Inhibition of effects of endogenously synthesized histamine disturbs in vitro human dendritic cell differentiation.

Histamine, a principal mediator in various physiological and pathological cell functions is synthesized from L-histidine exclusively by histidine decarboxylase, an enzyme, which is expressed in many tissues of mammalian organism. Histamine plays a role in various cellular functions, including cell differentiation. The aim of this study was to determine the presence and to characterize the role of the endogenously produced histamine during in vitro dendritic cell (DC) differentiation induced by interleukin-4 (IL-4) and granulocyte-monocyte colony stimulating factor (GM-CSF). The changes in intracellular histamine content, biosynthesis and gene expression of histidine decarboxylase were investigated during this process. One also studied how histamine receptor antagonists and a histamine synthesis blocker influence the expression of differentiation antigens on the DC during in vitro maturation. During in vitro differentiation parallel culture incubations were performed by adding H1 receptor antagonist triprolidine, H2 receptor antagonist tiotidine, the tamoxifene derivate DPPE which blocks the intracellular binding of histamine, and an irreversible blocker of histidine decarboxylase, alpha-fluoromethyl histamine (alpha-FMH). The results show simultaneous increase in both histidine decarboxylase level and histamine content during differentiation of elutriated monocytes toward DC. Both blockade of de novo histamine production (by alpha-FMH) and inhibition of histamine binding (by H1 and H2 receptor antagonists, triprolidine and tiotidine, respectively) markedly decreased CD40 expression and that of CD45 from the 3rd day of treatment. DPPE by disturbing intracellular interaction of histamine with cytochrome P-450 moieties was able to decrease the expression of CD45, CD86, HLA-DR, CD33, CD40 and CD11c. Based on the data it is suggested that endogenous histamine is actively synthesized during cytokine-induced in vitro DC differentiation. The functional relevance and autocrine and paracrine action of endogenously produced histamine is supported by the data showing that inhibition of histamine synthesis by HDC, blocking of histamine binding by both 'extracellular' histamine receptors (by specific antagonists, triprolidine and tiotidine) and 'intracellular' antagonists (DPPE) disturb the differentiation of DC. This conclusion is supported by the fact, that by the inhibition of histamine acting in an autocrine/paracrine way, the expression pattern of differentiation markers on DC is markedly changed.

Application of LC-MS analysis to the characterisation of the in vitro and in vivo metabolite profiles of RGH-1756 in the rat.

RGH-1756, 1-(2-methoxy-phenyl)-4-(4-[4-(6-imidazol[2,1-b] thiazolyl)-phenoxy]-butyl-4-(14)C)-piperazine dimethane is a novel atypical antipsychotic drug candidate of Gedeon Richter Ltd. The metabolic pathways of the compound have been investigated by profiling the metabolites present in plasma, bile, and faeces samples of rats treated with (14)C-RGH-1756. The metabolites formed in vitro by rat liver microsomes have also been analysed. Good separation of the compounds has been achieved by gradient HPLC method on Zorbax/Bonus RP-C18 column. Radiometry and mass spectrometry have been applied to detect and characterise the metabolites. The metabolite formed by oxidative cleavage of the chain at the carbon atom adjacent to the piperazine nitrogen has been identified as the major plasma metabolite. Glucuronide conjugate of hydroxy-RGH-1756 has been found as one of the main metabolites excreted in the bile where the unchanged compound has not been detected.

Effect of allylestrenol exposure during pregnancy on the activity of microsomal enzyme system (PSMO) of rat in the F1 and F2 generations.

Treatment of pregnant rats with allyestrenol on the 15th and 19th days of pregnancy induced the activity of enzyme system of adult offspring in females while it did not influence it in males. The steroid anticoncipient treatment of adults decreased the activity of some enzymes of the PSMO. The adulthood anticoncipient steroid treatment of rats which encountered allylestrenol in their fetal age compared to the only anticoncipient steroid treated group produces an increased activity of the majority of the tested parameters both in females and males. The effect of the adulthood alcohol treatment was not influenced by the embryonic treatment with allylestrenol. Differences in the second generation are non-significant.

