Porcine hepatocyte-Kupffer cell co-culture as an in vitro model for testing the efficacy of anti-inflammatory substances.

As Kupffer cells are highly involved in the regulation of hepatic inflammatory response, the main goal of this study was to improve and to characterize a hepatocyte-Kupffer cell co-culture of pig origin for modelling endotoxin-induced hepatic inflammation and for testing the efficacy of potential anti-inflammatory substances. This monolayer co-culture was prepared from primary isolated swine hepatocytes and Kupffer cells in the ratio of 6:1 and 2:1, mimicking different states of liver inflammation. The prepared cell cultures were characterized by immunohistochemical CD-68 detection. Lipopolysaccharide (LPS) challenge of both co-cultures resulted in elevated interleukin-8 (IL-8) and that of 6:1 co-cultures in increased IL-6 production with a higher extent than on hepatocyte monocultures, justifying the key role of Kupffer cells in pro-inflammatory cytokine production. LPS-induced IL-8 production was successfully attenuated by concomitant application of both sodium butyrate and terpinen-4-ol on hepatocyte monocultures, but not on co-cultures, demonstrating the importance of the presence of Kupffer cells in cell cultures as inflammatory models. Based on these initial data, the applied porcine primary hepatocyte-Kupffer cell co-culture is suggested to be a proper tool for in vitro investigations on liver physiology and hepatic inflammation in pigs and can be used as a useful model mimicking in vivo conditions in veterinary research.

Effects of dietary sodium butyrate on hepatic biotransformation and pharmacokinetics of erythromycin in chickens.

Butyrate, a commonly applied feed additive in poultry nutrition, can modify the expression of certain genes, including those encoding cytochrome P450 (CYP) enzymes. In comparative in vitro and in vivo experiments, the effect of butyrate on hepatic CYP genes was examined in primary cultures of chicken hepatocytes and in liver samples of chickens collected from animals that had been given butyrate as a feed additive. Moreover, the effect of butyrate on the biotransformation of erythromycin, a marker substance for the activity of enzymes of the CYP3A family, was investigated in vitro and in vivo. Butyrate increased the expression of the avian-specific CYP2H1 both in vitro and in vivo. In contrast, the avian CYP3A37 expression was decreased in hepatocytes following butyrate exposure, but not in the in vivo model. CYP1A was suppressed by butyrate in the in vitro experiments, and overexpressed in vivo in butyrate-fed animals. The concomitant incubation of hepatocytes with butyrate and erythromycin led to an increased CYP2H1 expression and a less pronounced inhibition of CYP3A37. In in vivo pharmacokinetic experiments, butyrate-fed animals given a single i.m. injection of erythromycin, a slower absorption phase (longer T(half-abs) and delayed T(max)) but a rapid elimination phase of this marker substrate was observed. Although these measurable differences were detected in the pharmacokinetics of erythromycin, it is unlikely that a concomitant application of sodium butyrate with erythromycin or other CYP substrates will cause clinically significant feed-drug interaction in chickens.

Infusion of various short chain fatty acids causes different changes in the blood glucose and insulin concentrations in growing lambs deprived of food overnight.

Glucose and insulin concentrations of jugular blood plasma were monitored in growing lambs over 8 h, following a 2 h infusion of acetate, propionate, n-butyrate, n-valerate and physiological saline into the ruminal vein. Propionate and especially n-valerate infusion significantly increased blood glucose concentration. n-Butyrate induced only a small increase of shorter duration, while acetate failed to exert a pronounced effect on the blood glucose level. SCFA infusion, except for acetate, raised the insulin level in the blood. Relative rise was closely correlated with the length of carbon chain of the SCFA, that is, n-valerate caused the largest elevation of the insulin level, followed by n-butyrate and propionate. At the same time, acetate failed to cause a marked influence on the insulin level. These results of insulin showed agreement with glucose concentration changes, with the exception of n-butyrate treatment, where the increase of plasma insulin concentrations after the infusion proved to be much larger than that of glucose, relative to the preinfusion value.

Effect of intraruminal butyrate infusion on the plasma insulin level in sheep.

After intraruminal infusion of butyrate to sheep at dose rates of 0.25, 0.5, 1 and 2 g sodium n-butyrate per kg body mass, butyrate concentration of the rumen fluid and total secreted insulin rose in direct proportion to the butyrate dose infused. The half-life of butyrate in the rumen was always longer than that of insulin. At 90 min after the infusion of 1 g butyrate per kg body mass, butyrate concentration in the ruminal papillae reached the level corresponding to an extracellular concentration that reduced cell division by 50% in vitro. It can be concluded that butyrate may be present in the ruminal papillae in concentrations inhibiting cell proliferation, simultaneously with the presence of blood plasma insulin concentrations stimulating the proliferation of ruminal epithelial cells.