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1.
Arthritis Res Ther ; 25(1): 5, 2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36609408

RESUMO

BACKGROUND: Behçet's disease (BD) is a systemic inflammatory disease that involves various organs. The clinical manifestation-based diagnosis of BD is a time-consuming process, which makes it difficult to distinguish from patients with similar symptoms. Moreover, an authentic biomarker has not been developed for accurate diagnosis yet. Our current study investigated the unique metabolic signatures of BD and explored biomarkers for precise diagnosis based on an untargeted metabolomic approach. METHODS: Integrative metabolomic and lipidomic profiling was performed on plasma samples of BD patients (n = 40), healthy controls (HCs, n = 18), and disease controls (DCs, n = 17) using GC-TOF MS and LC-Orbitrap MS. Additionally, the lipid profiles of 66 peripheral blood mononuclear cells (PBMCs) were analyzed from 29 BD patients, 18 HCs, and 19 DCs. RESULTS: Plasma metabolic dysfunction in BD was determined in carbohydrate, hydroxy fatty acid, and polyunsaturated fatty acid metabolisms. A plasma biomarker panel with 13 compounds was constructed, which simultaneously distinguished BD from HC and DC (AUCs ranged from 0.810 to 0.966). Dysregulated PBMC metabolome was signatured by a significant elevation in lysophosphatidylcholines (LPCs) and ether-linked lysophosphatidylethanolamines (EtherLPEs). Ten PBMC-derived lipid composites showed good discrimination power (AUCs ranged from 0.900 to 0.973). Correlation analysis revealed a potential association between disease activity and the metabolites of plasma and PBMC, including sphingosine-1 phosphate and EtherLPE 18:2. CONCLUSIONS: We identified metabolic biomarkers from plasma PBMC, which selectively discriminated BD from healthy control and patients with similar symptoms (recurrent mouth ulcers with/without genital ulcers). The strong correlation was determined between the BD activity and the lipid molecules. These findings may lead to the development for diagnostic and prognostic biomarkers based on a better understanding of the BD pathomechanism.


Assuntos
Síndrome de Behçet , Humanos , Síndrome de Behçet/metabolismo , Leucócitos Mononucleares/metabolismo , Metabolômica , Biomarcadores , Lipídeos , Estudos de Casos e Controles
2.
Nat Commun ; 14(1): 96, 2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36609407

RESUMO

Gated entry of lipophilic ligands into the enclosed hydrophobic pocket in stand-alone Sec14 domain proteins often links lipid metabolism to membrane trafficking. Similar domains occur in multidomain mammalian proteins that activate small GTPases and regulate actin dynamics. The neuronal RhoGEF Kalirin, a central regulator of cytoskeletal dynamics, contains a Sec14 domain (KalbSec14) followed by multiple spectrin-like repeats and catalytic domains. Previous studies demonstrated that Kalirin lacking its Sec14 domain fails to maintain cell morphology or dendritic spine length, yet whether and how KalbSec14 interacts with lipids remain unknown. Here, we report the structural and biochemical characterization of KalbSec14. KalbSec14 adopts a closed conformation, sealing off the canonical ligand entry site, and instead employs a surface groove to bind a limited set of lysophospholipids. The low-affinity interactions of KalbSec14 with lysolipids are expected to serve as a general model for the regulation of Rho signaling by other Sec14-containing Rho activators.


Assuntos
Actinas , Citoesqueleto , Animais , Fatores de Troca de Nucleotídeo Guanina Rho/genética , Lipídeos , Mamíferos
3.
Int J Mol Sci ; 24(1)2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36614187

RESUMO

Ladderane lipids (found in the membranes of anaerobic ammonium-oxidizing [anammox] bacteria) have unique ladder-like hydrophobic groups, and their highly strained exotic structure has attracted the attention of scientists. Although enzymes encoded in type II fatty acid biosynthesis (FASII) gene clusters in anammox bacteria, such as S-adenosyl-l-methionine (SAM)-dependent enzymes, have been proposed to construct a ladder-like structure using a substrate connected to acyl carrier protein from anammox bacteria (AmxACP), no experimental evidence to support this hypothesis was reported to date. Here, we report the crystal structure of a SAM-dependent methyltransferase from anammox bacteria (AmxMT1) that has a substrate and active site pocket between a class I SAM methyltransferase-like core domain and an additional α-helix inserted into the core domain. Structural comparisons with homologous SAM-dependent C-methyltransferases in polyketide synthase, AmxACP pull-down assays, AmxACP/AmxMT1 complex structure predictions by AlphaFold, and a substrate docking simulation suggested that a small compound connected to AmxACP could be inserted into the pocket of AmxMT1, and then the enzyme transfers a methyl group from SAM to the substrate to produce branched lipids. Although the enzymes responsible for constructing the ladder-like structure remain unknown, our study, for the first time, supports the hypothesis that biosynthetic intermediates connected to AmxACP are processed by SAM-dependent enzymes, which are not typically involved in the FASII system, to produce the ladder-like structure of ladderane lipids in anammox bacteria.


