CYP2E1 in Alcoholic and Non-Alcoholic Liver Injury. Roles of ROS,Reactive Intermediates and Lipid Overload

CYP2E1 is one of the fifty-seven cytochrome P450 genes in the human genome and is highly conserved. CYP2E1 is a unique P450 enzyme because its heme iron is constitutively in the high spin state, allowing direct reduction of, e.g., dioxygen, causing the formation of a variety of reactive oxygen species and reduction of xenobiotics to toxic products. The CYP2E1 enzyme has been the focus of scientific interest due to (i) its important endogenous function in liver homeostasis, (ii) its ability to activate procarcinogens and to convert certain drugs, e.g., paracetamol and anesthetics, to cytotoxic end products, (iii) its unique ability to effectively reduce dioxygen to radical species causing liver injury, (iv) its capability to reduce compounds, often generating radical intermediates of direct toxic or indirect immunotoxic properties and (v) its contribution to the development of alcoholic liver disease, steatosis and NASH. In this overview, we present the discovery of the enzyme and studies in humans, 3D liver systems and genetically modified mice to disclose its function and clinical relevance. Induction of the CYP2E1 enzyme either by alcohol or high-fat diet leads to increased severity of liver pathology and likelihood to develop ALD and NASH, with subsequent influence on the occurrence of hepatocellular cancer. Thus, fat-dependent induction of the enzyme might provide a link between steatosis and fibrosis in the liver. We conclude that CYP2E1 has many important physiological functions and is a key enzyme for hepatic carcinogenesis, drug toxicity and liver disease.     

An Advanced Electron Spin Resonance (ESR) Spin-Trapping and LC/(ESR)/MS Technique for the Study of Lipid Peroxidation

There are two types of nutritionally important polyunsaturated fatty acids (PUFAs), namely ω-6s and ω-3s. PUFAs and their metabolites generated from lipid peroxidation via cyclooxygenase (COX) and lipoxygenase (LOX) are believed to be involved in a variety of physiological and pathological processes in the human body. Both COX- and LOX-catalyzed PUFA peroxidation are complex events that generate a series of radicals, which may then bind proteins, target DNA/RNA, and lead to a number of biological changes. However, due to the lack of an appropriate method, it was not possible until recently to identify the short-lived PUFA-derived radicals in COX-/LOX-catalyzed peroxidation. Failure to characterize free radicals during peroxidation has greatly restricted our knowledge about COX/LOX biology in human health. Here we review the development and refinement of combined ESR spin trapping and LC/ESR/MS to characterize PUFA-derived radicals formed from in vitro (cell-free) peroxidation. We also present the most recent approach for studying peroxidation in cells which allows us to directly assess the potential bioactivity of PUFA-derived free radicals. This advanced technique has resulted in a major breakthrough in radical structural characterization, as well as assessment of free radical-associated cell growth response, thereby greatly improving our knowledge of PUFAs, COX-/LOX-catalyzed lipid peroxidation, and their related biological consequences.     

Lipidomic Profiling Reveals the Effect of Egg Components on Nonalcoholic Steatosis in HepG2 Cells and Its Involved Mechanisms

Non‐alcoholic fatty liver disease (NAFLD) is a major cause of cardiovascular disease. The relationship between egg consumption and NAFLD is still controversial for its high cholesterol content. In this study, the effects of different egg components (egg white (EW), egg yolk (EY), and whole egg (WE)) on NAFLD are examined using oleic acid (OA)‐induced HepG2 cells with UPLC‐ESI‐MS/MS approach. The results show EY could affect the lipid profile effectively by increasing phosphocholine (PC), phosphatidylglycerol (PG), and carnitine (CAR). Orthogonal projections to latent structures?discriminate analysis (OPLS‐DA) combine with S‐plot analysis select 10, 82, and 20 potential biomarkers in EW, EY, and WE group, respectively. Up‐regulated TG, DG, and down‐regulated lysophosphatidylcholines (lysoPC), lysophosphatidylethanolamine (lysoPE) biomarkers are found in EY group, while down‐regulated TG and FFA are found in EW and WE group. Glycerolipid and choline metabolism are the most involved pathways affected by EY and WE. In addition, the mechanism is associated with the expression of Pla2g15, Pnpla6‐1, Gad1, and involved lipogenic genes ABC1 and PPARα. This study suggests that WE treatment can ameliorate OA‐induced hepatic steatosis by inhibiting TG accumulation, while EY seems slightly accelerate hepatic steatosis. Furthermore, the effects are closely associated with its effects on glycerolipid metabolism. Practical applications: Nonalcoholic fatty liver disease (NAFLD) is a worldwide disease, while the associations between egg consumption and NAFLD are still poorly understood. This study investigates the effects of egg components on NAFLD in oleic acid (OA)‐induced HepG2 cells based on a targeted lipidomics approach. The results indicate that WE (whole egg) treatment could ameliorate OA‐induced hepatic steatosis by inhibiting TG and FFA accumulation, which is closely associated with glycerolipid metabolism. The results provide knowledge and understanding of the effects of egg on NAFLD and involved mechanism, and further provided nutritional guidelines for egg consumption.     

