RESUMO
PURPOSE OF REVIEW: Oxidized low-density lipoprotein (Ox-LDL) and chylomicron remnants were previously proposed as the most atherogenic lipoproteins for the causal lipoproteins of atherosclerosis. However, there are still controversies on these hypothesizes. Therefore, we have proposed a new hypothesis based on our recent findings of remnant lipoproteins (RLPs) in postprandial plasma. RECENT FINDINGS: Plasma RLP-C and RLP-TG increased significantly after fat load. More than 80% of the increased triglycerides after fat load consisted of the triglycerides in RLP, which contained greater amount of apoB100 than apoB48 particles as mostly very low density lipoproteins (VLDL) remnants. The majority of lipoprotein lipase (LPL) in plasma was found in RLP as RLP-LPL complex, which is released into circulation after hydrolysis. LPL activity and concentration in plasma did not increase after food intake associated with the insufficient hydrolysis of chylomicrons and VLDL and resulted in the significant increase of RLP-TG. Plasma LPL was inversely correlated with RLP particle size and number. SUMMARY: VLDL remnants have been shown as the major atherogenic lipoproteins in postprandial plasma associated with LPL activity as the targets for prevention of atherosclerosis. We also proposed a new definition of RLPs, 'LPL bound TG-rich lipoproteins' based on the findings of RLP-LPL complex.
Assuntos
Aterosclerose/prevenção & controle , Remanescentes de Quilomícrons/fisiologia , Hipolipemiantes/uso terapêutico , Lipoproteínas VLDL/fisiologia , Lipoproteínas/sangue , Terapia de Alvo Molecular/métodos , Aterosclerose/sangue , Remanescentes de Quilomícrons/antagonistas & inibidores , Humanos , Hiperlipidemias/sangue , Hiperlipidemias/complicações , Hipertrigliceridemia/sangue , Hipertrigliceridemia/complicações , Lipoproteínas/antagonistas & inibidores , Lipoproteínas LDL , Lipoproteínas VLDL/antagonistas & inibidores , Período Pós-Prandial/fisiologia , Triglicerídeos/sangueRESUMO
SCOPE: This study aims to characterize the effect of fenugreek (Trigonella foenum-graecum) seed and its phytoceutical trigonelline in antimetabolic inflammation and ameliorating overproduction of very low density lipoprotein (VLDL) in insulin resistance. METHODS AND RESULTS: Two groups of genetic hyperlipidemic mice generated by depletion of cAMP responsive element binding protein H (CREBH) are fed either a chow containing 2% fenugreek seed or vehicle for 7 weeks. Q-RT-PCR and immunoblotting analysis demonstrated that fenugreek seed containing diet inhibits hepatic SREBP-1c activation and the subsequent de novo lipogenesis by enhancing expression of insulin-inducible gene-1 (Insig-1) and gene-2 (Insig-2). mRNA expression of PPARα and its target genes that are involved in fatty acid ß-oxidation are also upregulated in the fenugreek seed fed-mice which is accompanied by significantly reduced hepatic lipid accumulation and VLDL secretion, improved endoplasmic reticulum (ER) stress, and ameliorated metabolic inflammation. These actions enhance insulin sensitivity and improve hyperlipidemia. In vitro, treating a rat hepatoma cell line, McA-RH7777 (McA), with trigonelline is able to recapitulate the results observed in vivo. CONCLUSIONS: This study unveils a novel mechanism of fenugreek seed and trigonelline in countering hepatic VLDL overproduction and insulin resistance by enhancing the Insig signaling pathways and ameliorating metabolic inflammatory stress in the liver.