Assuntos
Metionina , S-Adenosilmetionina , S-Adenosilmetionina/metabolismo , Metionina/metabolismo , Proteína de Transporte de Acila/metabolismo , Metiltransferases/metabolismo , Oxidação Anaeróbia da Amônia , Bactérias/metabolismo , Racemetionina/metabolismo , Lipídeos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo
4.
Int J Mol Sci ; 24(1)2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36614230

RESUMO

Nanoparticles are heterologous small composites that are usually between 1 and 100 nanometers in size. They are applied in many areas of medicine with one of them being drug delivery. Nanoparticles have a number of advantages as drug carriers which include reduced toxic effects, increased bioavailability, and their ability to be modified for specific tissues or cells. Due to the exciting development of nanotechnology concomitant with advances in biotechnology and medicine, the number of clinical trials devoted to nanoparticles for drug delivery is growing rapidly. Some nanoparticles, lipid-based types, in particular, played a crucial role in the developing and manufacturing of the two COVID-19 vaccines-Pfizer and Moderna-that are now being widely used. In this analysis, we provide a quantitative survey of clinical trials using nanoparticles during the period from 2002 to 2021 as well as the recent FDA-approved drugs (since 2016). A total of 486 clinical trials were identified using the clinicaltrials.gov database. The prevailing types of nanoparticles were liposomes (44%) and protein-based formulations (26%) during this period. The most commonly investigated content of the nanoparticles were paclitaxel (23%), metals (11%), doxorubicin (9%), bupivacaine and various vaccines (both were 8%). Among the FDA-approved nanoparticle drugs, polymeric (29%), liposomal (22%) and lipid-based (21%) drugs were the most common. In this analysis, we also discuss the differential development of the diverse groups of nanoparticles and their content, as well as the underlying factors behind the trends.


Assuntos
COVID-19 , Nanopartículas , Humanos , Vacinas contra COVID-19/uso terapêutico , COVID-19/prevenção & controle , Lipossomos , Lipídeos
5.
PLoS One ; 18(1): e0279977, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36652431

RESUMO

PURPOSE: To prospectively assess the effect of a single and regular application of either a cationic nanoemulsion of mineral oil (CN) or perfluorohexyloctane (F6H8) on the lipid layer of the tear film and higher order aberrations (HOA) in patients with Dry Eye Disease (DED). METHODS: Fifty-seven patients with a lipid layer thickness (LLT) ≤ 75 interferometric colour units (ICU) were included in the study. In group A (20 patients) the effect of a single drop of F6H8 or CN on HOA and LLT was assessed immediately after application and up to two hours later. For long term effects (Group B) 37 patients applied CN or F6H8 five times a day for 12 weeks. Measurement of LLT, HOA, non-invasive-tear-break-up-time (NIBUT) and meibography were assessed prior to as well as at 4 weeks and 12 weeks after initiation of treatment. Our study is registered in the "German Clinical Trials Register" under the trial number: DRKS00028696. RESULTS: CN led to an increase of the LLT from 46.8 ± 16.9 ICU to 76.3 ± 23.5 ICU (p = 0.021) and to an increase of HOA from 0.43 ± 0.06 µm to 0.48 ± 0.08 µm immediately after application (p = 0.027). There was no correlation between the increase of LLT and HOA (r = -0.04; p = 0.90). In group B an increase of LLT was observed in the F6H8 group from 45.8 ± 8.8 ICU at baseline to 66.7 ± 19.5 ICU at 12 weeks (p = 0.002). No changes of HOA were measured throughout the observation period in group B. After 12 weeks CN increased NIBUT from 9.9 ± 5.3 seconds to 15.5 ± 5.6 seconds (p = 0.04). F6H8 increased NIBUT from 12.4 ± 5.9 seconds to 16.9 ± 4.7 seconds (p = 0.02) after 12 weeks. CONCLUSION: CN leads to a short-term increase in LLT and HOA, but only immediately after application. In contrast F6H8 does lead to an increase of LLT after regular long-term use but has no effect on HOA. The regular application of lipid-based products does not seem to decrease the quality of vision as measured in HOA. Instead, CN and F6H8, both are able to stabilize the tear film after regular application.


