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J Harkema

Publications and source records attributed to J Harkema.

8 recordsLinked to original sources
PubMed2001

Inhibition of Kupffer cells reduced CXC chemokine production and liver injury.

BACKGROUND: Cytokine production is a critical component of ischemia/reperfusion (IR) injury. In the liver, Kupffer cells produce cytokines and chemokines (i.e., cytokines with chemoattractant properties) that are important mediators in neutrophil recruitment and subsequent hepatocellular injury. Therefore, the role of Kupffer cells in chemokine production in hepatic IR injury was investigated. METHODS: Adult male C57BL/6 mice underwent 90 min of partial hepatic ischemia followed by various reperfusion times (i.e., 0, 1.5, 3, and 6 h). Gadolinium chloride (GC), which inhibits Kupffer cell activity, was administered to mice 48 and 24 h prior to ischemia. The control group received a corresponding volume of normal saline. Plasma levels of the cytokine macrophage inflammatory protein-2 (MIP-2), KC, and tumor necrosis factor (TNF)-alpha and liver mRNA were measured. Liver injury was assessed by plasma level of alanine transaminase (ALT) and histopathology. RESULTS: A reperfusion time-dependent liver injury occurred as indicated by increased levels of plasma ALT and histopathology. The injury was associated with increased plasma TNF-alpha, MIP-2, and KC and their hepatic mRNA expression and neutrophil infiltration into ischemic lobes of the liver. GC treatment significantly reduced the number of Kupffer cells as determined by the immunostained liver tissue sections. The extent of liver injury significantly decreased in GC-treated mice that were associated with decreased levels of plasma ALT, TNF-alpha, MIP-2, and KC and neutrophil infiltration. CONCLUSIONS: This study suggests that Kupffer cells are major contributors to cytokine production in hepatic IR and their modulation may serve as a potential target for therapeutic intervention.

Alanine Transaminase↗
PubMed2001

Hepatic ischemia/reperfusion injury in P-selectin and intercellular adhesion molecule-1 double-mutant mice.

Neutrophil adhesion and recruitment represents one of the early cellular events that occur during hepatic ischemia/reperfusion (IR) injury and plays a critical role in determining the extent of tissue damage. The adhesion molecules, such as selectins and intercellular adhesion molecules (ICAM), are important in mediating neutrophil-endothelial cell interactions and neutrophil emigration. The goal of this study was to evaluate the role of P-selectin and ICAM-1 in hepatic IR injury. Male wild-type and P-selectin/ICAM-1-deficient (P/I null) mice underwent 90 minutes of partial hepatic ischemia followed by reperfusion at various time points (0, 1.5, 3, and 6 hours). Reperfusion caused a time-dependent hepatocellular injury in both wild-type and P/I null mice as judged by plasma alanine aminotransferase (ALT) levels and liver histopathology examination. Although ALT levels were slightly lower in the P/I null mice compared with the wild-type mice the differences were not statistically significant. Neutrophil infiltration to the ischemic liver was observed in both mouse groups after 6 hours of reperfusion; however, the infiltration to the midzonal region of the ischemic liver was more pronounced in the wild-type group. This study suggests that hepatocellular injury induced after hepatic IR was independent of P-selectin and ICAM-1 in this model of acute inflammatory tissue injury.

Alanine Transaminase↗
PubMed1997

Linkage analysis of susceptibility to ozone-induced lung inflammation in inbred mice.

Exposures to the common air pollutant ozone (O3) cause decrements in pulmonary function and induce airway inflammation that is characterized by infiltration of polymorphonuclear neutrophils (PMNs; refs 1-4). Because of the impact that O3 may have on public health, it is critical to identify susceptibility factors. Highly reproducible, significant inter-individual variations in human pulmonary function responses to O3 support the hypothesis that genetic background is an important determinant. Initial analysis of PMN responses to O3 exposure in segregant populations derived from inflammation-prone (susceptible) C57BL/6J (B6) and inflammation-resistant C3H/HeJ (C3) inbred mice indicated that susceptibility was controlled by a locus we termed Inf2 (ref. 7). Subsequent analyses with recombinant inbred strains suggested that a more complex interaction of genes is involved. In this report, we identify a quantitative trait locus (QTL) for O3 susceptibility on chromosome 17. Candidate genes for the locus include Tnf, the gene encoding the pro-inflammatory cytokine tumour necrosis factor-alpha (Tnf). Antibody neutralization of the protein product of this putative candidate gene significantly protected against O3 injury in susceptible mice. These results strongly support linkage of O3 susceptibility to a QTL on chromosome 17 and Tnf as a candidate gene.

Animals↗
PubMed1995

Concurrent increases in the storage and release of mucin-like molecules by rat airway epithelial cells in response to bacterial endotoxin.