Assuntos
Alcaloides/farmacologia , Resistência à Insulina , Lipoproteínas VLDL/antagonistas & inibidores , Proteínas de Membrana/fisiologia , Extratos Vegetais/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Lipogênese/efeitos dos fármacos , Lipoproteínas VLDL/biossíntese , Camundongos , Ratos , Transdução de Sinais/fisiologia , Proteína de Ligação a Elemento Regulador de Esterol 1/fisiologia , Trigonella , Células Tumorais CultivadasRESUMO
Our previous studies demonstrated that the first 1000 amino acid residues (the ßα1 domain) of human apolipoprotein (apo) B-100, termed apoB:1000, are required for the initiation of lipoprotein assembly and the formation of a monodisperse stable phospholipid (PL)-rich particle. The objectives of this study were (a) to assess the effects on the properties of apoB truncates undergoing sequential inclusion of the amphipathic ß strands in the 700 N-terminal residues of the ß1 domain of apoB-100 and (b) to identify the subdomain in the ß1 domain that is required for the formation of a microsomal triglyceride transfer protein (MTP)-dependent triacylglycerol (TAG)-rich apoB-containing particle. Characterization of particles secreted by stable transformants of McA-RH7777 cells demonstrated the following. (1) The presence of amphipathic ß strands in the 200 N-terminal residues of the ß1 domain resulted in the secretion of apoB truncates (apoB:1050 to apoB:1200) as both lipidated and lipid-poor particles. (2) Inclusion of residues 300-700 of the ß1 domain led to the secretion of apoB:1300, apoB:1400, apoB:1500, and apoB:1700 predominantly as lipidated particles. (3) Particles containing residues 1050-1500 were all rich in PL. (4) There was a marked increase in the lipid loading capacity and TAG content of apoB:1700-containing particles. (5) Only the level of secretion of apoB:1700 was markedly diminished by MTP inhibitor BMS-197636. These results suggest that apoB:1700 marks the threshold for the formation of a TAG-rich particle and support the concept that MTP participates in apoB assembly and secretion at the stage where particles undergo a transition from PL-rich to TAG-rich.
Assuntos
Apolipoproteína B-100/química , Proteínas de Transporte/metabolismo , Hepatócitos/metabolismo , Lipoproteínas VLDL/metabolismo , Animais , Apolipoproteína B-100/genética , Apolipoproteína B-100/metabolismo , Transporte Biológico/efeitos dos fármacos , Proteínas de Transporte/antagonistas & inibidores , Linhagem Celular Tumoral , Fluorenos/farmacologia , Hepatócitos/efeitos dos fármacos , Humanos , Isoindóis/farmacologia , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/química , Peso Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Fosfolipídeos/análise , Fosfolipídeos/metabolismo , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Dobramento de Proteína , Domínios e Motivos de Interação entre Proteínas , Estabilidade Proteica , Proteólise/efeitos dos fármacos , Ratos , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo , Triglicerídeos/análise , Triglicerídeos/metabolismoRESUMO
CONTEXT: Growth hormone (GH) stimulates hepatic synthesis of very-low-density lipoproteins (VLDL), whereas hepatic steatosis develops as a result of GH deficiency. Steatosis is also a complication of tamoxifen treatment, the cause of which is not known. As tamoxifen inhibits the secretion and action of GH, we hypothesize that it induces steatosis by inhibiting hepatic VLDL export. AIM: To investigate whether tamoxifen reduces hepatic VLDL secretion. DESIGN: Eight healthy, normolipidemic women (age: 64.4 ± 2.1 years) were studied in random sequence at baseline, after 2 weeks of tamoxifen (20 mg/day) and after 2 weeks of estradiol valerate (EV; 2 mg/day) treatments, separated by a 4-week washout period. The kinetics of apolipoprotein B (apoB), the structural protein of VLDL particles, were measured using a stable isotope 2H3-leucine turnover technique. VLDL-apoB fractional catabolic rate (FCR) was determined using a multicompartment model. VLDL-apoB secretion was estimated as the product of FCR and VLDL-apoB concentration. GH response to arginine stimulation, circulating levels of IGF-1, FFA, and TG, along with TG content in VLDL were measured. RESULTS: Tamoxifen significantly (P < 0.05) reduced VLDL-apoB concentration and secretion by 27.3 ± 7.8% and 29.8 ± 10.2%, respectively. In contrast, EV did not significantly change VLDL-apoB concentration or secretion. Tamoxifen but not EV significantly reduced (P < 0.05) GH response to arginine stimulation. Both treatments significantly lowered (P < 0.05) circulating IGF-1. CONCLUSION: Inhibition of VLDL secretion may contribute to the development of fatty liver during tamoxifen therapy. As GH stimulates VLDL secretion, the development of steatosis may arise secondarily from GH insufficiency induced by tamoxifen.
