In vivo effect of dried chicory root (Cichorium intybus L.) on xenobiotica metabolising cytochrome P450 enzymes in porcine liver

Cytochrome P450 (CYP) enzymes are widely studied for their involvement in metabolism of drugs and endogenous compounds. In porcine liver, CYP1A2,2Aand 2E1 are important for the metabolism of skatole.Feeding chicory roots to pigs is known to decrease the skatole concentration in plasma and fat. In the present study we investigated the effect of chicory on CYP mRNA and protein expression, as well as their activity. Male pigs were feed dried chicory root for 16 days before liver samples were collected. By the use of RT-PCR and Western blotting we showed that the mRNA and protein expression of CYP1A2 and 2A were increased in chicory fed pigs. The mRNA expression of CYP2E1 was increased, while there was no effect on protein expression. Activity of CYP1A2 and 2A were increased in chicory feed pigs; this was not the case for CYP2E1 activity. In conclusion; oral administration of chicory root for 16 days to pigs increased the mRNA expression of CYP1A2, 2A and 2E1; and the protein expression of CYP1A2 and 2A. The activities of CYP1A2 and 2A were increased

Regulation of cytochrome P450 mRNA expression in primary porcine hepatocytes by selected secondary plant metabolites from chicory (Cichorium intybus L.)

Chicory (Cichorium intybus) has been shown to induce enzymes of pharmacokinetic relevance (cytochrome P450; CYP). The aim of this study was to investigate the effects of selected secondary plant metabolites with a global extract of chicory root, on the expression of hepatic CYP mRNA (1A2, 2A19, 2C33, 2D25, 2E1 and 3A29), using primary porcine hepatocytes. Of the tested secondary plant metabolites, artemisinin, scoparone, lactucin and esculetin all induced increased expression of specific CYPs, while esculin showed no effect. In contrast, a global extract of chicory root decreased the expression of CYP1A2, 2C33, 2D25 and 3A29 at high concentrations. The results suggest that purified secondary metabolites from chicory affect CYP expression and thereby might affect detoxification in general, and that global extracts of plants can have effects different from individual components

Feeding driet chicory root to pigs decrease androstenone accumulation in fat by increasing hepatic 3β hydroxysteroid dehydrogenase expression

The present study investigated the in vivo effect of chicory root on testicular steroid concentrations and androstenone metabolizing enzymes in entire male pigs. Furthermore, the effect on skatole and indole concentrations in plasma and adipose tissue was investigated. The pigs were divided into two groups; one receiving experimental feed containing 10% dried chicory root for 16 days before slaughter, the control group was fed a standard diet. Plasma, adipose and liver tissue samples were collected at slaughter. Plasma was analyzed for the concentration of testosterone, estradiol, insulin-like growth factor 1 (IGF-1), skatole and indole. Adipose tissue was analyzed for the concentration of androstenone, skatole and indole, while the liver tissue was analyzed for mRNA and protein expressions of 3β-hydroxysteroid dehydrogenase (3β-HSD), sulfotransferase 2A1 and heat-shock protein 70 (HSP70). The results showed that the androstenone concentrations in the adipose tissue of chicory fed pigs were significantly (p < 0.05) lower and indole concentrations were higher (p < 0.05) compared to control fed pigs. Moreover the chicory root fed pigs had increased mRNA and protein expression of 3β-HSD and decreased HSP70 expression (p < 0.05). Testosterone and IGF-1 concentrations in plasma as well as skatole concentrations in adipose tissue were not altered by dietary intake of chicory root. It is concluded that chicory root in the dietreduces the concentration of androstenone in adipose tissue via induction of 3�-HSD, and that these changes were not due to increased cellular stress

Regulation of Porcine Hepatic Cytochrome P450 by Chicory Root - Implication of Boar Taint