Administration of benzpyrene and allylestrenol in fetal or neonatal periods of life: does it make difference in the microsomal activity?

Administration of allylestrenol or benzpyrene in fetal and neonatal periods of life make the rats susceptible to phenobarbiturate induction. Changes to controls could be observed in three of the enzyme activities tested (p-nitrophenol-hydroxylase, cytochrome P450 and aniline-hydroxylase), with a dominance of the p-nitrophenol-hydroxylase among them. There seemed to be no difference in the action of allylestrenol and benzpyrene, although treatment protocol in the neonatal period proved to be more effective.

Modification of DENA-induced hepatocarcinogenesis by CCl4 cirrhosis. Comparison of the marker enzyme patterns.

The modifying action of chronic liver injury on the process of hepatocarcinogenesis was investigated. To induce cirrhosis or fibrosis F344 rats received CCl4 alone or in combination with phenobarbital, either before (model 1) or after (model 2) the application of initiator, diethylnitrosamine (DENA). In these models, morphology, tumor incidence as well as polysubstrate monooxygenase system, gamma-glutamyltransferase (GGT) and glucose-6-phosphatase (G-6-Pase) were studied. The data presented show that in model 1 the tumor incidence was much lower than in rats treated with DENA alone. This reduction appeared to be associated with the decrease in cytochrome P450 content occurring in model 1 after DENA administration. Promotion of the hepatocarcinogenic process was observed when CCl4 injury followed the application of DENA (model 2). Comparison of marker enzymes in cirrhotic livers and in tumors either with or without cirrhosis indicated that changes in cytochrome P450 and G-6-Pase were rather the results of parenchymal damage, while GGT was elevated only in tumorous livers. In tumorous livers none of the xenobiotic metabolizing activities decreased as much as the cytochrome P450 content of the same samples. Thus conceivably the cytochrome P450 operates more rapidly in tumors than in normal livers.

Effect of acute salinomycin-tiamulin toxicity on the lipid peroxide and antioxidant status of broiler chicken.

The combined effect, if any, of salinomycin poisoning and salinomycin-tiamulin interaction on lipid-peroxidative processes and the antioxidative defence system of the liver was studied in domestic fowl. Male broilers (28-day-old), reared on a diet containing 60 mg/kg salinomycin, were treated intraoesophageally with salinomycin (140 mg/kg body mass) or tiamulin (50 mg/kg body mass). Malondialdehyde, reduced glutathione and cytochrome P-450 concentrations as well as glutathione peroxidase and catalase activities of the liver were determined. Liver malondialdehyde concentration rose in the salinomycin-treated group while the amount of cytochrome P-450 increased in both groups treated. Glutathione concentration and glutathione peroxidase activity of the liver decreased rapidly but hepatic catalase activity increased in both groups after the treatment. Manifestation of the effect exerted by salinomycin and salinomycin-tiamulin on lipid-peroxidative processes nearly coincided with the onset of clinical signs and preceded the increase of hepatic cytochrome P-450 concentration. According to the results, the background of the previously reported incompatibility between salinomycin and tiamulin is the synergistic effect exerted on the antioxidant (glutathione) system.

Effect of contraceptive treatment on thymic glucocorticoid receptors and hepatic microsomal enzyme system of adult rats.

Contraceptive steroid treatment accounted for about a 30 per cent decrease in the number of thymic glucocorticoid receptors of adult rats. Neonatal allylestrenol treatment had no influence on that treatment. The activity of the hepatic microsomal (PSMO) enzyme system was not changed by the contraceptive treatment. It appears that contraceptive treatment may account for overlaps on receptors in adulthood.

Persistent influence of neonatal 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) treatment on glucocorticoid receptors and on the microsomal enzyme system.

Neonatal treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) accounted for a considerable decrease in the number of thymic glucocorticoid receptors in both male and female rats, as assessed at 6 weeks of age. TCDD also gave rise to a marked and prolonged increase in microsomal enzyme activity in the female rats but had practically no such effect on the males. These experimental observations attract attention on to the lasting microsomal inducer effect of the herbicide contaminant dioxin which damages foreign receptors and substantiate the chemical imprinting potential of aromatic hydrocarbons.