Assuntos
Fígado Gorduroso/sangue , Fígado Gorduroso/induzido quimicamente , Hormônio do Crescimento Humano/sangue , Lipoproteínas VLDL/sangue , Fígado/metabolismo , Tamoxifeno/efeitos adversos , Idoso , Antagonistas de Estrogênios/efeitos adversos , Antagonistas de Estrogênios/farmacologia , Fígado Gorduroso/diagnóstico , Feminino , Hormônio do Crescimento Humano/antagonistas & inibidores , Humanos , Lipoproteínas VLDL/antagonistas & inibidores , Fígado/efeitos dos fármacos , Pessoa de Meia-Idade , Tamoxifeno/farmacologiaRESUMO
Hypertriglyceridemia (triglycerides > 150 mg/dL) affects ~25 % of the United States (US) population and is associated with increased cardiovascular risk. Severe hypertriglyceridemia (≥ 500 mg/dL) is also a risk factor for pancreatitis. Three omega-3 fatty acid (OM3FA) prescription formulations are approved in the US for the treatment of adults with severe hypertriglyceridemia: (1) OM3FA ethyl esters (OM3EE), a mixture of OM3FA ethyl esters, primarily eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (Lovaza®, Omtryg™, and generics); (2) icosapent ethyl (IPE), EPA ethyl esters (Vascepa®); and (3) omega-3 carboxylic acids (OM3CA), a mixture of OM3FAs in free fatty acid form, primarily EPA, DHA, and docosapentaenoic acid (Epanova®). At approved doses, all formulations substantially reduce triglyceride and very-low-density lipoprotein levels. DHA-containing formulations may also increase low-density lipoprotein cholesterol. However, this is not accompanied by increased non-high-density lipoprotein cholesterol, which is thought to provide a better indication of cardiovascular risk in this patient population. Proposed mechanisms of action of OM3FAs include inhibition of diacylglycerol acyltransferase, increased plasma lipoprotein lipase activity, decreased hepatic lipogenesis, and increased hepatic ß-oxidation. OM3CA bioavailability (area under the plasma concentration-time curve from zero to the last measurable concentration) is up to 4-fold greater than that of OM3FA ethyl esters, and unlike ethyl esters, the absorption of OM3CA is not dependent on pancreatic lipase hydrolysis. All three formulations are well tolerated (the most common adverse events are gastrointestinal) and demonstrate a lack of drug-drug interactions with other lipid-lowering drugs, such as statins and fibrates. OM3FAs appear to be an effective treatment option for patients with severe hypertriglyceridemia.
Assuntos
Ácidos Docosa-Hexaenoicos/uso terapêutico , Ácido Eicosapentaenoico/análogos & derivados , Ácidos Graxos Ômega-3/uso terapêutico , Hipertrigliceridemia/tratamento farmacológico , Hipolipemiantes/uso terapêutico , Triglicerídeos/antagonistas & inibidores , Adulto , Disponibilidade Biológica , LDL-Colesterol/sangue , Diacilglicerol O-Aciltransferase/antagonistas & inibidores , Diacilglicerol O-Aciltransferase/metabolismo , Ácidos Docosa-Hexaenoicos/farmacocinética , Combinação de Medicamentos , Ácido Eicosapentaenoico/farmacocinética , Ácido Eicosapentaenoico/uso terapêutico , Ésteres , Ácidos Graxos Ômega-3/farmacocinética , Humanos , Hipertrigliceridemia/sangue , Hipertrigliceridemia/patologia , Hipolipemiantes/farmacocinética , Lipogênese/efeitos dos fármacos , Lipase Lipoproteica/metabolismo , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/sangue , Fígado/efeitos dos fármacos , Fígado/metabolismo , Oxirredução/efeitos dos fármacos , Triglicerídeos/sangueRESUMO
