CYP2E1 Research Articles

Relationship between Cytochrome P450 2E1 Gene Polymorphism and Susceptibility of Low Birth Weight Infants

ABSTRACT The purpose of this study is to investigate the relationship between cytochrome P450 2E1 (CYP2E1) RsaI/PstI and DraI single gene polymorphism (SNP) and low birth weight infants (LBW) of Chinese population. The researchers detected CYP2E1 RsaI/PstI and DraI SNPs by real-time PCR cycling and HRM analysis in 461 premature delivery and 461 healthy pregnant women. This paper found that RsaI/PstI C2C2 was statistically significantly correlation with increased risk of LBW (Odds ratios= 0.66,95% CI: 0.44–0.98, P = 0.04) and (Odds ratios = 0.75 , 95% CI: 0.56–0.99, P=0.04) for C2C2, compared with C1C1 and C1C1 + C1C2, respectively. When CYP2E1 RsaI/PstI and DraI SNPs combine, the risk of low birth weight infants was increased in individuals with both DraI TT and RsaI/ PstI C2C2 genotypes (Odds ratios=0.59, 95% CI=0.36–0.97). The current study demonstrates CYP2E1 RsaI/PstI polymorphism is significantly related to LBW and the interaction between the RsaI/PstI and DraI polymorphisms has a significant impact on LBW.

Ameliorative impacts of chrysin against gibberellic acid-induced liver and kidney damage through the regulation of antioxidants, oxidative stress, inflammatory cytokines, and apoptosis biomarkers.

Gibberellic acid (GA3), a widely known plant growth regulator, has been mostly used in agriculture. Little is known regarding its toxicity or the impact of its metabolic mechanism on human health. The current study examined the protective impact of chrysin against GA3-induced liver and kidney dysfunctions at biochemical, molecular, and histopathological levels. Forty male albino rats were allocated into 4 groups. The control group received saline; the chrysin group received 50mg/kg/BW orally daily for 4 weeks; the GA3 group received 55mg/kg/BW GA3 via daily oral gavage for 4 weeks, and the protective group (chrysin + GA3) was administered both chrysin and GA3 at the same dosage given in chrysin and GA3 groups. Chrysin was administered 1 h earlier than GA3. The GA3 induced liver and kidney injuries as proven by the elevation of hepatic and renal markers with a significant increase in malondialdehyde levels. Furthermore, a decrease of catalase and glutathione was reported in the GA3-administered rats. Pre-administration of chrysin significantly protected the hepatorenal tissue against the deleterious effects of GA3. Chrysin restored the hepatorenal functions and their antioxidant ability to normal levels. Moreover, chrysin modulated the hepatorenal toxic effects of GA3 at the molecular level via the upregulation of the antiapoptotic genes, interleukin-10 (IL-10), hemoxygenase-1, and nuclear factor erythroid 2-related factor 2 expressions; the downregulation of the kidney injury molecule-1 and caspase-3 mRNA expressions; and a decrease in IL-1β and tumor necrosis factor-α secretions. Additionally, the pre-administration of chrysin effectively attenuated the GA3-induced hepatorenal histopathological changes by regulating the immunoexpression of cytochrome P450 2E1 (CYP2E1) and pregnane X receptor, resulting in normal values at the cellular level. In conclusion, chrysin attenuated GA3-induced oxidative hepatorenal injury by inhibiting free-radical production and cytokine expression as well as by modulating the antioxidant, apoptotic, and antiapoptotic activities. Chrysin is a potent hepatorenal protective agent to antagonize oxidative stress induced by GA3.

Orientin reverses acetaminophen-induced acute liver failure by inhibiting oxidative stress and mitochondrial dysfunction