Ultra‐Performance Liquid Chromatography‐Mass Spectrometry Analysis of Free and Esterified Oxygenated Derivatives from Docosahexaenoic Acid in Rat Brain

Docosahexaenoic acid (DHA), a prominent long‐chain fatty acid of the omega‐3 family, is present at high amount in brain tissues, especially in membrane phospholipids. This polyunsaturated fatty acid is the precursor of various oxygenated lipid mediators involved in diverse physiological and pathophysiological processes. Characterization of DHA‐oxygenated metabolites is therefore crucial for better understanding the biological roles of DHA. In this study, we identified and measured, by ultrahigh‐performance liquid chromatography coupled with tandem mass spectrometry, a number of oxygenated products derived from DHA in exsanguinated and nonexsanguinated brains. These metabolites were found both in free form and esterified in phospholipids. Interestingly, both (R)‐ and (S)‐monohydroxylated fatty acid stereoisomers were observed free and esterified in phospholipids. Monohydroxylated metabolites were the main derivatives; however, measurable amounts of dihydroxylated products such as protectin DX were detected. Moreover, exsanguination allowed discriminating brain oxygenated metabolites from those generated in blood. These results obtained in healthy rats allowed an overview on the brain oxygenated metabolism of DHA, which deserves further research in pathophysiological conditions, especially in neurodegenerative diseases.     

In Vitro Antioxidant Properties of Berry Leaves and Their Inhibitory Effect on Lipid Peroxidation of Thigh Meat from Broiler Chickens

The present study is designed to evaluate the antioxidant compounds from bilberry, cranberry, and raspberry leaves in connection with their radical scavenging activities, and their potential in inhibiting the lipid peroxidation of thigh meat from broiler chickens. For this purpose, plant extracts are analyzed regarding the content in bioactive compounds, antioxidant properties analysis including 1,1‐diphenyl‐2‐picrylhydrazyl, superoxide, hydrogen peroxide, and hydroxyl radical‐scavenging activities. Lipid peroxidation is induced by an Fe⁺³/ascorbic acid system and the inhibitory effects of plants extracts are assessed using thiobarbituric reactive substances (TBARS) as biomarkers for quantifying lipid oxidation in meat. The overview of the analyzed antioxidant compounds shows that cranberry leaves present the highest concentrations of vitamin E, β‐carotene, lutein and zeaxanthin, and polyphenols and a total antioxidant capacity with 52% higher than bilberry and 360% than raspberry. Bilberry leaves extract shows the strongest effect (p  < 0.0001) in inhibiting meat lipid peroxidation. Practical Applications: The results demonstrate the positive effect of the selected berry leaves in the oxidative stability of broiler chicken meat under in vitro conditions, and may be suggested as an alternative source of natural antioxidants for animal nutrition, with benefits in inhibiting and delaying oxidation, and also enhancing the nutritional value of meat.     

Fatty acids,sterols, and tocopherols composition in seed oil extracts from four moroccan ecotypes of Ceratonia siliqua L.