Lasting impact of a single benzpyrene treatment in pre-natal and growing age on the thymic glucocorticoid receptors of rats.

1. Rats exposed to benzpyrene in utero at 19 days of pre-natal life showed a relative decrease in the number of thymic glucocorticoid receptors at 6 weeks of age. 2. Primary exposure to benzpyrene at 6 weeks of age had a similar effect on females 4 weeks later, but did not change the glucocorticoid receptor number of males. 3. In utero exposure accounted for an increase in the fetal cytochrome P450 level within 1 day, whereas exposure at 6 weeks of age did not change it within 4 weeks. 4. It appears that exposure to benzpyrene gives rise to a faulty imprinting of the thymic glucocorticoid receptor in both fetal and growing age, to judge from a lasting change in the receptor number.

Impact of neonatal benzpyrene imprinting on liver microsomal enzyme induction by benzpyrene and phenobarbital in adulthood.

A single neonatal treatment of rats with benzpyrene accounted for a durable induction of the liver microsomal enzyme (PSMO) system. Neonatal treatment with benzpyrene enhanced the inducing action of phenobarbital administrated in adulthood, but did not change the effect of benzpyrene treatment in the adult age. It follows that the imprinting and inducing effects of the neonatal benzpyrene treatment took different trends.

Duration of the microsomal enzyme (cytochrome b560ms) inducer action and imprinting potential of a steroid in Tetrahymena.

Tetrahymena subjected to a three-day exposure to the steroid triamcinolone exhibited a decreased microsomal enzyme activity, three days after treatment, followed by a slight and a considerable activity increase over the control at one and two weeks after treatment, respectively. Reexposure to triamicinolone after return to plain medium for three days accounted for a marked increase in the inducer action which, however, failed to persist for an appreciable time.

Effect of the absence of neonatal testosterone imprinting on the activity of the microsomal enzyme system and on the dexamethasone binding of the thymus in adulthood.

Healthy and neonatally castrated male rats were treated with testosterone twice perinatally, while other groups were treated with testosterone also in adulthood or received testosterone only in adulthood. Castration resulted in a moderate (but in some instances significant) decrease of PSMO (polysubstrate monooxygenase) level measured in adulthood. The decrease could partially be compensated by perinatal testosterone treatment. Further testosterone treatment administered in adulthood did not result in further alteration when compared either with the controls or with the neonatally treated animals. However, since in the controls the second testosterone treatment (following the neonatal one), had a decreasing effect, therefore the testosterone treatment administered in adulthood was responsible for the disappearance of the difference between the castrated animals and the controls treated both perinatally and in adulthood. On the basis of these findings it seems likely that the perinatal presence of testosterone plays a major role in the development of enzymatic imprinting and thus, in securing the capability of the liver to split testosterone in adulthood. Since testosterone influences the glycocorticoid receptors of the thymus (presumably by its overlapping effect), so the amount of free glycocorticoid receptors is always higher in the animals castrated neonatally than in the controls. Conversely, neonatal testosterone treatment somewhat increases the number of receptors detectable in adulthood.

Hormonal imprinting of the microsomal enzyme system in adults. Microsomal activity change in response to estrogen (DES, AE) treatment during liver regeneration.

Estrogen (diethylstilbestrol-DES or allylestrenol-AE) treatment applied to rats of both sexes during liver regeneration following subtotal hepatectomy had a long lasting influence on the inducibility by phenobarbital of the hepatic microsomal enzyme system of the females. The enzyme activities of the DES-treated females differed hardly from the baseline two weeks after treatment, but increased almost two-fold over control on induction with phenobarbital 5 and 7 weeks later. The AE-treated females showed a smaller although yet significant, enzyme activity increase only at 7 weeks. The influence of estrogens was negligible, and inhibitory rather than stimulatory, in the males. It appears that, in appropriate conditions, enzyme imprinting can also be induced in adult organisms, since, in all probability, availability for imprinting depends not so much on the age of the organism, as on the developmental state of the target cell.