Oxidative stress, as an important pathogenic factor, plays a critical role in acetaminophen (APAP) overdose-induced acute liver failure (ALF). Thus, an antioxidative strategy may be a good way to alleviate APAP-induced liver damage. Previous research has reported that Orientin (Ori) possesses antioxidant, anti-inflammatory and anticancer effects. This study aimed to explore whether Ori can protect against APAP-induced oxidative stress and to elucidate its underlying mechanism. Our results indicated that Ori alleviated APAP-induced hepatic pathological changes by reducing mouse mortality, inhibiting the expression of cytochrome P450 2E1 (CYP2E1), maintaining a normal liver structure, and reducing the levels of serum alanine transaminase (ALT) and serum aspartate aminotransferase (AST). Moreover, Ori protected against APAP-induced oxidative damage by decreasing the formation of malondialdehyde (MDA) and myeloperoxidase (MPO) and increasing the levels of superoxide dismutase (SOD) and the GSH-to-GSSG ratio. Moreover, Ori regulated APAP-induced hepatocyte apoptosis and mitochondrial dysfunction by inhibiting cytochrome c mitochondrial translocation and c-jun N-terminal kinase phosphorylation, promoting Bcl-2 expression and reducing Bax and caspase-3 cleavage. Furthermore, Ori not only obviously promoted Nrf2 nuclear translocation but also activated the antioxidant-related proteins HO-1, GCLC, GCLM and NQO1. Therefore, Ori prevented APAP-induced hepatocyte oxidative damage and mitochondrial dysfunction via Nrf2-mediated and JNK/cytochrome c/caspase-3 signaling pathways.

Exploring the Interactome of Cytochrome P450 2E1 in Human Liver Microsomes with Chemical Crosslinking Mass Spectrometry.

Aiming to elucidate the system-wide effects of the alcohol-induced increase in the content of cytochrome P450 2E1 (CYP2E1) on drug metabolism, we explored the array of its protein-protein interactions (interactome) in human liver microsomes (HLM) with chemical crosslinking mass spectrometry (CXMS). Our strategy employs membrane incorporation of purified CYP2E1 modified with photoreactive crosslinkers benzophenone-4-maleimide and 4-(N-succinimidylcarboxy)benzophenone. Exposure of bait-incorporated HLM samples to light was followed by isolating the His-tagged bait protein and its crosslinked aggregates on Ni-NTA agarose. Analyzing the individual bands of SDS-PAGE slabs of thereby isolated protein with the toolset of untargeted proteomics, we detected the crosslinked dimeric and trimeric complexes of CYP2E1 with other drug-metabolizing enzymes. Among the most extensively crosslinked partners of CYP2E1 are the cytochromes P450 2A6, 2C8, 3A4, 4A11, and 4F2, UDP-glucuronosyltransferases (UGTs) 1A and 2B, fatty aldehyde dehydrogenase (ALDH3A2), epoxide hydrolase 1 (EPHX1), disulfide oxidase 1α (ERO1L), and ribophorin II (RPN2). These results demonstrate the exploratory power of the proposed CXMS strategy and corroborate the concept of tight functional integration in the human drug-metabolizing ensemble through protein-protein interactions of the constituting enzymes.

Role of Mitochondrial Cytochrome P450 2E1 in Healthy and Diseased Liver.

Cytochrome P450 2E1 (CYP2E1) is pivotal in hepatotoxicity induced by alcohol abuse and different xenobiotics. In this setting, CYP2E1 generates reactive metabolites inducing oxidative stress, mitochondrial dysfunction and cell death. In addition, this enzyme appears to play a role in the progression of obesity-related fatty liver to nonalcoholic steatohepatitis. Indeed, increased CYP2E1 activity in nonalcoholic fatty liver disease (NAFLD) is deemed to induce reactive oxygen species overproduction, which in turn triggers oxidative stress, necroinflammation and fibrosis. In 1997, Avadhani’s group reported for the first time the presence of CYP2E1 in rat liver mitochondria, and subsequent investigations by other groups confirmed that mitochondrial CYP2E1 (mtCYP2E1) could be found in different experimental models. In this review, we first recall the main features of CYP2E1 including its role in the biotransformation of endogenous and exogenous molecules, the regulation of its expression and activity and its involvement in different liver diseases. Then, we present the current knowledge on the physiological role of mtCYP2E1, its contribution to xenobiotic biotransformation as well as the mechanism and regulation of CYP2E1 targeting to mitochondria. Finally, we discuss experimental investigations suggesting that mtCYP2E1 could have a role in alcohol-associated liver disease, xenobiotic-induced hepatotoxicity and NAFLD.