OBJECTIVE: We aimed to evaluate the effect of atorvastatin on apolipoprotein AV (ApoAV) in HepG2 cells of insulin resistance (IR), and further explore its mechanism. MATERIALS AND METHODS: Firstly, a model of IR in HepG2 cells was established by insulin, and then treated with various concentrations of atorvastatin (0, 10, 100 and 500 nM) for 12 h and 24 h, respectively. Detection of glucose concentration was performed by Glucose Oxidase kit. Subsequently, Enzyme-linked immunosorbent assay (ELISA) kits were used to measure the concentrations of triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL) and very low density lipoprotein (VLDL). The mRNA levels of ApoAV and ApoAV-related genes, including glucose transporter 1 (Glut1), Glut2, peroxisome proliferator activated receptor α (PPARα), and liver X receptor α (LXRα) were detected by qRT-PCR. RESULTS: We successfully established IR model in HepG2 cells by 10-6 nM insulin. Subsequently, we found that the glucose extraction rate and mRNA level of ApoAV significantly reduced in HepG2 cells of IR (p < 0.05); however, atorvastatin increased the glucose extraction rate and ApoAV mRNA level. Furthermore, atorvastatin inhibited the concentration of TG in HepG2 cells of IR (p < 0.05); however, atorvastatin had no effect on HDL, LDL and VLDL. Also, atorvastatin could increase the mRNA levels of Glut2 but not Glut1, PPARα, and LXRα. CONCLUSIONS: Our study indicated that atorvastatin might inhibit IR induced by insulin through the TG-lowering role of ApoAV. Furthermore, Glut2 might be involved in the effect of atorvastatin on ApoAV in HepG2 cells of IR.
Assuntos
Anticolesterolemiantes/farmacologia , Apolipoproteínas A/biossíntese , Atorvastatina/farmacologia , Resistência à Insulina/fisiologia , Insulina/toxicidade , Triglicerídeos/metabolismo , Apolipoproteína A-V , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Células Hep G2 , Humanos , Lipoproteínas HDL/antagonistas & inibidores , Lipoproteínas HDL/metabolismo , Lipoproteínas LDL/antagonistas & inibidores , Lipoproteínas LDL/metabolismo , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/metabolismo , Triglicerídeos/antagonistas & inibidoresRESUMO
Familial hypercholesterolemia (FH) is a genetic disorder characterized by extremely high levels of plasma low-density lipoprotein (LDL), due to defective LDL receptor-apolipoprotein B (APOB) binding. Current therapies such as statins or LDL apheresis for homozygous FH are insufficiently efficacious at lowering LDL cholesterol or are expensive. Treatments that target APOB100, the structural protein of LDL particles, are potential therapies for FH. We have developed a series of APOB-directed splice-switching oligonucleotides (SSOs) that cause the expression of APOB87, a truncated isoform of APOB100. APOB87, like similarly truncated isoforms expressed in patients with a different condition, familial hypobetalipoproteinemia, lowers LDL cholesterol by inhibiting very low-density lipoprotein (VLDL) assembly and increasing LDL clearance. We demonstrate that these "APO-skip " SSOs induce high levels of exon skipping and expression of the APOB87 isoform, but do not substantially inhibit APOB48 expression in cell lines. A single injection of an optimized APO-skip SSO into mice transgenic for human APOB resulted in abundant exon skipping that persists for >6 days. Weekly treatments generated a sustained reduction in LDL cholesterol levels of 34-51% in these mice, superior to pravastatin in a head-to-head comparison. These results validate APO-skip SSOs as a candidate therapy for FH.