Assuntos
Síndromes do Olho Seco , Fluorcarbonetos , Lacerações , Humanos , Síndromes do Olho Seco/tratamento farmacológico , Fluorcarbonetos/uso terapêutico , Lipídeos/uso terapêutico , Óleo Mineral , Lágrimas
6.
Sci Adv ; 9(2): eadc8825, 2023 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-36638181

RESUMO

Metastatic disease is a major cause of death for patients with melanoma. Melanoma cells can become metastatic not only due to cell-intrinsic plasticity but also due to cancer-induced protumorigenic remodeling of the immune microenvironment. Here, we report that innate immune surveillance by natural killer (NK) cells is bypassed by human melanoma cells expressing the stem cell marker NGFR. Using in vitro and in vivo cytotoxic assays, we show that NGFR protects melanoma cells from NK cell-mediated killing and, furthermore, boosts metastasis formation in a mouse model with adoptively transferred human NK cells. Mechanistically, NGFR leads to down-regulation of NK cell activating ligands and simultaneous up-regulation of the fatty acid stearoyl-coenzyme A desaturase (SCD) in melanoma cells. Notably, pharmacological and small interfering RNA-mediated inhibition of SCD reverted NGFR-induced NK cell evasion in vitro and in vivo. Hence, NGFR orchestrates immune control antagonizing pathways to protect melanoma cells from NK cell clearance, which ultimately favors metastatic disease.


Assuntos
Antineoplásicos , Melanoma , Camundongos , Animais , Humanos , Linhagem Celular Tumoral , Melanoma/patologia , Células Matadoras Naturais , Lipídeos , Microambiente Tumoral , Proteínas do Tecido Nervoso/metabolismo , Receptores de Fator de Crescimento Neural/metabolismo
7.
Invest Ophthalmol Vis Sci ; 64(1): 13, 2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36656568

RESUMO

Purpose: A majority of in vitro models were incapable of reproducing the evaporation resistance of tear film lipid layer (TFLL) in vivo. The purpose of this research is to develop a novel in vitro model to study the effect of TFLL on water evaporation. Methods: A ventilated, closed-chamber, droplet evaporimeter with a constant surface area has been invented to study the evaporation resistance of TFLL. This evaporimeter ensures a rigorous control of environmental conditions, including the temperature, relative humidity, airflow rate, surface area, and surface pressure, thus allowing for reproducible water evaporation measurements over a time period of only 5 minutes. The volumetric evaporation rate of this droplet evaporimeter is less than 2.7 µL/min, comparable to the basal tear production of healthy adults. Together with direct film imaging using atomic force microscopy (AFM), we have studied the effect of a model TFLL on water evaporation, as a function of the lipid composition and surface pressure. Results: A model TFLL composed of 40% wax esters, 40% cholesteryl esters, and 20% polar lipids was capable of reducing the water evaporation rate by 11% at surface pressure 47 mN/m. AFM revealed that the model TFLL at high surface pressures consists of discrete droplets/aggregates of the nonpolar lipids residing atop a polar lipid monolayer with phase separation. Conclusions: The TFLL may resist water evaporation with a combined mechanism by increasing film compactness of the polar lipid film at the air-water surface, and, to a lesser extent, by increasing film thickness of the nonpolar lipid film.


Assuntos
Lipídeos , Água , Humanos , Adulto , Ésteres do Colesterol , Lágrimas
8.
Artigo em Inglês | MEDLINE | ID: mdl-36673736

RESUMO

Hypertension and dyslipidemia are major risk factors for cardiovascular disease. Studies on the association between abnormal levels of lipids and hypertension have yielded inconsistent results. The aim of this study was to examine the prevalence of hypertension and dyslipidemia and its risk factors in young Polish adults. Furthermore, the association between plasma lipid levels and the risk of hypertension was determined. A cross-sectional study was conducted among 115 volunteer participants. Blood pressure was measured using an automated oscillometric sphygmomanometer. Blood lipids were analyzed from a fasting blood sample received by finger prick. Body fat percentage was assessed using a bioelectrical impedance analysis device. Socioeconomic and lifestyle factors (age, date of birth, place of residence, screen time, and tobacco use) were self-reported by the participant. The prevalence of hypertension was higher in men than in women (61.5 vs. 21.3%). The prevalence of elevated TC, TG, high LDL, and low HDL was 22.6%, 7.8%, 38.3%, and 13.9%, respectively. Spending more than 2 h daily in front of a computer was identified as a significant predictor of hypertension and elevated TG levels (p < 0.05). A high number of cigarettes smoked daily was a significant risk factor for hypertension (p = 0.047). Hypertension contributed to a higher risk of abnormal values of TC (OR = 5.89), LDL (OR = 5.38), and TG (OR = 9.75). Participants with hypertension were more likely than normotensive subjects to have elevated levels of TC, LDL, and TG. The prevalence of hypertension was significantly higher in young men than in women. BMI was associated with the prevalence of hypertension and elevated TC levels. Spending more than 2 h per day in front of a computer contributed to the prevalence of hypertension and elevated TG levels. Participants with hypertension smoked a higher number of cigarettes daily compared to those with normotension.