Investigation of CYP2E1 and Caspase-3 Gene Expressions in COVID-19 patients

BackgroundCOVID-19 pandemic spread around the world like an infectious disease that presents waved effects on patients. Some patients needed ICU and respiratory support. Some patients only had flu-like symptoms. Cytokine storm and elevated ROS were serious problems for treatment. Apoptotic genes and CYP Family are part of these mechanisms. AimIn this study, our aim was to examine the gene expression CYP2E1 and Caspase-3 in patients with COVID-19 infection. Method60 COVID-19(+) patients (ICU and non-ICU patients) and 30 healthy volunteers were enrolled to study. To measure the level of gene expression qPCR was used. The 2-ΔΔCt method was utilized to analyze gene expression. ResultsThe expression of CYP2E1 and Caspase-3 genes showed a significant discrepancy between patients and healthy individuals. Caspase-3 expression increased (p=0,0041) but CYP2E1 expression decreased (p=0,0214) in COVID-19 patients compared to healthy individuals. Both levels of gene expression were lower in patients with affected lungs than patients with unaffected lungs (p<0,05). Laboratory findings including d-Dimer, LDH, platelet count, lymphocyte count were related to both gene expressions (p<0,05). We found no correlation between CYP2E1 and Caspase-3 expressions. ConclusionThe expression of Caspase-3 demonstrated apoptotic situations of patients but was not related to the CYP2E1 expression level. CYP2E1 gene expression is an important actor to metabolize endogens and xenobiotics however, COVID-19 patients demonstrated decreased CYP2E1 expression. CYP2E1 and Caspase-3 gene expression levels may be used as a diagnostic tool for COVID-19 patients.

Alcohol induced hepatic retinoid depletion is associated with the induction of multiple retinoid catabolizing cytochrome P450 enzymes.

Chronic alcohol consumption leads to a spectrum of liver disease that is associated with significant global mortality and morbidity. Alcohol is known to deplete hepatic vitamin A content, which has been linked to the pathogenesis of alcoholic liver disease. It has been suggested that induction of Cytochrome P450 2E1 (CYP2E1) contributes to alcohol-induced hepatic vitamin A depletion, but the possible contributions of other retinoid-catabolizing CYPs have not been well studied. The main objective of this study was to better understand alcohol-induced hepatic vitamin A depletion and test the hypothesis that alcohol-induced depletion of hepatic vitamin A is due to CYP-mediated oxidative catabolism. This hypothesis was tested in a mouse model of chronic alcohol consumption, including wild type and Cyp2e1 -/- mice. Our results show that chronic alcohol consumption is associated with decreased levels of hepatic retinol, retinyl esters, and retinoic acid. Moreover, the depletion of hepatic retinoid is associated with the induction of multiple retinoid catabolizing CYPs, including CYP26A1, and CYP26B1 in alcohol fed wild type mice. In Cyp2e1 -/- mice, alcohol-induced retinol decline is blunted but retinyl esters undergo a change in their acyl composition and decline upon alcohol exposure like WT mice. In conclusion, the alcohol induced decline in hepatic vitamin A content is associated with increased expression of multiple retinoid-catabolizing CYPs, including the retinoic acid specific hydroxylases CYP26A1 and CYP26B1.

Diallyl disulfide prevents 1,3-dichloro-2-propanol-induced hepatotoxicity through mitogen-activated protein kinases signaling

We investigated whether diallyl disulfide (DADS) has protective effects against 1,3-dichloro-2-propanol (1,3-DCP)-induced hepatotoxicity and oxidative damage in rats and HepG2 cells. DADS was administered to rats once daily for 7 days at doses of 30 and 60 mg/kg/day. One hour after the final DADS treatment, the rats were administered 90 mg/kg 1,3-DCP to induce acute hepatotoxicity. DADS treatment significantly suppressed the increase in serum aminotransferase levels induced by 1,3-DCP administration, and reduced histopathological alterations in the liver. DADS treatment reduced 1-3-DCP-induced apoptotic changes in the liver, as revealed by terminal deoxynucleotidyl transferase dUTP nick end labeling staining and immunohistochemistry for caspase-3. DADS treatment competitively inhibited or reduced cytochrome p450 2E1 (CYP2E1) expression, which is involved in the metabolic activation of 1,3-DCP, and enhanced antioxidant properties. Furthermore, DADS treatment inhibited phosphorylation of mitogen-activated protein kinases (MAPKs) and apoptotic signaling. In in vitro experiments, MAPKs inhibitors reduced the expression of Bax/Bcl-2/Caspase 3 signaling, which effects were more significant in co-treated cells with DADS and MAPKs inhibitors. In conclusion, the protective effect of DADS against 1,3-DCP-induced hepatotoxicity may be related to blocking the metabolic activation of 1,3-DCP by suppressing CYP2E1 expression, inducing antioxidant enzyme activity, and reducing apoptotic activity by inhibiting phosphorylation of MAPKs.