Assuntos
Apolipoproteínas B/genética , LDL-Colesterol/sangue , Éxons , Oligonucleotídeos/genética , Precursores de RNA/genética , Splicing de RNA , Animais , Apolipoproteínas B/metabolismo , Células CACO-2 , Terapia Genética/métodos , Células Hep G2 , Homozigoto , Humanos , Hiperlipoproteinemia Tipo II/sangue , Hiperlipoproteinemia Tipo II/genética , Hiperlipoproteinemia Tipo II/terapia , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/sangue , Fígado/metabolismo , Camundongos , Camundongos Transgênicos , Oligonucleotídeos/metabolismo , Precursores de RNA/metabolismo , Coelhos , Receptores de Lipoproteínas/genética , Receptores de Lipoproteínas/metabolismo , Análise de Sequência de RNARESUMO
Insulin suppresses secretion of very low density lipoprotein (VLDL) apolipoprotein (apo) B in primary rodent hepatocytes (RH) by favoring the degradation of B100, the larger form of apo B, through post-endoplasmic reticulum proteolysis. Sortilin 1 (sort1), a multi-ligand sorting receptor, has been proposed as a mediator of lysosomal B100 degradation by directing B100 in pre-VLDL to lysosomes rather than allowing maturation to VLDL and secretion. The purpose of our studies was to investigate the role of sort1 in insulin-dependent degradation of apo B. Using liver derived McArdle RH7777 (McA) cells, we demonstrate that insulin suppresses VLDL B100 secretion via a phosphatidylinositide 3-kinase (PI3K) dependent process that is inhibitable by wortmannin in a fashion similar to RH. Using McA cells and in situ cross-linking, we demonstrate that insulin acutely (30min) stimulates the interaction of B100 with sort1. The insulin-induced interaction of sort1-B100 is markedly enhanced when lysosomal degradation is inhibited by Bafilomycin A1 (BafA1), an inhibitor of lysosomal acidification. As BafA1 also prevents insulin suppressive effects on apo B secretion, our results suggest that sort1-B100 interaction stimulated by insulin transiently accumulates with BafA1 and favors B100 secretion by default.
Assuntos
Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Apolipoproteínas B/antagonistas & inibidores , Hepatócitos/efeitos dos fármacos , Insulina/farmacologia , Lisossomos/efeitos dos fármacos , Animais , Apolipoproteínas B/metabolismo , Linhagem Celular , Hepatócitos/metabolismo , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/metabolismo , Lisossomos/metabolismo , Macrolídeos/farmacologia , RatosRESUMO
Microsomal triglyceride transfer protein (MTP) is involved in the synthesis of very low density lipoprotein in the liver. Its deficiency results in abetalipoproteinemia. MTP inhibitors target the assembly and secretion of apolipoprotein B-containing lipoproteins. These agents may potentially play a role, alone or in combination, in the treatment of hypercholesterolemia or hypertriglyceridaemia. Clinical applications of MTP inhibitors initially focused primarily on high-dose monotherapy in order to produce substantial reductions in LDL-cholesterol levels but these proved to induce significant hepatic steatosis and transaminase elevations. However, likely orphan indications for MTP inhibitors, where a different risk-benefit profile applies, include patients with homozygous familial hypercholesterolemia where statins often show a low response. Development of MTP inhibitors has continued to enter clinical trials at lower doses or in formulations aimed at utilizing their efficacy while avoiding their side effects. These have shown promising results in reducing cholesterol, triglycerides and apolipoprotein B with a far lower incidence of, often, transient side-effects. The clinical efficacy and safety of MTP inhibition in patients with hyperlipidaemia remains to be fully determined and to be proven in both surrogate and clinical endpoint trials but there may be a role for these agents in orphan indications for rarer severe hyperlipidaemias.
Assuntos
Abetalipoproteinemia/tratamento farmacológico , Apolipoproteínas B/antagonistas & inibidores , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/metabolismo , Hipercolesterolemia/tratamento farmacológico , Hiperlipoproteinemia Tipo II/tratamento farmacológico , Hipertrigliceridemia/tratamento farmacológico , Abetalipoproteinemia/metabolismo , Apolipoproteínas B/metabolismo , Humanos , Hipercolesterolemia/metabolismo , Hiperlipoproteinemia Tipo II/metabolismo , Hipertrigliceridemia/metabolismo , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/metabolismo , Microssomos/efeitos dos fármacos , Microssomos/fisiologiaRESUMO
VLDL secretion is a regulated process that depends on the availability of lipids, apoB and MTP. Our aim was to investigate the effect of liver denervation upon the secretion of VLDL and the expression of proteins involved in this process. Denervation was achieved by applying a 85% phenol solution onto the portal tract, while control animals were treated with 9% NaCl. VLDL secretion was evaluated by the Tyloxapol method. The hepatic concentration of TAG and cholesterol, and the plasma concentration of TAG, cholesterol, VLDL-TAG, VLDL-cholesterol and HDL-cholesterol were measured, as well as mRNA expression of proteins involved in the process of VLDL assembly. Hepatic acinar distribution of MTP and apoB was evaluated by immunohistochemistry. Denervation increased plasma concentration of cholesterol (125.3 +/- 10.1 vs. 67.1 +/- 4.9 mg dL(-1)) and VLDL-cholesterol (61.6 +/- 5.6 vs. 29.4 +/- 3.3 mg dL(-1)), but HDL-cholesterol was unchanged (45.5 +/- 6.1 vs. 36.9 +/- 3.9 mg dL(-1)). Secretion of VLDL-TAG (47.5 +/- 23.8 vs. 148.5 +/- 27.4 mg dL h(-1)) and mRNA expression of CPT I and apoB were reduced (p < 0.01) in the denervated animals. MTP and apoB acinar distribution was not altered in the denervated animals, but the intensity of the reaction was reduced in relation to controls.