Assuntos
Dislipidemias , Hipertensão , Masculino , Humanos , Feminino , Adulto Jovem , Polônia/epidemiologia , Estudos Transversais , Hipertensão/complicações , Fatores de Risco , Lipídeos , Dislipidemias/epidemiologia , Dislipidemias/complicações , Triglicerídeos , Prevalência
9.
Molecules ; 28(2)2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36677770

RESUMO

Propolis is a resinous compound made by bees with well-known biological activity. However, comparisons between encapsulated and non-encapsulated propolis are lacking. Therefore, the antibacterial activity, effect on the phase transition of lipids, and inhibition of UV-induced lipid oxidation of the two forms of propolis were compared. The results showed that non-encapsulated propolis produces quicker effects, thus being better suited when more immediate effects are required (e.g., antibacterial activity). In order to gain an in-depth introspective on these effects, we further studied the synergistic effect of propolis compounds on the integrity of lipid membranes. The knowledge of component synergism is important for the understanding of effective propolis pathways and for the perspective of modes of action of synergism between different polyphenols in various extracts. Thus, five representative molecules, all previously isolated from propolis (chrysin, quercetin, trans-ferulic acid, caffeic acid, (-)-epigallocatechin-3-gallate) were mixed, and their synergistic effects on lipid bilayers were investigated, mainly using DSC. The results showed that some compounds (quercetin, chrysin) exhibit synergism, whereas others (caffeic acid, t-ferulic acid) do not show any such effects. The results also showed that the synergistic effects of mixtures composed from several different compounds are extremely complex to study, and that their prediction requires further modeling approaches.


Assuntos
Própole , Própole/farmacologia , Quercetina/farmacologia , Flavonoides/farmacologia , Bactérias , Antibacterianos/farmacologia , Lipídeos
10.
Nanotheranostics ; 7(1): 117-127, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36593793

RESUMO

Background: Irbesartan (IR) is used in the treatment of hypertension, heart failure, and nephropathy in Type II diabetes. IR bioavailability is limited by poor solubility and presystemic metabolism. In our previous investigations, cyclodextrin (HPßCD) complexed solid lipid nanoparticles (SLNs) of IR were prepared, optimized, and characterized. The current study aimed to confirm the reproducibility of the previous methodology and to evaluate the pharmacokinetic (PK) and pharmacodynamic (PD) performance of the selected lead formulations in an experimental animal model. Methods: SLNs were prepared by hot homogenization followed by probe sonication with IR/HPßCD inclusion complex loaded into a solid lipid (Dynasan 112). SLNs were evaluated for physical characteristics, drug content, entrapment efficiency, in vitro release profile, and surface morphology. The pharmacokinetic and pharmacodynamic behavior of the SLNs were evaluated in Wistar rats. Results: Photon correlation spectroscopy, drug content, entrapment efficiency, and dissolution studies results were reproducible and consistent with our earlier investigation. PK studies showed 2.1-, 6.6-, and 9.9-fold improvement in the relative oral bioavailability of the drug from IR-HPßCD, IR-SLN, and IR-HPßCD-SLN formulations, respectively compared to IR suspension. However, IR-HPßCD-SLNs showed 1.5- and 4.7-fold improvement in the relative oral bioavailability of the drug compared to IR-SLN and IR-HPßCD formulations, respectively. PD studies in hypertensive Wistar rats showed a good control over systolic blood pressure for 48 h for SLN formulations compared to 2 h for IR suspension. However, the IR-HPßCD inclusion complex exhibited immediate antihypertensive activity (0.5 h) with a period of systolic blood pressure control similar to IR suspension. Conclusions: The current approach exhibited improved oral bioavailability along with improved and prolonged pharmacodynamic effect.