Extracts of Poria cocos polysaccharides improves alcoholic liver disease in mice via CYP2E1 and NF-κB inflammatory pathways

The present study investigated the effect of extract of Poria cocos polysaccharides(PCP) on cytochrome P450 2 E1(CYP2 E1) and nuclear factor κB(NF-κB) inflammatory signaling pathways in alcoholic liver disease(ALD) mice and explored its protective effect and mechanism. Sixty male C57 BL/6 N mice of SPF grade were randomly divided into a control group, a model group, a positive drug group(bifendate, 200 mg·kg~(-1)), and high-(200 mg·kg~(-1)) and low-dose(50 mg·kg~(-1)) PCP groups. Gao-binge mo-del was induced and the mice in each group were treated correspondingly. Liver morphological and pathological changes were observed and organ index was calculated. Serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected. Malondialdehyde(MDA) and superoxide dismutase(SOD) in liver tissues were detected by assay kits. The levels of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) were detected by ELISA. The activation of macrophages was observed by immunofluorescence staining and protein expression of CYP2 E1, Toll-like receptor 4(TLR4), NF-κB p65, and phosphorylated NF-κB p65(p-NF-κB p65) were analyzed by Western blot. The ALD model was properly induced. Compared with the model group, the PCP groups significantly improved the pathological injury of liver tissues. Immunofluorescence staining revealed that compared with the model group, the groups with drug intervention showed decreased macrophages in liver tissues. Additionally, the PCP groups showed reduced ALT, AST, MDA, IL-6, and TNF-α(P&lt;0.05), and potentiated activity of SOD(P&lt;0.01). PCP extract has the protective effect against alcoholic liver injury in mice, and the underlying mechanism may be related to the regulation of the expression of CYP2 E1 and inhibition of TLR4/NF-κB inflammatory signaling pathway to reduce oxidative stress and inflammatory injury, thereby inhibiting the development of ALD.

The Role and Molecular Regulatory Mechanism of A Disintegrin and Metalloprotease 9 in Alcoholic Liver Fibrosis

A disintegrin and metalloprotease 9 has been reported to play a key role in many diseases. However, the underlying effects and mechanisms of a disintegrin and metalloprotease 9 in alcoholic liver fibrosis have not been well understood. To investigate the role of a disintegrin and metalloprotease 9 and its regulatory molecular mechanism in alcoholic liver fibrosis, a single guide ribonucleic acid targeting the a disintegrin and metalloprotease 9 sequence of mice was adopted by clustered regularly interspaced short palindromic repeats/Cas9 technology to inhibit its expression in this study. Subsequently, hepatic stellate cell-T6 and male mice C57BL/6J were respectively divided into 4 groups and each group was treated differently. In vivo experiment, the normal group was fed with control Lieber-DeCarli TP4030C; the saline+alcohol group was injected with saline; the A disintegrin and metalloprotease 9-single guide ribonucleic acid+alcohol group was injected with effective single guide ribonucleic acid plasmids and the c-Jun N-terminal kinase inhibitor+alcohol group was injected with c-Jun N-terminal kinase inhibitor SP600125. All mice except for the normal group received Lieber-DeCarli TP4030A and carbon tetrachloride. Furthermore, examination of serum aspartate transaminase and alanine transaminase activities and the pathological analysis by hematoxylin-eosin staining, picric acid-sirius red staining, Hoechst 33 258 staining confirmed the pro-fibro genic effect of a disintegrin and metalloprotease 9. Compared with the saline+alcohol group, the levels and the expression of a disintegrin and metalloprotease 9, p-c-Jun, cytochrome P450 2E1, tumor necrosis factor alpha, alpha-smooth-muscle actin and Bcl-2-associated X in the A disintegrin and metalloprotease 9- single guide ribonucleic acid+alcohol group and the c-Jun N-terminal kinase inhibitor+alcohol group decreased significantly (p<0.05 or p<0.01), while the expression of heat shock protein 70, proliferating cell nuclear antigen and vascular endothelial growth factor increased significantly (p<0.05 or p<0.01). In addition, the disintegrin and metalloprotease 9-single guide ribonucleic acid+alcohol group and the c-Jun N-terminal kinase inhibitor+alcohol group in vitro experiment could inhibit the proliferation of hepatic stellate cells and promote apoptosis. Collectively, our results indicated that a disintegrin and metalloprotease 9 could promote alcoholic liver fibrosis in mice by activating the c-Jun N-terminal kinase signaling pathway, which is expected to make a breakthrough in the gene therapy of alcoholic liver fibrosis.

