Interactions of hepatic cytochromes P-450 with steroid hormones: Regioselectivity and stereospecificity of steroid metabolism and hormonal regulation of rat P-450 enzyme expression☆

Aflatoxin B₁ (AFB₁) is the most toxic naturally occurring mycotoxin produced by Aspergillus flavus and Aspergillus parasiticus, has been regarded as a critical food contaminant in aquafeed. Furthermore, previous studies indicated that dietary AFB₁ exposure could cause liver tissues injury in mammals and fish. However, the mechanism of dietary AFB₁-induced liver injury of fish has not been fully elucidated. The aim of this study was to investigate the effects of dietary AFB₁ exposure on the liver of northern snakehead (Channa argus) (7.52 ± 0.02 g) through hematological parameters, liver histopathology, and ultrastructural analysis and revealed the possible mechanism of AFB₁-induced northern snakehead liver injury through RNA-Seq analysis. The results of serum biochemical indicators demonstrated that the aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP) as the biomarkers of liver damage, increased significantly at AFB₁ exposure concentrations above 200 μg/kg. Liver histopathology and ultrastructural analysis demonstrated that different degrees of lesions were found in 100, 200, and 400 μg/kg groups. Liver transcriptomic analyses demonstrated that a total of 1307 differentially expressed genes (DEGs) were significantly expressed induced in response to the dietary AFB₁ exposure, of which 540 were up- and 767 were down-regulated genes. Furthermore, Gene Ontology (GO) analysis showed that the ribosome biogenesis, ribonucleoprotein complex biogenesis, negative regulation of cellular process, and metabolic process were highly enriched in the biological process category. Meanwhile, 310 enriched pathways were identified in the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway database analysis, of which the metabolism of xenobiotics by cytochrome P450, glutathione metabolism, protein processing in endoplasmic reticulum, and PI3K-Akt signaling pathway were the vital metabolic mechanisms related to dietary AFB₁ exposure-induced liver injury in northern snakehead and were confirmed by qPCR. Overall, the findings provide more valuable information for further studies on the latent toxicity mechanisms of AFB₁ in fish.

The purpose of this study was to investigate the effects of dietary aflatoxin B₁ (AFB₁) on growth performance and AFB₁ biotransformation, and hepatic oxidative stress, endoplasmic reticulum (ER) stress, apoptosis, and inflammation in northern snakehead (Channa argus). A total of 600 northern snakeheads (7.52 ± 0.02 g) were divided into five groups (three replicates/group) and fed the diets with AFB₁ at concentrations of 0, 50, 100, 200, and 400 ppb for 8 weeks. The results demonstrated that dietary AFB₁ (≥ 200 ppb) reduced FBW, WG, and SGR. 100, 200, and 400 ppb AFB₁ treatment groups significantly decreased the PER, CRP, C3, C4, IgM, and LYS levels in northern snakehead, while FCR was significant increased. Moreover, dietary AFB₁ (100, 200, and 400 ppb) increased cyp1a, cyp1b (except 400 ppb), and cyp3a mRNA expression levels, while reducing the GST enzymatic activity and mRNA expression levels in northern snakehead. Furthermore, AFB₁ (≥ 100 ppb) increased ROS, MDA, and 8-OHdG levels, and grp78, ire1, perk, jnk, chop, and traf2 mRNA expression levels, and decreased SOD, CAT, GSH-Px, and GSH (except 100 ppb) levels and the gene expression levels of cat, gsh-px (except 100 ppb), and Cu/Zn sod. In addition, AFB₁ (100, 200, and 400 ppb) up-regulated the cyt-c, bax, cas-3, and cas-9 mRNA levels in the liver, while down-regulating the bcl-2 expression levels. Meanwhile, the expression levels of nf-κb, tnf-α (except 100 ppb), il-1β, and il-8 in the liver were up-regulated in AFB₁ treatment groups (≥ 100 ppb), while the iκbα mRNA levels were down-regulated. In summary, dietary AFB₁ reduced growth performance and humoral immunity in northern snakehead. Meanwhile, the cyclic occurrence of oxidative stress and ER stress, and induced apoptosis and inflammation, is one of the main reasons for AFB₁-induced liver injury in the northern snakehead, which will provide valuable information and a fresh perspective for further research into AFB₁-induced liver injury in fish.