Assuntos
Ciclodextrinas , Diabetes Mellitus Tipo 2 , Ratos , Animais , Ratos Wistar , Disponibilidade Biológica , Irbesartana/farmacologia , Lipídeos/química , Portadores de Fármacos/química , 2-Hidroxipropil-beta-Ciclodextrina , Ciclodextrinas/farmacologia , Reprodutibilidade dos Testes
11.
Nanotheranostics ; 7(1): 91-101, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36593795

RESUMO

Stress can be defined by two parameters, first the psychological sensing of pressure and second is the body's response. However, the exposure time to stress depicts the biological response produced against it. The effect of acute and chronic restraint stress on anxiety and the production of systemic metabolites were investigated in male Sprague-Dawley (SD) rats. Behavioural test was performed on elevated plus maze (EPM) in conjunction with the statistical analysis that exhibited the habituation during long term exposure to stress when compared with the short-term stress. These behaviour-based changes resulted in interpolated concentration of some serum metabolites like carbohydrates, amino acids and lipids as analysed by NMR. Metabolic analysis along with the multivariate analysis demonstrated that the expression of concentration of metabolites including glutamate, proline, succinate, citrate, and tyrosine is higher in the acute stress than the chronic stress, while glucose and lipids i.e., LDL and VLDL changed in the opposite trends. Thus, the aforesaid study provides an analytical strategy for the characterization of perturbed metabolites induced due to the behavioural modifications in an organism. It may further aid in developing potential therapeutic markers at the metabolic levels which may broaden the treatment options for stress and anxiety related disorders.


Assuntos
Imageamento por Ressonância Magnética , Restrição Física , Ratos , Animais , Masculino , Ratos Sprague-Dawley , Espectroscopia de Ressonância Magnética , Lipídeos
12.
Methods Mol Biol ; 2625: 291-298, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36653651

RESUMO

Lipids are among the major constituents of cells and play many important cellular functions. Lipid levels in the trabecular meshwork (TM) aqueous humor outflow pathway play an important role in the maintenance of aqueous humor drainage and intraocular pressure (IOP) homeostasis. Therefore, it is important to characterize the changes in the lipid contents in the aqueous humor outflow pathway tissues to better understand their functional significance in the maintenance of IOP. The multiple reaction monitoring (MRM)-based profiling aids in the analysis of the metabolome as a collection of functional groups and is utilized as an exploratory metabolomics and lipidomics approach. The MRM-based profiling utilizes tandem mass spectrometry experiments carried out on a commercial triple quadrupole mass spectrometer with three aligned quadrupole mass filters (Q1, Q2, and Q3). This screening methodology can be utilized for targeted lipidomics screening. This chapter focuses on the methodology for isolation and culturing of the TM cells, lipid extraction, and the MRM-based lipidomics approach with data analysis.


Assuntos
Lipidômica , Malha Trabecular , Humanos , Malha Trabecular/metabolismo , Humor Aquoso/metabolismo , Pressão Intraocular , Lipídeos
13.
Eur J Med Chem ; 247: 115053, 2023 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-36587419

RESUMO

Herein 2-cyanoethoxy-N,N,N',N'-tetraisopropyl-phosphorodiamidite(10, PIII, 3.5 eq.) could synergistically react with 3',5'-dihydroxyl groups in a dinucleotide(PV) at the cyclization step for the synthesis of cyclic dinucleotides (CDNs) (c-di-GMP, cGAMP etc.) and their phosphorothioated analogues. A dynamic PIII-PV coordination mechanism has been proposed for the cyclization procedure which is confirmed by the variant 31P NMR data and molecular simulation. Among the mono-phosphorothioated CDNs, two stereoisomers showed different capacity for STING activation and the reason was predicted by molecular modeling. While compound 12b1 showed most potent ability to elicit cytokines (IFNß, IL-6, Cxcl9 and Cxcl10) induction compared to another stereoisomer. Also, 12b1 significantly inhibited the tumor growth in the EO771 model with both 0.1 µg (i.t.) and 2 µg (i.v.) administration through the aid of a Mix delivery system developed by our group, and achieved a 31% long-term survival rate of tumor-bearing mice. 12b1/Mix significantly improved the percentage of CD8+ or CD4+ effector memory T (Tem, CD44highCD62Llow) cells and CD8+ central memory T (Tcm, CD44highCD62Lhigh) cells in the blood of EO771 mice, inducing the immune memory against EO771 tumor cells. Relatively lower dose regimens of 12b1(0.1 µg)/Mix displayed better tumor suppression by more potent STING pathway activation and higher levels of cytokines induction in the tumor.


