title: Regulation 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.).
abstract: 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 24h before treatment for an additionally 24h. 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.
authors: Department of Food Science, Aarhus University, Denmark. Electronic address: martink.rasmussen@agrsci.dk.
authors: Martin Krøyer
authors: MK
authors: Rasmussen
authors: Department of Food Science, Aarhus University, Denmark.
authors: Bo
authors: B
authors: Ekstrand
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000235:genetics*; D000818:Animals; D019306:Artemisia / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D018651:Cichorium intybus / Q000737:chemistry / Q000378:metabolism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D012739:Gonadal Steroid Hormones / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D010936:Plant Extracts / Q000378:metabolism / Q000494:pharmacology*; D061251:Primary Cell Culture; D064210:Secondary Metabolism; D015238:Sulfotransferases / Q000235:genetics*; D013552:Swine
chemical_list: D012739:Gonadal Steroid Hormones; D010936:Plant Extracts; D015067:17-Hydroxysteroid Dehydrogenases; C044256:3 (or 17)-beta-hydroxysteroid dehydrogenase; D015238:Sulfotransferases; C028152:alcohol sulfotransferase
keywords: 3β Hydroxysteroid dehydrogenase; 3β-HSD; 4-(2-Hydroxyethyl)piperazine-1-ethanesulphonic acid; ANOVA; AhR; BSA; CAR; Ct; DHEA; DMSO; DPBS; Dulbecco's phosphate buffered saline; EGTA; FCS; GAPDH; Gene regulation; HEPES; PCR; PXR; Primary porcine hepatocytes; RT-PCR; SEM; SQL; SULT2A1; Secondary plant metabolites; Sex-steroids; WME; Williams E medium; XR; analysis of variance; aryl hydrocarbon receptor; bovine serum albumin; cDNA; complementary DNA; constitutive androstane receptor; dehydroepiandrosterone; dimethyl sulphoxide; ethylene glycol tetraacetic acid; foetal calf serum; glyceraldehyde 3-phosphate dehydrogenase; mRNA; messenger RNA; pregnane X receptor; reverse transcriptase polymerase chain reaction; sesquiterpene lactone; standard error of the mean; sulphotransferase 2A1; threshold cycle; xenobiotic receptor