

7.   Collins, T., M. A. Read, A. S. Neish, M. Z. Whitley, D. Thanos, and T. Maniatis. Transcriptional regulation of endothelial cell adhesion molecules: NF-kappa B and cytokine-inducible enhancers. FASEB J. 9: 899-909, 1995[Abstract/Free Full Text].

8.   Conti, F., M. L. Boulland, K. Leroy-Viard, C. Chereau, B. Dousset, O. Soubrane, B. Weill, and Y. Calmus. Low level of interleukin 10 synthesis during liver allograft rejection. Lab. Invest. 78: 1281-1289, 1998[Medline].

9.   Czermak, B. J., A. B. Lentsch, N. M. Bless, H. Schmal, H. P. Friedl, and P. A. Ward. Synergistic enhancement of chemokine generation and lung injury by C5a or the membrane attack complex of complement. Am. J. Pathol. 154: 1513-1524, 1999[Abstract/Free Full Text].

10.   Deryckere, F., and F. Gannon. A one-hour minipreparation technique for extraction of DNA-binding proteins from animal tissues. Biotechniques 16: 405, 1994[Medline].

11.   Jaeschke, H., and C. W. Smith. Mechanisms of neutrophil-induced parenchymal cell injury. J. Leukoc. Biol. 61: 647-653, 1997[Abstract].

12.   Jaeschke, J., A. P. Bautista, Z. Spolarics, and J. J. Spitzer. Superoxide generation by Kupffer cells and priming of neutrophils during reperfusion after hepatic ischemia. Free Radic. Res. Commun. 15: 277-284, 1991[Medline].

13.   Lentsch, A. B., B. J. Czermak, N. M. Bless, N. V. Rooijen, and P. A. Ward. Essential role of alveolar macrophages in intrapulmonary activation of NF-kappa B. Am. J. Respir. Cell Mol. Biol. 20: 692-698, 1999[Abstract/Free Full Text].

14.   Lentsch, A. B., B. J. Czermak, N. M. Bless, and P. A. Ward. NF-kappa B activation during IgG immune complex-induced lung injury. Am. J. Pathol. 152: 1327-1336, 1998[Abstract].

15.   Lentsch, A. B., T. P. Shanley, V. Sarma, and P. A. Ward. In vivo suppression of NF-kappa B and preservation of Ikappa Balpha by interleukin-10 and interleukin-13. J. Clin. Invest. 100: 2443-2448, 1997[Abstract/Free Full Text].

16.   Lentsch, A. B., H. Yoshidome, W. G. Cheadle, F. N. Miller, and M. J. Edwards. Chemokine involvement in hepatic ischemia/reperfusion injury in mice: roles for macrophage inflammatory protein-2 and KC. Hepatology 27: 1172-1177, 1998[Medline].

17.   Liu, D. L., B. Jeppsson, C. H. Hakansson, and R. Odselius. Multiple-system organ damage resulting from prolonged hepatic inflow interruption. Arch. Surg. 131: 442-447, 1996[Abstract].

18.   Louis, H., O. Le Moine, M. O. Peny, B. Gulbis, F. Nisol, M. Goldman, and J. Deviere. Hepatoprotective role of interleukin 10 in galactosamine/lipopolysaccharide mouse liver injury. Gastroenterology 112: 935-942, 1997[Medline].

19.   Mulligan, M. S., M. L. Jones, A. A. Vaporciyan, M. C. Howard, and P. A. Ward. Protective effects of IL-4 and IL-10 against immune complex-induced lung injury. J. Immunol. 151: 5666-5674, 1993[Abstract/Free Full Text].

20.   Mulligan, M. S., E. Schmid, B. Beck-Schimmer, G. O. Till, H. P. Friedl, R. B. Brauer, T. E. Hugli, M. Miyasaka, R. L. Warner, K. J. Johnson, and P. A. Ward. Requirement and role of C5a in acute lung inflammatory injury in rats. J. Clin. Invest. 98: 503-512, 1996[Abstract/Free Full Text].

21.   Qin, L., K. D. Chavin, Y. Ding, H. Tahara, J. P. Favaro, J. E. Woodward, T. Suzuki, P. D. Robbins, M. T. Lotze, and J. S. Bromberg. Retrovirus-mediated transfer of viral IL-10 gene prolongs murine cardiac allograft survival. J. Immunol. 156: 2316-2323, 1996[Abstract].

22.   Raisanen-Sokolowski, A., T. Glysing-Jensen, and M. E. Russell. Leukocyte-suppressing influences of interleukin (IL)-10 in cardiac allografts: insights from IL-10 knockout mice. Am. J. Pathol. 153: 1491-1500, 1998[Abstract/Free Full Text].

23.   Santucci, L., S. Fiorucci, M. Chiorean, P. M. Brunori, F. M. Di Matteo, A. Sidoni, G. Migliorati, and A. Morelli. Interleukin 10 reduces lethality and hepatic injury induced by lipopolysaccharide in galactosamine-sensitized mice. Gastroenterology 111: 736-744, 1996[Medline].

24.   Stein, B., and A. S. J. Baldwin. Distinct mechanisms for regulation of the interleukin-8 gene involve synergism and cooperativity between C/EBP and NF-kappa B. Mol. Cell. Biol. 13: 7191-7198, 1993[Abstract].

25.   Widmer, U., K. R. Manogue, A. Cerami, and B. Sherry. Genomic cloning and promoter analysis of macrophage inflammatory protein (MIP)-2, MIP-1 alpha, and MIP-1 beta, members of the chemokine superfamily of proinflammatory cytokines. J. Immunol. 150: 4996-5012, 1993[Abstract/Free Full Text].

26.   Yoshidome, H., A. B. Lentsch, W. G. Cheadle, F. N. Miller, and M. J. Edwards. Enhanced pulmonary expression of CXC chemokines during hepatic ischemia/reperfusion-induced lung injury in mice. J. Surg. Res. 81: 33-37, 1999[Medline].

27.   Zou, X. M., A. Yagihashi, K. Hirata, T. Tsuruma, T. Matsuno, K. Tarumi, K. Asanuma, and N. Watanabe. Downregulation of cytokine-induced neutrophil chemoattractant and prolongation of rat liver allograft survival by interleukin-10. Surg. Today 28: 184-191, 1998[Medline].

28.   Zwacka, R. M., Y. Zhang, W. Zhou, J. Halldorson, and J. F. Engelhardt. Ischemia/reperfusion injury in the liver of BALB/c mice activates AP-1 and nuclear factor kappa B independently of Ikappa B degradation. Hepatology 28: 1022-1030, 1998[Medline].


Am J Physiol Lung Cell Mol Physiol 277(5):L919-L923
0002-9513/99 $5.00 Copyright © 1999 the American Physiological Society