Assuntos
Citocinas , Neoplasias , Animais , Camundongos , Lipídeos , Nucleotídeos de Citosina/química , Nucleotídeos de Citosina/metabolismo
14.
Carbohydr Polym ; 303: 120477, 2023 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-36657850

RESUMO

The effects of starch granule-associated surface lipids removal on hull-less barley starch structure formed by heat-moisture treatment were investigated. Removing surface lipids made the peak at 2θ of 13° disappear and resulted in higher lamellar peak intensity after harsh treatment and a lower reduction in mass fractal dimension (from 2.49 to 2.43) and radius of gyration (from 24.3 to 24.0) when temperature increased from 100 to 120 °C at 20 % moisture. Treatment at 25 % moisture and 120 °C decreased relative crystallinity (from 15.73 % to 7.43 %) and Gaussian peak area (from 646.7 to 137.7) of native starch, and decreased relative crystallinity (from 14.24 % to 12.56 %) and Gaussian peak area (from 604.1 to 539.6) for starch without surface lipids. Different trends of change in lamellar thickness, linear crystallinity, peak temperatures, and enthalpy of gelatinization were observed among modified starches with increasing temperature and/or moisture content. These results demonstrate that removing surface lipids changes structure of heat-moisture treated starch.


Assuntos
Hordeum , Amido , Amido/química , Temperatura Alta , Temperatura , Lipídeos
15.
J Exp Clin Cancer Res ; 42(1): 7, 2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36604676

RESUMO

BACKGROUND: Triple-Negative Breast Cancer (TNBC) is a subtype of breast cancer that differs from other types of breast cancers in the faster spread and worse outcome. TNBC presented limited treatment options. BET (Bromodomain and extra-terminal domain) proteins are epigenetic readers that control the expression of different oncogenic proteins, and their inhibition (BETi) is considered a promising anti-cancer strategy. Recent evidence demonstrated the involvement of BET proteins in regulation of metabolic processes. METHODS: MDA-MB231 cells treated with JQ1 followed by RNA-sequencing analysis showed altered expression of lipid metabolic genes; among these, we focused on ATGL, a lipase required for efficient mobilization of triglyceride. Different in vitro approaches were performed to validate the RNA-sequencing data (qRT-PCR, immunofluorescence and flow cytometry). NMR (Nuclear Magnetic Resonance) was used to analyze the lipid reprogramming upon treatment. ATGL expression was determined by immunoblot and qRT-PCR, and the impact of ATGL function or protein knockdown, alone and in combination with BETi, was assessed by analyzing cell proliferation, mitochondrial function, and metabolic activity in TNBC and non-TNBC cells culture models. RESULTS: TNBC cells treated with two BETi markedly increased ATGL expression and lipolytic function and decreased intracellular lipid content in a dose and time-dependent manner. The intracellular composition of fatty acids (FAs) after BETi treatment reflected a significant reduction in neutral lipids. The short-chain FA propionate entered directly into the mitochondria mimicking ATGL activity. ATGL KD (knockdown) modulated the levels of SOD1 and CPT1a decreasing ROS and helped to downregulate the expression of mitochondrial ß-oxidation genes in favor of the upregulation of glycolytic markers. The enhanced glycolysis is reflected by the increased of the mitochondrial activity (MTT assay). Finally, we found that after BETi treatment, the FoxO1 protein is upregulated and binds to the PNPLA2 promoter leading to the induction of ATGL. However, FoxO1 only partially prompted the induction of ATGL expression by BETi. CONCLUSIONS: The anti-proliferative effect achieved by BETi is helped by ATGL mediating lipolysis. This study showed that BETi altered the mitochondrial dynamics taking advantage of ATGL function to induce cell cycle arrest and cell death. Schematic representation of BETi mechanism of action on ATGL in TNBC cells. BETi induce the expression of FoxO1 and ATGL, lowering the expression of G0G2, leading to a switch in metabolic status. The induced expression of ATGL leads to increased lipolysis and a decrease in lipid droplet content and bioavailability of neutral lipid. At the same time, the mitochondria are enriched with fatty acids. This cellular status inhibits cell proliferation and increases ROS production and mitochondrial stress. Interfering for ATGL expression, the oxidative phenotypic status mildly reverted to a glycolytic status where neutral lipids are stored into lipid droplets with a consequent reduction of oxidative stress in the mitochondrial.