Mucus hypersecretion is a prominent response of the airways to bacterial infections. Recent findings showed that bacterial endotoxin, a lipopolysaccharide complex released from the bacterial cell wall, was able to induce at least one component of the hypersecretory response, i.e., an increase in the amount of stored epithelial mucosubstances (1, 2). The goal of the present study was to determine whether endotoxin also was capable of increasing mucosubstance release from cells. Based on evidence that human mucin antibodies A10G5 and B6E8 cross-reacted with rat mucin-like molecules, we used the antibodies in enzyme-linked immunosorbent assays (ELISA) to compare mucin concentrations in bronchoalveolar lavage (BAL) fluid from endotoxin-treated and control rats. Results showed that endotoxin treatment increased the amount of released mucin over that in controls 1.5-fold at 96 h and 2.5-fold at 168 h after instillation. Thus, these studies have defined the previously detected mucosubstances as mucin-like molecules and showed that endotoxin increases their release from, as well as their storage in, rat airway epithelium. Concurrent increases in storage and release suggest that endotoxin also stimulates mucin synthesis and/or stability.

Animals↗
PubMed1994

Consequences of prolonged inhalation of ozone on Fischer-344/N rats: collaborative studies. Part I: Content and cross-linking of lung collagen.

Male and female Fischer-344 rats were exposed either to filtered air (controls) or to 0.12, 0.5, or 1.0 parts per million (ppm)* ozone for six hours per day, five days per week, for 20 months. We examined collagen deposition in lung tissue from these animals to determine whether or not chronic exposure of rats to ozone causes pulmonary fibrosis, as defined biochemically. Several techniques were used to study collagen deposition in the lungs of the animals. These methods included biochemical quantification by analysis of 4-hydroxyproline in lung tissue hydrolysates. The hydroxylysine-derived cross-links in mature collagen were quantified to estimate biochemically the excess of fibrotic collagen in the lung tissue. Biochemical analysis indicated excess collagen in the female rats exposed to 0.5 or 1.0 ppm ozone. Collagen in the lungs of the females also contained relatively more hydroxylysine-derived cross-links than did the lung collagen from age-matched control animals that had breathed only filtered air. Exposure of Fischer-344 rats for 20 months to 0.5 or 1.0 ppm ozone was associated with excess fibrotic lung collagen deposition as defined histologically. In female rats, exposure was also associated with excess deposition as determined biochemically. There was no indication of any significant changes in the lungs of any of the rats exposed to 0.12 ppm ozone, but the number of animals in this group was far too small to conclude whether this was a true no-observable-effect level. We conclude that chronic exposure of rats for 20 months to ozone at concentrations of 0.5 ppm or above for six hours per day, five days per week, causes mild to moderate lung fibrosis, as defined histologically and, in female rats, biochemically. The significance of these observations with regard to health risks to humans chronically inhaling ozone at ambient levels in polluted air remains to be determined.

Air Pollutants↗
PubMed1993

Effects of 20 months of ozone exposure on lung collagen in Fischer 344 rats.

Fischer 344 rats were exposed to filtered air (controls) or to 0.12, 0.5, or 1.0 ppm of ozone for 6 h/day, 5 days/week, for 20 months. We examined lung collagen deposition and metabolism in tissue from these animals to determine whether chronic exposure of rats to ozone causes pulmonary fibrosis. We observed excess stainable collagen in the centriacinar region of lungs from the rats exposed to 0.5 or 1.0 ppm of ozone. Biochemical analysis indicated a slight, yet significant, excess collagen deposition in the female rats exposed to 0.5 or 1.0 ppm of ozone. Collagen in the lungs of the females also contained relatively more hydroxylysine-derived crosslinks than did lung collagen from age-matched control animals. No excess of type I procollagen mRNA could be appreciated by in situ hybridization in lungs of the rats exposed to 1.0 ppm of ozone for 20 months, although this mRNA was detected in occasional alveolar interstitial cells at 2 months of exposure to ozone under the same protocol. These findings indicate that chronic exposure of rats to ozone causes mild, persistent fibrosis. The significance of these observations with regard to human health risks of chronically inhaling ozone at ambient levels in polluted air remains to be determined.

Animals↗
PubMedDate not supplied

L-selectin blockade and liver function in rats after uncontrolled hemorrhagic shock.

Hemorrhagic shock (HS) and resuscitation can be seen as a global body ischemia-reperfusion (I/R) injury characterized by neutrophil infiltration and organ damage. Liver dysfunction occurs early after HS. Adhesion molecules are needed for the first steps ofneutrophil migration. Thus, the purpose of this study was to investigate the role of L-selectin in the liver after uncontrolled HS and resuscitation. Forty-eight Sprague Dawley rats were subjected to uncontrolled HS and resuscitation. Animals were divided into three groups: sham, uncontrolled HS and resuscitation, and uncontrolled HS and resuscitation with anti-L-selectin treatment. At 6 we evaluated liver injury tests, liver tissue myeloperoxidase (MPO), and liver histology. Survival was followed for 3 days and compared between groups. Statistical analysis included Fisher's exact test and one-way analysis of variance. Survival significantly increased from 30% in the control group to 60% in the treated group (p < .05). Hepatocellular and structural injury as well as neutrophil infiltration was significantly decreased in treated animals (p < .05). Thus, blockade of L-selectin resulted in decreased hepatocellular injury and increased survival in our model of uncontrolled HS. Selectins may be important therapeutic targets for blockade in the treatment of HS.

Animals↗