Morphological characterization was investigated by agro-morphological criteria related to carob seed size in four different moroccan regions collected in 2018 and 2019. There was no significant difference (p ≤ 0.05) on the seeds lengths and widths. However, a significant difference between seeds thickness and total seeds weight per pod (p ≤ 0.05) were observed between these four populations. The fatty acid, sterol, tocopherol, hydrocarbon, and the unoxygenated composition of carob seed extracts (Ceratonia siliqua L.) were studied. The mean fat yield of the seeds obtained is 1.53%–2.17%, 2.14%–2.15%, 1.61%–1.62%, 1.71%–1.75% for, respectively, the P1 (Meknes), P2 (Fez), P3 (Khemisset), and P4 (Marrakech) in 2018 and 2019. The seed oil was extracted with hexane and the analysis of the fatty fraction was performed by gas chromatography–mass spectrometry (GC–MS). Results show that the major fatty acids for 2018 and 2019 are linoleic acid (61.48%–61.52%, 52.12%–52.14%, 57.76%–58.15%, 61.33%–61.52%), palmitic acid (15.78%–15.81%, 16.44%–16.45%, 19.11%–18.37%, 20.24%–20.32%), oleic acid (11.03%–11.04%, 8.72%–8.82%, 8.51%–8.61%, 8.41%–8.53%), stearic acid (4.35%–3.14%, 5.40%–5.43%, 3.12%–3.13%, 0.96%–1.56%), and cerotic acid (0.62%–0.53%, 4.51%–4.52%, 4.03%–4.06%, 3.84%–3.87%). The unsaturated fatty acids (69.39% in 2018 and 69.68% in 2019) are the most dominant in the four seed extracts compared to the saturated fatty acids. In addition, the oil carob seeds analysis revealed the presence of γ-tocopherol, α-tocopherol and four sterols that included campesterol, stigmasterol, and β-sitosterol. Moreover, the determination of hydrocarbon and un-oxygenated compounds confirmed the existence of major compounds such as heptadecane, 2-methyltriacontane, 1-iodo hexadecane and 1-iodo octadecane. The hierarchical analysis based on the morphological and chromatographic characterization of the seeds allowed the identification of three groups. Consequently, the first group consisted of populations from Marrakesh (P4) and Khemisset (P3), the second group consisted of the P1 from Meknes, and the P2 from Fez constituted the third group.     

Extraction of phenolic compounds from sour cherry pomace with supercritical carbon dioxide: Impact of process parameters on the composition and antioxidant properties of extracts

Sour cherry (Prunus cerasus) is rich in biologically active phenolic compounds. These compounds are concentrated in fruit skin and most of them remain in the leftovers during the production of juice. Supercritical carbon dioxide extraction was used to separate phenolic compounds from sour cherry pomace. The effects of temperature, pressure and the addition of ethanol on anthocyanin and the total phenolic content and radical scavenging activity of the extracts were investigated. The best results were acquired for 35°C, 10 MPa and 80% ethanol addition. A strong correlation was found between the phenolic content and other features of the extracts.     

Plant photosensitizers: A survey of their occurrence in arid and semiarid plants from North America

Various plants native to arid and semiarid habitats throughout the southwestern United States, Baja California, and northern Mexico were bioassayed for phototoxic natural products. Approximately 115 species representing 57 genera and eight plant families were assayed for phototoxic activity by standard antimicrobial techniques usingEscherichia coli andSaccharomyces cerevisiae. Phototoxic constituents were extracted from numerous members in the Asteraceae (Compositae) and occurred with highest frequency among species of the subtribe Pectidinae (tribe Heliantheae). Extracts ofPectis, the largest genus in the Pectidinae, had substantial light-activated biocidal action despite the paucity of acetylenic thiophenes, the phototoxins characteristic of most other genera in the subtribe. Leaf resin from the creosote bush [Larrea tridentata (Sesse & Mol. ex DC.) Coville; Zygophyllaceae], a dominant desert shrub, possessed potent antimicrobial activity in the absence of light; however, the toxicity of this extract was slightly enhanced in the presence of UVA irradiation. Phototoxic antimicrobials were not detected in extracts of selected species from the Asclepiadaceae, Chenopodiaceae, Hydrophyllaceae, Lamiaceae, Polygonaceae, or Solanaceae.