In vitro and In silico studies of interactions of cathinone with human recombinant cytochrome P450 CYP(1A2), CYP2A6, CYP2B6, CYP2C8, CYP2C19, CYP2E1, CYP2J2, and CYP3A5

Cathinone is the psychostimulatory major active ingredient of khat (Catha edulis Forsk) and are often co-abused with alcohols and polydrugs. With the increased consumption of khat and cathinones on a global scale, efforts should be channelled into understanding and minimising the excruciating effects of possible khat-drug interactions. This study aimed to determine the in vitro inhibitory effects of cathinone on CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C19, CYP2E1, CYP2J2 and CYP3A5 and the in silico identification of their type of interactions and residues involved. The activities of CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C19, CYP2E1, CYP2J2 and CYP3A5 were examined by fluorescence based assays using recombinant cDNA-expressed human CYPs in Vivid® P450 screening kits. Cathinone reversibly inhibited CYP1A2, CYP2A6 and CYP3A5 via competitive, uncompetitive and noncompetitive modes with inhibition constant (Ki) values of 57.12, 13.75 and 23.57 µM respectively. Cathinone showed negligible inhibitory effects on CYP2B6, CYP2C8, CYP2C19, CYP2E1 and CYP2J2. Cathinone showed negligible time dependent inhibition on all 8 CYPs. Docking studies was performed on cathinone with CYP1A2, CYP2A6 and CYP3A5 following their inhibition in vitro. Cathinone is bound to a few key amino acid residues in the active sites while π-π interactions are formed in aromatic clusters of CYP1A2 and CYP3A5. These findings offer valuable reference for the use of cathinones and khat when combined with therapeutic drugs that are metabolised by CYP enzymes especially patients on medications metabolised by CYP1A2, CYP2A6 and CYP3A5.

A combined metabolomics and molecular biology approach to reveal hepatic injury and underlying mechanisms after chronic l-lactate exposure in mice

This study aimed to explore whether chronic l-lactate exposure could affect the peripheral tissues of mice and to determine the underlying pathogenesis. Herein, male C57BL/6 mice were divided into control and l-lactate groups. After l-lactate treatment for eight weeks (1 g/kg), metabolic changes in liver, kidney, muscle, and serum samples were determined by 1H nuclear magnetic resonance (1H NMR)-based metabolomics. Additionally, organ function was evaluated by serum biochemical and histopathological examinations. Reactive oxygen species (ROS) levels were measured using dihydroethidium staining; levels of signals involved in lactate metabolism and ROS-related pathways were detected using western blotting or polymerase chain reaction. Apoptosis was detected by TUNEL-fluorescence staining. Metabolomic analysis revealed that l-lactate mice showed decreased levels of glutathione (GSH), taurine, ATP, and increased glucose content, compared to control mice. Furthermore, l-lactate mice presented significantly higher serum levels of alanine aminotransferase and aspartate aminotransferase and increased glycogen content in hepatic tissues, compared to control mice. l-lactate mice also had a greater number of apoptotic nuclei in the livers than controls. Moreover, l-lactate exposure reduced mRNA and protein levels of superoxide dismutase-2 and c-glutamylcysteine ligase, elevated levels of cytochrome P450 2E1 and NADPH oxidase-2, and increased the protein expressions of LDHB, Bax/Bcl-2, cleaved caspase-3, and sirtuin-1 in hepatic tissues. Together, these results indicate that chronic l-lactate exposure increases oxidative stress and apoptosis in hepatocytes via upregulation of Bax/Bcl-2 expression and the consequent mitochondrial cytochrome-C release and caspase-3 activation, which contributes to the pathogenesis of hepatic dysfunction.