Assuntos
Aciltransferases , Lipase , Neoplasias de Mama Triplo Negativas , Humanos , Linhagem Celular Tumoral , Ácidos Graxos , Lipase/genética , Lipase/metabolismo , Lipídeos , Proteínas , Espécies Reativas de Oxigênio , Neoplasias de Mama Triplo Negativas/patologia , Aciltransferases/genética , Aciltransferases/metabolismo
16.
Anal Chim Acta ; 1241: 340807, 2023 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-36657877

RESUMO

The metabolome and lipidome are critical components in illustrating biological processes and pathological mechanisms. Generally, two or more independent methods are required to analyze the two compound panels due to their distinct chemical properties and polarity differences. Here, a novel strategy integrating stepwise solid-phase extraction (SPE) and dansyl chemical derivatization was proposed for all-in-one injection LC-MS/MS analysis of serum metabolome and lipidome. In this workflow, a stepwise elution procedure was firstly optimized to separate the metabolome and lipidome fractions using an Ostro plate. Dansyl chemical derivatization was then applied to label amine/phenol, carboxyl, and carbonyl-containing sub-metabolomes. Our results demonstrated that the dansyl labeling could significantly improve chromatographic separation, enhance the MS response, and overcome the matrix effect of co-eluting lipids. Ultimately, an all-in-one injection LC-MS/MS method measuring 256 lipids (covering 20 subclasses) and 212 metabolites (including amino acids, bile acids, fatty acids, acylcarnitines, indole derivatives, ketones and aldehydes, nucleic acid metabolism, polyamines, etc.) was established. This method was applied to investigate the metabolic changes in cisplatin-induced nephrotoxicity in rats and the results were compared with previous untargeted metabolomics. The presented strategy could predominantly improve the analytical coverage and throughput and can be of great use in discovering reliable potential biomarkers in various applications.


Assuntos
Lipidômica , Espectrometria de Massas em Tandem , Animais , Ratos , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Metaboloma , Metabolômica/métodos , Poliaminas/metabolismo , Lipídeos , Extração em Fase Sólida/métodos
17.
Nat Commun ; 14(1): 330, 2023 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-36658137

RESUMO

For volatile organic compounds (VOCs) to be released from the plant cell into the atmosphere, they have to cross the plasma membrane, the cell wall, and the cuticle. However, how these hydrophobic compounds cross the hydrophilic cell wall is largely unknown. Using biochemical and reverse-genetic approaches combined with mathematical simulation, we show that cell-wall localized non-specific lipid transfer proteins (nsLTPs) facilitate VOC emission. Out of three highly expressed nsLTPs in petunia petals, which emit high levels of phenylpropanoid/benzenoid compounds, only PhnsLTP3 contributes to the VOC export across the cell wall to the cuticle. A decrease in PhnsLTP3 expression reduces volatile emission and leads to VOC redistribution with less VOCs reaching the cuticle without affecting their total pools. This intracellular build-up of VOCs lowers their biosynthesis by feedback downregulation of phenylalanine precursor supply to prevent self-intoxication. Overall, these results demonstrate that nsLTPs are intrinsic members of the VOC emission network, which facilitate VOC diffusion across the cell wall.


Assuntos
Compostos Orgânicos Voláteis , Compostos Orgânicos Voláteis/metabolismo , Difusão , Fenilalanina , Parede Celular/metabolismo , Lipídeos
18.
Sci Rep ; 13(1): 1046, 2023 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-36658173

RESUMO

Mule ducks have been force-fed to develop a hepatic steatosis, also called "foie gras", which is similar to the non-alcoholic fatty liver disease (NAFLD) described in humans and mammals. However, in hepatic steatosis resulting from force-feeding of ducks, very little is known about the fine biochemical events that occur due to the enormous and very rapid increase in total lipids that mainly accumulate in hepatocytes. To begin to reduce this lack of knowledge associated with the development of this specific hepatic steatosis, liver samples were taken at different times to follow the overall biochemical transformation of the liver as well as different markers of oxidative stress, hypoxia and apoptosis. The results indicate that the lipid content increases rapidly in the liver throughout the force-feeding period while the protein content decreases. The amount of hydroxyproline remains constant indicating that no liver fibrosis develops during the force-feeding period. On the contrary, all the tested biomarkers of cellular oxidative stress increase rapidly but without any visible disorder in the coordination of paired activities. At the same time, hypoxia-inducible factors also increase indicating that a hypoxia situation is gradually occurring in hepatocytes. This leads, in addition to the lipotoxicity induced by the accumulation of lipids, to an increased number of liver cells to enter into apoptosis. A relative variability in the level of these cellular responses was also observed indicating that, probably, certain animals support the development of this steatosis differently. This leads us to imagine that the physiological status of these birds may differ widely for reasons that remain to be clarified.


