Excretion mass balance evaluation, metabolite profile analysis and metabolite identification in plasma and excreta after oral administration of [14C]-meloxicam to the male cat: preliminary study.

The objective of this study was to investigate the metabolic pathways and routes of excretion of oral meloxicam in the cat. [(14)C]-meloxicam was administered orally to three fasted male cats. Urine, faeces, vomit and cage washes were collected over the following 144 h period. Blood was collected predosing and at 3 and 12 h postdosing. Metabolites were identified by HPLC/MS/MS. When possible a metabolic structure was proposed for each metabolite detected. Only unchanged meloxicam was identified in plasma. Five major metabolites were detected in urine and four in faeces, which were identified by HPLC/MS/MS as products of oxidative metabolism. No conjugated metabolites were detected. Elimination occurred early (61% during the first 48 h). A total of 21% of the recovered dose was eliminated in urine (2% as unchanged meloxicam, 19% as metabolites) and 79% in the faeces (49% as unchanged meloxicam, 30% as metabolites). The results indicate that after oral administration the major route of excretion of meloxicam in the cat is faecal and that the main pathway of biotransformation of meloxicam in the cat is oxidation.

Negative regulation of Apo A-I gene expression by retinoic acid in rat hepatocytes maintained in a coculture system.

Rat hepatocytes cocultured with rat liver epithelial cells (RLEC) were used to investigate the influence of all-trans retinoic acid (RA) on the regulation of apolipoproteins (Apo) A-I and A-II gene expression, the major protein constituent of high-density lipoproteins. In contrast to rat hepatocytes in conventional primary culture, Apo A-I and Apo A-II gene expression remained high and stable for several days in parenchymal cells in coculture. Treatment of cocultured rat hepatocytes with RA resulted in a specific decrease in Apo A-I mRNA levels whereas no marked difference in Apo A-II mRNA levels was observed. Such a negative effect of RA was already detected as early as 2 days of treatment and was effective for the entire experimental period (6 days). As controls, RARbeta mRNA levels increased whereas those of GAPDH mRNA were not affected by the RA treatment. The decrease in Apo A-I mRNA levels was associated with lower amounts of Apo A-I secreted in the culture medium within day 1 of treatment. This effect required active transcription and protein synthesis. These results show that, contrary to primary pure hepatocyte cultures and hepatoma cell lines, cocultures of rat hepatocytes reproduce the in vivo results suggesting that only well differentiated hepatocytes may correctly respond to RA. Furthermore, they demonstrate that RA can directly act on hepatocytes and differently affect Apo A-I and Apo A-II gene expression.

Suppression of humoral immunization against encapsulated xenogeneic hepatocytes and prolongation of their function by 2-week cyclosporine treatment in the rat.

BACKGROUND: Xenogeneic liver transplantation may induce immune reactions not only against the grafted liver but also against the proteins that it synthesizes. We investigated whether 2-week cyclosporine treatment could suppress immunization and improve graft function in a xenogeneic hepatocyte transplantation model. METHODS: Free or encapsulated human hepatoma cells (HepG2) were cocultured for 28 days with splenocytes from Lewis rats or implanted for 60 days into the peritoneum of Lewis rats. RESULTS: Anti-HepG2 and antialbumin antibodies were detected in the supernatants of rat splenocytes that were cocultured with HepG2 cells and in the serum of rats that had undergone transplantation with HepG2 cells. Cyclosporine suppressed this antibody production both in vitro and in vivo. Human alpha-GST blood levels, which reflect hepatocyte injury, were low in cyclosporine-treated animals but high when encapsulated HepG2 cells were transplanted without cyclosporine therapy. Western blots revealed human albumin from day 3 to day 60 in the serum of rats treated with cyclosporine, but not after day 30 in untreated rats. CONCLUSIONS: Xenogeneic hepatocytes induce a humoral response that impairs their viability and function. A 2-week course of cyclosporine suppresses this immune response and improves graft function for up to 60 days.

Expression of the membrane attack complex of complement and its inhibitors during human liver allograft transplantation.

BACKGROUND/AIMS: In order to test the possible role of activated complement in human liver allograft rejection, we evaluated the expression of the membrane attack complex of complement, its soluble inhibitors clusterin and vitronectin and its membrane inhibitor protectin during the evolution of liver transplants. METHODS: An indirect immunoperoxidase technique was applied to biopsy specimens obtained from liver allografts in 16 patients without complications, nine with acute rejection, four with chronic rejection and five with biliary complications. RESULTS: Two types of membrane attack complex deposition were observed: (a) extracellular deposits in portal tracts and perisinusoidal matrix, associated with clusterin and vitronectin, similar to those found in the normal liver; and (b) intra-portal vascular deposits, devoid of clusterin and vitronectin. Vascular membrane attack complex deposition was detected in four clinically stable patients, three patients with chronic rejection and two patients with biliary complications. In clinically stable patients, vascular membrane attack complex deposition was restricted to large portal vessels and was detected in a minority of portal tracts. In patients with chronic rejection or biliary complications, vascular membrane attack complex deposition was detected along both large and small portal vessels and was present in the majority of portal tracts. Protectin induction on hepatocytes was detected in 33 cases. CONCLUSIONS: Our results suggest that membrane attack complex deposition is unlikely to play a major role in the pathogenesis of acute liver allograft rejection but may contribute to the vascular and biliary lesions observed in chronic rejection.