Hepatoprotective Potential of Malabar Tamarind (Garcinia gummi-gutta) Fruit-Rind Extract in Acetaminophen-Induced Toxicity Through Modulation of CYP2E1 Gene Expression in Hepatic Cells of Wistar Rats

The present study was aimed to assess the hepatoprotective effect of ethanolic extract of Garcinia gummi-gutta (GGE) fruit rind on acetaminophen-induced toxicity in a rat model and to explore the possible mechanism of action by assessing hepatic histopathology, serum bio­chemical parameters and expression of CYP2E1 gene in hepatic cells. The group I and 11 animals served as normal control and hepatotoxic control group, respectively; while group III, IV and V were administrated with GGE fruit-rind extract @ 50, 100 & 200 mg kg-1 orally, respectively, for 10 days. The group VI (positive control) received drug silymarin @ 100 mg kg-1 day-1 orally for 10 days. Hepatotoxicity was induced by single oral administration of acetaminophen (2 g kg-1) on 8th day. The results revealed that the extract produced dose-dependent hepatoprotective effect against acetaminophen-induced hepatic damage by improving the serum bio­chemical profile towards the normal range. Histopathology of liver showed that pretreatment with fruit-rind extract attenuated the toxicant-induced hepatocellular necrosis and enhanced the regeneration of hepatic cells. The hepatic expression level of CYP2E1 gene was found down-regulated in a dose dependent manner in extract-treated group as compared to the normal control group. The study revealed remarkable hepatoprotective effect of GGE in acetaminophen-induced toxicity in rats through regeneration of hepatic cells and by decreasing the formation of reactive metabolite, NAPQ1 by modulation of CYP2E1 gene expression in liver.

РОЛЬ ИЗОФОРМ ЦИТОХРОМА Р450 ЭНДОПЛАЗМАТИЧЕСКОГО РЕТИКУЛУМА ГЕПАТОЦИТОВ В МЕТАБОЛИЗМЕ ЭТАНОЛА

Background. Three metabolic pathways that can function simultaneously are known to be involved in ethanol oxidation in the liver: alcohol dehydrogenase pathway, microsomal ethanol-oxidizing system, and catalase pathway. Though the cytochrome P450-dependent microsomal ethanol-oxidizing system plays an insignificant role in metabolism of small amounts of ethanol, it is induced in case of ethanol excess and becomes essential when ethanol is abused. The main components of this system are cytochrome P450 (CYP) isoforms of smooth endoplasmic reticulum. Objective. To characterize the role of the key isoforms of cytochrome P450 in ethanol oxidation. Material and methods. We carried out an analysis of modern literature data on the role of the main isoforms of cytochrome P450 in liver metabolism of ethanol. Results. Data on the primary role of cytochrome CYP2E1 in ethanol metabolism, as well as on the contribution of isoforms CYP1A2, CYP2B1/2, CYP2C, CYP3A4, CYP4B1 to ethanol oxidation are presented. Conclusions. Ethanol is metabolized by many CYPs of endoplasmic reticulum of hepatocytes. The importance of CYP in biotransformation processes in the liver necessitates the study of the role of individual CYP isoforms in ethanol metabolism for predicting changes in the pharmacokinetics of drugs and metabolism of endogenous compounds under the influence of ethanol.

Alcohol and Advanced Glycation End Product Synergistically Stimulate TNF-ɑ and TGF-β1 Production by Cytochrome P450 2E1 Expressing Macrophages

Alcohol and Advanced Glycation End Product Synergistically Stimulate TNF-ɑ and TGF-β1 Production by Cytochrome P450 2E1 Expressing Macrophages

The transcriptome of early GGT/KRT19-positive hepatocellular carcinoma reveals a downregulated gene expression profile associated with fatty acid metabolism