Assuntos
Patos , Hepatopatia Gordurosa não Alcoólica , Animais , Humanos , Patos/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Estresse Oxidativo , Hepatócitos/metabolismo , Hipóxia/metabolismo , Lipídeos , Mamíferos
19.
Redox Rep ; 28(1): 2160569, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36661246

RESUMO

PURPOSE: Polycystic ovary syndrome (PCOS) has a series of reproductive and metabolic consequences. Although the link between PCOS, IR, and obesity, their impact on the pathogenesis of PCOS has yet to be determined. Dysfunction of PI3K/AKT pathway has been reported as the main cause of IR in PCOS. This study purposed to explore the effects of selenium nanoparticles (SeNPs) alone and combined with metformin (MET) in a PCOS-IR rat model. METHODS: After 3 weeks of treatment with SeNPs and/or MET, biochemical analysis of glycemic & lipid profiles, and serum reproductive hormones was performed. Inflammatory, oxidative stress, and mitochondrial dysfunction markers were determined colormetrically. The expression of PI3K and Akt genes were evaluated by Real-time PCR. Histopathological examination and Immunohistochemical analysis of Ki-67 expression were performed. RESULTS: The results showed that treatment with SeNPs and/or MET significantly attenuated insulin sensitivity, lipid profile, sex hormones levels, inflammatory, oxidative stress and mitochondrial functions markers. Additionally, PI3K and Akt genes expression were significantly upregulated with improved ovarian histopathological changes. CONCLUSION: Combined SeNPs and MET therapy could be potential therapeutic agent for PCOS-IR model via modulation of the PI3K/Akt pathway, enhancing anti-inflammatory and anti-oxidant properties and altered mitochondrial functions.HighlightsThe strong relationship between obesity, insulin resistance, and polycystic ovarian syndrome.Disturbance of the PI3K/Akt signaling pathway is involved in the progression of polycystic ovary syndrome-insulin resistance (PCOS-IR).In PCOS-IR rats, combined SeNPs and metformin therapy considerably alleviated IR by acting on the PI3K/Akt signaling pathway.The combination of SeNPs and metformin clearly repaired ovarian polycystic pathogenesis and improved hormonal imbalance in PCOS-IR rats.


Assuntos
Resistência à Insulina , Metformina , Nanopartículas , Síndrome do Ovário Policístico , Selênio , Feminino , Humanos , Ratos , Animais , Síndrome do Ovário Policístico/tratamento farmacológico , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/metabolismo , Letrozol/metabolismo , Letrozol/farmacologia , Letrozol/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosfatidilinositol 3-Quinases/farmacologia , Fosfatidilinositol 3-Quinases/uso terapêutico , Selênio/uso terapêutico , Selênio/metabolismo , Selênio/farmacologia , Metformina/uso terapêutico , Metformina/metabolismo , Metformina/farmacologia , Transdução de Sinais , Oxirredução , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Mitocôndrias/metabolismo , Lipídeos
20.
Int J Mol Sci ; 24(2)2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36675224

RESUMO

Even though the application of Next-Generation Sequencing (NGS) has significantly facilitated the identification of disease-associated mutations, the diagnostic rate of rare diseases is still below 50%. This causes a diagnostic odyssey and prevents specific treatment, as well as genetic counseling for further family planning. Increasing the diagnostic rate and reducing the time to diagnosis in children with unclear disease are crucial for a better patient outcome and improvement of quality of life. In many cases, NGS reveals variants of unknown significance (VUS) that need further investigations. The delineation of novel (lipid) biomarkers is not only crucial to prove the pathogenicity of VUS, but provides surrogate parameters for the monitoring of disease progression and therapeutic interventions. Lipids are essential organic compounds in living organisms, serving as building blocks for cellular membranes, energy storage and signaling molecules. Among other disorders, an imbalance in lipid homeostasis can lead to chronic inflammation, vascular dysfunction and neurodegenerative diseases. Therefore, analyzing lipids in biological samples provides great insight into the underlying functional role of lipids in healthy and disease statuses. The method of choice for lipid analysis and/or huge assemblies of lipids (=lipidome) is mass spectrometry due to its high sensitivity and specificity. Due to the inherent chemical complexity of the lipidome and the consequent challenges associated with analyzing it, progress in the field of lipidomics has lagged behind other omics disciplines. However, compared to the previous decade, the output of publications on lipidomics has increased more than 17-fold within the last decade and has, therefore, become one of the fastest-growing research fields. Combining multiple omics approaches will provide a unique and efficient tool for determining pathogenicity of VUS at the functional level, and thereby identifying rare, as well as novel, genetic disorders by molecular techniques and biochemical analyses.


Assuntos
Lipidômica , Doenças Metabólicas , Criança , Humanos , Doenças Raras/diagnóstico , Doenças Raras/genética , Lipídeos/química , Medicina de Precisão , Qualidade de Vida , Metabolismo dos Lipídeos , Doenças Metabólicas/diagnóstico , Doenças Metabólicas/genética , Doenças Metabólicas/terapia
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