Assuntos
Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/biossíntese , Fígado/inervação , Fígado/metabolismo , Triglicerídeos/antagonistas & inibidores , Triglicerídeos/biossíntese , Animais , Apolipoproteínas B/metabolismo , Colesterol/sangue , Denervação , Lipoproteínas VLDL/sangue , Lipoproteínas VLDL/metabolismo , Masculino , Ratos , Ratos Wistar , Triglicerídeos/sangue , Triglicerídeos/metabolismoRESUMO
AIMS/HYPOTHESIS: Overproduction of VLDL(1) seems to be the central pathophysiological feature of the dyslipidaemia associated with type 2 diabetes. We explored the relationship between liver fat and suppression of VLDL(1) production by insulin in participants with a broad range of liver fat content. METHODS: A multicompartmental model was used to determine the kinetic parameters of apolipoprotein B and TG in VLDL(1) and VLDL(2) after a bolus of [(2)H(3)]leucine and [(2)H(5)]glycerol during a hyperinsulinaemic-euglycaemic clamp in 20 male participants: eight with type 2 diabetes and 12 control volunteers. The participants were divided into two groups with low or high liver fat. All participants with diabetes were in the high liver-fat group. RESULTS: The results showed a rapid drop in VLDL(1)-apolipoprotein B and -triacylglycerol secretion in participants with low liver fat during the insulin infusion. In contrast, participants with high liver fat showed no significant change in VLDL(1) secretion. The VLDL(1) suppression following insulin infusion correlated with the suppression of NEFA, and the ability of insulin to suppress the plasma NEFA was impaired in participants with high liver fat. A novel finding was an inverse response between VLDL(1) and VLDL(2) secretion in participants with low liver fat: VLDL(1) secretion decreased acutely after insulin infusion whereas VLDL(2) secretion increased. CONCLUSIONS/INTERPRETATION: Insulin downregulates VLDL(1) secretion and increases VLDL(2) secretion in participants with low liver fat but fails to suppress VLDL(1) secretion in participants with high liver fat, resulting in overproduction of VLDL(1). Thus, liver fat is associated with lack of VLDL(1) suppression in response to insulin.