Hepatocellular carcinoma expressing hepatobiliary progenitor markers, is considered of poor prognosis. By using a hepatocarcinogenesis model, laser capture microdissection, and RNA-Sequencing analysis, we identified an expression profile in GGT/KRT19-positive experimental tumors; 438 differentially expressed genes were found in early and late nodules along with increased collagen deposition. Dysregulated genes were involved in Fatty Acid Metabolism, RXR function, and Hepatic Stellate Cells Activation. Downregulation of Slc27a5, Acsl1, and Cyp2e1, demonstrated that Retinoid X Receptor α (RXRα) function is compromised in GGT/KRT19-positive nodules. Since RXRα controls NRF2 pathway activation, we determined the expression of NRF2 targeted genes; Akr1b8, Akr7a3, Gstp1, Abcc3, Ptgr1, and Txnrd1 were upregulated, indicating NRF2 pathway activation. A comparative analysis in human HCC showed that SLC27A5, ACSL1, CYP2E1, and RXRα gene expression is mutually exclusive with KRT19 gene expression. Our results indicate that the downregulation of Slc27a5, Acsl1, Rxrα, and Cyp2e1 genes is an early event within GGT/KRT19-positive HCC.

Novel Sesquiterpene Glycoside from Loquat Leaf Alleviates Type 2 Diabetes Mellitus Combined with Nonalcoholic Fatty Liver Disease by Improving Insulin Resistance, Oxidative Stress, Inflammation, and Gut Microbiota Composition.

Nonalcoholic fatty liver disease (NAFLD) is strongly associated with type 2 diabetes mellitus (T2DM). Sesquiterpene glycosides from loquat leaf achieved beneficial effects on metabolic syndromes such as NAFLD and diabetes; however, their specific activity and underlying mechanism on T2DM-associated NAFLD have not yet been fully understood. In the present study, we found that sesquiterpene glycoside 3 (SG3), a novel sesquiterpene glycoside isolated from loquat leaf, was able to prevent insulin resistance (IR), oxidative stress, and inflammation. In db/db mice, SG3 administration (25 and 50 mg/kg/day) inhibited obesity, hyperglycemia, and the release of inflammatory cytokines. SG3 (5 and 10 μM) also significantly alleviated hepatic lipid accumulation, oxidative stress, and inflammatory response induced by high glucose combined with oleic acid in HepG2 cells. Western blotting analysis showed that these effects were related to repair the abnormal insulin signaling and inhibit the cytochrome P450 2E1 (CYP2E1) and NOD-like receptor family pyrin domain-containing 3 (NLRP3), both in vivo and in vitro. In addition, SG3 treatment could decrease the ratio of Firmicutes/Bacteroidetes and increase the relative abundance of Lachnospiraceae, Muribaculaceae, and Lactobacillaceae after a high-throughput pyrosequencing of 16S rRNA to observe the changes of related gut microbial composition in db/db mice. These findings proved that SG3 could protect against NAFLD in T2DM by improving IR, oxidative stress, inflammation through regulating insulin signaling and inhibiting CYP2E1/NLRP3 pathways, and remodeling the mouse gut microbiome. It is suggested that SG3 could be considered as a new functional additive for a healthy diet.

Relationships among N,N-dimethylformamide exposure, CYP2E1 and TM6SF2 genes, and non-alcoholic fatty liver disease

ObjectiveThis study aimed to examine the relationships among N, N-dimethylformamide (DMF) exposure, cytochrome P4502E1 (CYP2E1) single nucleotide polymorphisms (SNPs) (rs2031920, rs3813867, rs6413432), transmembrane 6 superfamily member 2 (TM6SF2) SNP rs58542926 and non-alcoholic fatty liver disease (NAFLD). MethodsBaseline data were collected from participants who were then followed for 5 years in a prospective cohort study. The cohort initially consisted of 802 workers and ultimately included 660 people, all of whom underwent annual occupational health examinations from 2010 to 2015. ResultsThe above-threshold group (≥7.3 mg/m³ adjusted relative risk (RR)= 3.620, 95%CI 2.072–6.325) was significantly more likely to develop NAFLD than the below-threshold group (<7.3 mg/m³). The TM6SF2 SNP rs58542926 CT (adjusted RR=3.921, 95% CI 2.329–6.600, P = 0.000) and CT+TT (adjusted RR=4.385, 95% CI 2.639–7.287, P = 0.000) genotypes were risk factors for NAFLD, as compared with the TM6SF2 rs58542926 CC genotype. Each dose group (below-threshold group and above-threshold group) interacting with the genotype of TM6SF2 SNP rs58542926 had an adjusted RR from 7.764 (95% CI 3.272–18.420, P = 0.000) to 24.022 (95% CI 8.971–64.328, P = 0.000). The T allele of rs58542926 in the TM6SF2 gene may be a risk factor for susceptibility to DMF-induced NAFLD. ConclusionPolymorphisms of TM6SF2 SNP rs58542926 may play an important role in susceptibility to NAFLD after exposure to DMF.