Dietary components have been shown to affect the hepatic detoxification system. Hepatic clearance of xenobiotics is performed in two steps, Phase I and II. Phase I is usually carried out by enzymes belonging to the cytochrome P450 (CYP) family, while Phase II is carried out by a more diverse group. Phase I and II enzymes are extensively studied for their importance in drug clearance and in pigs for their impact on the occurrence of boar taint. It is generally believed that sufficient Phase I and II metabolism of the boar taint compounds, skatole and androstenone, diminish the occurrence of tainted meat from sexually mature male pigs. Moreover, dietary chicory root has been shown to decrease the occurrence of boar taint. The present PhD study was carried out order to investigate the regulation of Phase I and II enzymes by chicory root. Sexually mature male pigs were fed chicory root for the ast 16 days before slaughter. They showed increased mRNA expression of CYP1A2, 2A19, 2D25, 2E1, 3A29 as well as 3β hydroxysteroid dehydrogenase (3β-HSD). A simultaneous increase in protein expression of CYP1A2, 2A19 and 3β-HSD as well as increased activities of CYP1A2, 2A and 3A were shown. No effect was shown on the expression of the investigated Phase II enzyme, sulfotransferase 2A1 (SULT2A1). Chicory feeding had no effect on skatole accumulation in fat, while the androstenone concentration found in the group of chicory fed pigs were lower than in the control group. Additionally, chicory feeding had no effect on testosterone, estradiol and insulin-like growth factor 1 (IGF-1) in plasma. The obtained in vivo results of chicory feeding are in contrast to the results obtained in vitro using primary porcine hepatocytes. Porcine hepatocytes were isolated from female piglets and treated with a methanolic extract of chicory root for 24 hours. This treatment down regulated the mRNA expression of CYP1A2, 2C33, 2D25 and 3A29. In comparison standard CYP inducers (β-naphthoflavone and dexamethasone) and secondary plant metabolites (artemisinin and scoparone) increased the mRNA expression of the same CYPs. As the occurrence of boar taint is associated with sexually mature male pigs, a comparison of the constitutive CYP expression and activity between genders was also included. It was shown that female pigs, in comparison to entire male pigs, had greater mRNA expression of CYP1A2, 2A19 and 3A29, while no differences were observed in protein expression. Likewise female pigs had greater CYP1A2, 2A and 2C enzyme activity. The absolute activities found in the chicory fed male pigs were comparable to the activities found in the female pigs. In conclusion, the results presented in this PhD thesis uggest that chicory feeding can be used to lower the occurrence of boar taint. This is partly based on the finding that chicory feeding increases CYP activity to levels comparable with female pigs, as well as the effects shown on the metabolism of androstenone

7-Hydroxylation of warfarin is strongly inhibited by sesamin, but not by episesamin, caffeic and ferulic acids in human hepatic microsomes

© 2018 Elsevier Ltd Warfarin is a commonly used anticoagulant drug and is a derivate of coumarin. Cytochrome P450 2C9 (CYP2C9) plays the key role in transformation of coumarin and thus, influences determination of warfarin dosage. A number of factors including dietary compounds such as sesamin, caffeic acid and ferulic acids can regulate the activity of CYP2C9. The present study tested the hypothesis that sesamin, episesamin, caffeic acid and ferulic acid decreases the rate of warfarin 7-hydroxylation via inhibition of hepatic CYP2C9. The experiments were conducted on hepatic microsomes from human donors. It was demonstrated that the rate of 7-hydroxylation of warfarin was significantly decreased in the presence of sesamin in the range of concentrations from 5 to 500 nM, and was not affected by episesamin, caffeic acid and ferulic acid in the same range of concentrations. The kinetic analysis indicated non-competitive type of inhibition by sesamin with Ki = 202 ± 18 nM. In conclusion, the results of our in vitro study revealed that sesamin was able to inhibit formation of a major metabolite of warfarin, 7-hydroxywarfarin. The potentially negative consequences of the consumption of high amounts of sesamin-containing food or dietary supplements in warfarin-treated patients need to be further studied

Naringenin and falcarinol stimulate glucose uptake and TBC1D1 phosphorylation in porcine myotube cultures

Abstract: Insulin resistance in muscles is a major problem associated with Type 2 diabetes. Bioactive compounds of plant origin have long been known for possessing anti-diabetic properties. We have studied the effect of the bioactive compounds naringenin (dihydroflavonol) and falcarinol (polyacetylene) on glucose uptake (GU) in normal and insulin resistant primary porcine myotubes, in the presence and absence of insulin to identify signaling pathways mediating their effects on GU. The dependence on glucose transporter type 4 (Glut4) activity, insulin signaling and AMP-activated protein kinase (AMPK)-signaling was studied by using the Glut4 inhibitor indinavir, the phosphatidyl inositol-3 kinase (PI3K) and p38 mitogen activated protein kinase (MAPK) inhibitor wortmannin, and the AMPK inhibitor dorsomorphin (DM), respectively. Naringenin and falcarinol stimulated GU was attenuated in the presence of indinavir and wortmannin, indicating a dependence on Glut4 activity as well as PI3K and/or p38MAPK activity. By contrast, DM diminished GU induced by naringenin only, indicating that falcarinol-stimulated GU was independent of AMPK activity. Finally, we show that naringenin and falcarinol enhance phosphorylation of TBC1D1 suggesting that these compounds enhance translocation of Glut4 containing vesicles and thereby GU via a TBC1D1-dependent mechanism