Cytochrome P450 (CYP) plays an important role in psychopharmacology. While liver CYP enzymes are responsible for the biotransformation of psychotropic drugs, brain CYP enzymes are involved in the local metabolism of these drugs and endogenous neuroactive substances, such as neurosteroids, and in alternative pathways of neurotransmitter biosynthesis including dopamine and serotonin. Recent studies have revealed a relation between the brain nervous system and cytochrome P450, indicating that CYP enzymes metabolize endogenous neuroactive substances in the brain, while the brain nervous system is engaged in the central neuroendocrine and neuroimmune regulation of cytochrome P450 in the liver. Therefore, the effect of neuroactive drugs on cytochrome P450 should be investigated not only in vitro, but also at in vivo conditions, since only in vivo all mechanisms of drug-enzyme interaction can be observed, including neuroendocrine and neuroimmune modulation.

Psychotropic drugs can potentially affect cytochrome P450 via a number of mechanisms operating at the level of the nervous, hormonal and immune systems, and the liver. Their effect on cytochrome P450 in the brain is often different than in the liver and region-dependent. Since psychotropic drugs can affect cytochrome P450 both in the liver and brain, they can modify their own pharmacological effect at both pharmacokinetic and pharmacodynamic level. The article describes the mechanisms by which psychotropic drugs can change the expression/activity of cytochrome P450 in the liver and brain, and discusses the significance of those mechanisms for drug action and drug-drug interactions. Moreover, the brain CYP2D6 is considered as a potential target for psychotropics.

Toxicology studies conducted with oil-soluble rosemary extracts to support authorization as a food additive (antioxidant) in the EU include an Ames test using a supercritical carbon dioxide extract (D74), a full 90-day study using D74 and an acetone extract (F62), and an investigative 90-day study with a 28-day recovery period (using D74 only). D74 was non-mutagenic in the Ames test. In the full 90-day study, where rats (20/sex/group) were either provided control diet or diets containing D74 (300, 600, or 2400 mg/kg) or F62 (3800 mg/kg), liver enlargement and hepatocellular hypertrophy were observed. To determine a mode of action and assess the reversibility of the hepatic effects, an investigative 90-day study was conducted using female rats (10/group receiving control diet or diet containing 2400 mg/kg D74). Liver enlargement was fully reversible after 28 days and microsomal enzyme analysis revealed reversible induction of cytochrome P450 enzymes (CYP2A1, CYP2A2, CYP2C11, CYP2E1, and CYP4A), demonstrating that the hepatic effects were adaptive and of no toxicological concern. Therefore, the highest dietary concentrations were established as the NOAELs. The investigative 90-day study NOAEL (providing 64 mg/kg bw/day carnosol and carnosic acid [the primary antioxidant components]) was used to establish a temporary ADI for rosemary extracts.

Advances in technology have allowed for the sensitive, specific, and simultaneous quantitative profiling of steroid precursors, bioactive steroids and inactive metabolites, facilitating comprehensive characterization of the serum and urine steroid metabolomes. The quantification of steroid panels is therefore gaining favor over quantification of single marker metabolites in the clinical and research laboratories. However, although the biochemical pathways for the biosynthesis and metabolism of steroid hormones are now well defined, a gulf still exists between this knowledge and its application to the measured steroid profiles. In this review, we present an overview of steroid hormone biosynthesis and metabolism by the liver and peripheral tissues, specifically highlighting the pathways linking and differentiating the serum and urine steroid metabolomes. A brief overview of the methodology used in steroid profiling is also provided.

Obesity is an endemic problem in the United States and elsewhere, and data indicate that in addition to overconsumption, exposure to specific chemicals enhances obesity. CYP2B metabolizes multiple endo- and xenobiotics, and recent data suggests that repression of Cyp2b activity increases dyslipidemia and age-onset obesity, especially in males. To investigate the role played by Cyp2b in lipid homeostasis and obesity, we treated wildtype and Cyp2b-null mice with a normal (ND) or 60% high-fat diet (HFD) for 10 weeks and determined metabolic and molecular changes. Male HFD-fed Cyp2b-null mice weigh 15% more than HFD-fed wildtype mice, primarily due to an increase in white adipose tissue (WAT); however, Cyp2b-null female mice did not demonstrate greater body mass or WAT. Serum parameters indicate increased ketosis, leptin and cholesterol in HFD-fed Cyp2b-null male mice compared to HFD-fed wildtype mice. Liver triglycerides and liver:serum triglyceride ratios were higher than their similarly treated wildtype counterparts in Cyp2b-null male mice, indicating a role for Cyp2b in fatty acid metabolism regardless of diet. Furthermore, RNAseq demonstrates that hepatic gene expression in ND-fed Cyp2b-null male mice is similar to HFD-fed WT male mice, suggestive of fatty liver disease progression and a role for Cyp2b in lipid homeostasis. Females did not show as demonstrative changes in liver health, and significantly fewer changes in gene expression, as well as gene expression associated with liver disease. Overall our data indicates that the repression or inhibition of CYP2B may exacerbate metabolic disorders and cause obesity by perturbing fatty acid metabolism, especially in males.