The Polymorphism and Expression of CYP2E1 Gene and Its Relation to Carcass and Meat Quality of Indonesian Lamb

The aim of this study was to conduct the genotyping of the single nucleotide polymorphism (SNP) in g.50657948 T>G of the CYP2E1 gene and its relation to the carcass and meat quality. A total of 200 Indonesian lambs consisted of 20 Javanese fat-tail sheep (JFTS), 37 Javanese thin-tail sheep (JTTS), 20 Garut sheep (GS), 21 Jonggol sheep (JS), 34 Garut composite sheep (GCS), 35 compass agrinac sheep (CAS), and 33 Barbados cross sheep (BCS) aged between 10-12 months old were used in this study. The polymorphism of the CYP2E1 gene was characterized using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with NlaIII enzyme restriction. The CYP2E1 gene found the three genotypes (GG, GT, and TT) and the allele frequencies were in Hardy Weinberg equilibrium. The CYP2E1 gene was significantly associated with meat quality, i.e., pH value and tenderness, as well as fatty acid composition (p<0.05), i.e., saturated fatty acid (SFA): caprylic acid (C8:0), arachidic acid (C20:0), heneicosylic acid (C21:0), behenic acid (C:22:0), and tricosylic acid (C23:0), monounsaturated fatty acid (MUFA): elaidic acid (C18:1n9t) and paullinic acid (C20:1); polyunsaturated fatty acid (PUFA): linoleic acid (C18:2n6c) and y-linolenic acid (C20:3n6). A gene expression analysis indicated that the GG genotype had the highest expression level. It could be concluded that the SNP g.50657948 T>G of the CYP2E1 gene could be used for candidate marker-assisted selection to improve the carcass and meat quality of Indonesian lamb.

Genetic Polymorphism of Drug Metabolic Gene CYPs, VKORC1, NAT2, DPYD and CHST3 of Five Ethnic Minorities in Heilongjiang Province, Northeast China.

IntroductionGenetic variability in genes encoding drug-metabolizing enzymes may contribute to the heterogeneity of drug responses in different populations. Extensive research in pharmacogenomics in major populations around the world provides us with a great deal of information about drug-related genetic polymorphisms.ObjectiveThe purpose of this study was to detect the genetic variation of drug-metabolism-related genes in the five ethnic minorities Daur, Hezhen, Ewenki, Mongolian and Manchu in China, and to analyze the distribution differences among ethnic groups.MethodsWe genotyped 32 SNPs of drug metabolism genes in 882 healthy Chinese volunteers from five ethnic groups. The genotype frequency and allele frequency of the five ethnic groups were calculated, and the different variants among the five ethnic groups were compared by chi-square test. Genetic parameters were analyzed using Popgene software. The genetic structure of five ethnic minorities was analyzed by principal component analysis, and compared with 26 populations.ResultsWe found that SNPs of genes related to drug metabolism existed diversity in different populations. Among them, rs8192766 and rs9419082 in CYP2E1 showed statistical differences between Daur and Manchu, and NAT2 rs1801280 showed statistical differences between Hezhen and Mongolian. In addition, the five populations we studied had the smallest differences with EAS populations. There was haplotype diversity in CHST3, VKORC1, CYP1A2 and CYP2E1 genes in the five ethnic minorities, and these haplotype polymorphisms were related to the use of corresponding drug doses. Cluster analysis shows that the five ethnic minorities in Heilongjiang Province are clustered together with the EAS populations.ConclusionThese results suggest that understanding the diversity of drug-related genetic markers is critical for individualized drug gene therapy programs in ethnic minorities in China as well as in populations highly mixed with these ethnic groups.