A single neonatal treatment with methylcholanthrene or benzo(a)pyrene alters microsomal enzyme activity for life.

A single neonatal exposure to methylcholanthrene or benzo(a)-pyrene altered the hepatic microsomal enzyme activity of rats for a long time. Methylcholanthrene depressed, whereas benzo(a)pyrene enhanced the activity of the microsomal enzyme system. The former had a greater influence on males than on females, whereas the latter acted practically uniformly on both sexes.

Influence of single neonatal treatment with allylestrenol or diethylstilbestrol on microsomal enzyme activity of rat liver in adulthood.

A single neonatal treatment of rats with a steroid (allylestrenol or diethylstilbestrol) did not alter the later activity of the hepatic microsomal (cytochrome P-450) enzyme system, but inhibited the inducer action of another steroid (testosterone) administered at the age of six weeks. This suggests that a hormonal imprinting-like mechanism also operates in the case of enzymes.

Inducibility of the hepatic microsomal monooxygenase system experimentally reduced to a minimum amount/activity. Effect of hypophysectomy, partial hepatectomy and phenobarbital/toluene treatment on the hepatic polysubstrate monooxygenase system in rats.

Sham-operated, hypophysectomized, partially hepatectomized and hypophysectomized + partially hepatectomized groups of CFY rats were treated with phenobarbital (40 mg/kg/day) per os or physiological saline once a day, or toluene-vapour (inhalation: 4,000 mg/m3, 8 h/day), for 3 days. The polysubstrate monooxygenase (PSMO) system of the liver was induced by phenobarbital after all kinds of surgical interventions. Inducibility of the enzyme system was the highest in the group with combined hypophysectomy + partial hepatectomy and decreased in order in the groups with partial hepatectomy, hypophysectomy and sham-operation. The relative cytochrome P-450 content (quantity of cytochrome P-450/100 g b.w.) was the lowest after combined operation, higher in partially hepatectomized and hypophysectomized animals and it was the highest in the sham-operated group. Hypophysectomy after partial hepatectomy seems to inhibit hepatic regeneration and to increase the inducibility of the enzyme by phenobarbital, at the same time. Phenobarbital treatment brought about SER proliferation in a part of liver cells, in each group. In case of repeated liver damage due to the consequences of hypophysectomy, partial hepatectomy, phenobarbital administration, a group of the liver cells responds to the first, another to the second, but even after the third injury a group of the liver cells maintained its regular structure. The hypothetical hepatotoxic effect of toluene, a widely used industrial solvent, has been tested. Hepatotoxicity has been excluded. The minimum non-specific hepatotoxic effect of the solvent was not augmented by either partial hepatectomy or hypophysectomy, or by the combination of the two. In all the experimental groups toluene increased the hepatic cytochrome P-450 level and induced a moderate increase in SER. Inducibility of the enzyme system after toluene inhalation was similar to that of the enzyme system after phenobarbital treatment in each group.

Effect of radio-detoxified endotoxin on the liver microsomal drug metabolizing enzyme system in rats.

E. coli endotoxin (LPS) depresses the hepatic microsomal mono-oxygenase activity. Radio-detoxified LPS (TOLERIN: 60 Co irradiated endotoxin preparation) decreases this biotransforming activity to a smaller extent. Phenobarbital, an inducer of this mono-oxygenase system, failed to induce in LPS-treated animals. In radio-detoxified LPS-treated rats, phenobarbital induced the mono-oxygenase and almost fully restored the biotransformation.

Enzyme inducer effects after liver denervation in the rat.

The part of liver innervation was studied in the process of microsomal enzyme induction elicited by phenobarbital or muscular exercise. Enzyme induction was seen to develop: hexobarbital sleeping times became shorter after partial (vagotomy, coeliectomy) as well as total liver denervation in the rat. Though the present results did not preclude subtile differences, they demonstrated that the microsomal enzyme systems of the liver could be activated in the absence of innervation. Upper median laparatomy, which was used as a sham operation, had the strange effect of elongating the sleeping time. This observation is a warning of a changed rate of drug elimination after explorative laparatomy.