Effects of Multi-Component Mixtures from Sewage Treatment Plant Effluent on Common Carp (Cyprinus carpio) under Fully Realistic Condition.

This study characterized changes in biomarker responses in common carp (Cyprinus carpio) upon exposure to effluent water discharged from a sewage treatment plant (STP) under real conditions. Fish were exposed to contamination in Cezarka pond, which receives all of its water input from the STP in the town of Vodnany, Czech Republic. Five sampling events were performed at day 0, 30, 90, 180, and 360 starting in April 2015. In total, 62 pharmaceutical and personal care products (PPCPs) were detected in the polar organic chemical integrative sampler. Compared to a control pond, the total concentration of PPCPs was 45, 16, 7, and 7 times higher in Cezarka pond at day 30, 90, 180, and 360, respectively. The result of oxidative stress and antioxidant enzyme biomarkers indicated alterations in the liver and intestine tissues of fish from Cezarka pond at day 30 and 360, respectively. High plasma vitellogenin levels were observed in both exposed females (180 and 360 days) and males (360 days) compared with their respective controls. However, only exposed female fish had higher vitellogenin mRNA expression than the control fish in these periods. Exposed female fish showed irregular structure of the ovary with scattered oocytes, which further developed to a vitellogenic stage at day 360. Low white blood cell levels were indicated in all exposed fish. Despite numerous alterations in exposed fish, favorable ecological conditions including high availability of food resulted in a better overall condition of the exposed fish after 1 year of exposure compared to the controls

Immunocastration of male pigs – situation today

Immunological castration of male pigs is an attractive alternative to surgical castration and nowadays is increasingly used in many countries to reduce boar taint and improve pork quality. Moreover, immunocastrated pigs showed reduced sexual and aggressive behaviour compared to entire male pigs, thus improving animal welfare. Carcass and meat quality parameters generally do not differ between immunocastrated and surgically castrated male pigs. (C) 2015 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license

Regulatio of 3β-hydroxysteroid dehydrogenase and sulphotransferase 2A1 gene expression in primary porcine hepatocytes by selected sex-steroids and plant secondary metabolites from chicory (Cichorium intybus L.) and wormwood (Artemisia sp.)

In pigs the endogenously produced compound androstenone is metabolised in the liver in two steps by 3β-hydroxysteroid dehydrogenase (3β-HSD) and sulphotransferase 2A1 (SULT2A1). The present study investigated the effect of selected sex-steroids (0.01–1 μM androstenone, testosterone and estradiol), skatole (1-100 μM) and secondary plant metabolites (1–100 μM) on the expression of 3β-HSD and SULT2A1 mRNA. Additionally the effect of a global methanolic extract of dried chicory root was investigated and compared to previous obtained in vivo effects. Primary hepatocytes were isolated from the livers of piglets (crossbreed: Landrace × Yorkshire and Duroc) and cultured for 24 h before treatment for an additionally 24 h. RNA was isolated from the hepatocytes and specific gene expression determined by RT-PCR using TaqMan probes. The investigated sex-steroids had no effect on the mRNA expression of 3β-HSD and SULT2A1, while skatole decreased the content of SULT2A1 30% compared to control. Of the investigated secondary plant metabolites artemisinin and scoparone (found in Artemisia sp.) lowered the content of SULT2A1 by 20 and 30% compared to control, respectively. Moreover, we tested three secondary plant metabolites (lactucin, esculetin and esculin) found in chicory root. Lactucin increased the mRNA content of both 3β-HSD and SULT2A1 by 200% compared to control. An extract of chicory root was shown to decrease the expression of both 3β-HSD and SULT2A1. It is concluded that the gene expression of enzymes with importance for androstenone metabolism is regulated by secondary plant metabolites in a complex manner