Assuntos
Tecido Adiposo/anatomia & histologia , Diabetes Mellitus Tipo 2/fisiopatologia , Resistência à Insulina/fisiologia , Insulina/fisiologia , Lipoproteínas VLDL/metabolismo , Fígado/anatomia & histologia , Abdome , Tecido Adiposo/fisiologia , Adulto , Apolipoproteínas B/sangue , Índice de Massa Corporal , Diabetes Mellitus Tipo 2/sangue , Ácidos Graxos não Esterificados/metabolismo , Humanos , Insulina/sangue , Cinética , Lipoproteínas VLDL/antagonistas & inibidores , Fígado/metabolismo , Masculino , Pessoa de Meia-Idade , Valores de ReferênciaRESUMO
We used gene knockout mice to explore the role of Angiopoietin-like-4 (Angptl4) in lipid metabolism as well as to generate anti-Angptl4 mAbs with pharmacological activity. Angptl4 -/- mice had lower triglyceride (TG) levels resulting both from increased very low-density lipoprotein (VLDL) clearance and decreased VLDL production and had modestly lower cholesterol levels. Also, both Angptl4 -/- suckling mice and adult mice fed a high-fat diet showed reduced viability associated with lipogranulomatous lesions of the intestines and their draining lymphatics and mesenteric lymph nodes. Treating C57BL/6J, ApoE -/-, LDLr -/-, and db/db mice with the anti-Angptl4 mAb 14D12 recapitulated the lipid and histopathologic phenotypes noted in Angptl4 -/- mice. This demonstrates that the knockout phenotype reflects not only the physiologic function of the Angptl4 gene but also predicts the pharmacologic consequences of Angptl4 protein inhibition with a neutralizing antibody in relevant models of human disease.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Proteínas Sanguíneas/genética , Proteínas Sanguíneas/imunologia , Hipolipemiantes/administração & dosagem , Lipídeos/antagonistas & inibidores , Fenótipo , Proteína 4 Semelhante a Angiopoietina , Angiopoietinas , Animais , Anticorpos Bloqueadores/administração & dosagem , Anticorpos Bloqueadores/biossíntese , Anticorpos Monoclonais/biossíntese , Proteínas Sanguíneas/deficiência , Proteínas Sanguíneas/fisiologia , Hipolipemiantes/metabolismo , Hipolipemiantes/uso terapêutico , Lipídeos/biossíntese , Lipídeos/sangue , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/sangue , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Triglicerídeos/antagonistas & inibidores , Triglicerídeos/sangueRESUMO
Ferrylmyoglobin (ferrylMb) may play a major role in vivo under certain pathological conditions. Preliminary experiments showed that ferrylmyoglobin induced a mild oxidative stress in rat hepatocytes, mainly reflected by early lipid peroxidation. One of the major functions of hepatocytes is the synthesis, secretion and distribution of lipids to other cells. The aim of this work was to examine whether ferrylMb affected the synthesis and secretion of triacylglycerols (TAG), and the possible involvement of lipid peroxidation on these effects. The heme protein completely impaired VLDL secretion, affecting both the lipid and apoB components of the lipoprotein particle. The incorporation of [(3)H]-oleate into newly synthesized diacylglycerol and TAG was not altered by ferrylMb. The co-treatment of cells with alpha-tocopherol prevented lipid peroxidation and concomitantly reverted VLDL TAG secretion to control values. Importantly, although ferrylMb dramatically blocked prelabeled TAG secretion, newly synthesized TAG secretion was not impaired. These data indicate that lipid peroxidation elicited by ferrylMb modulates the VLDL TAG secretion process, specifically affecting the stored intracellular TAG mobilization, rather than de novo synthesis. Apart from its potential role in vivo, ferrylmyoglobin constitutes a useful model for studying the interactions between lipid peroxidation and the specific TAG pool dependence for VLDL secretion.
Assuntos
Peroxidação de Lipídeos/fisiologia , Lipoproteínas VLDL/metabolismo , Fígado/fisiologia , Metamioglobina/farmacologia , Triglicerídeos/metabolismo , Animais , Apolipoproteína B-100/metabolismo , Diglicerídeos/metabolismo , Cinética , Peroxidação de Lipídeos/efeitos dos fármacos , Lipídeos/isolamento & purificação , Lipoproteínas VLDL/antagonistas & inibidores , Fígado/efeitos dos fármacos , Masculino , Ratos , Ratos Sprague-Dawley , Transferrina/metabolismo , Triglicerídeos/antagonistas & inibidoresRESUMO
OBJECTIVE: Cholesteryl ester transfer protein (CETP) inhibition with torcetrapib not only increases high-density lipoprotein cholesterol levels but also significantly reduces plasma triglyceride, low-density lipoprotein (LDL) cholesterol, and apolipoprotein B (apoB) levels. The goal of the present study was to define the kinetic mechanism(s) by which CETP inhibition reduces levels of apoB-containing lipoproteins. METHODS AND RESULTS: Nineteen subjects, 9 of whom were pretreated with 20 mg atorvastatin, received placebo for 4 weeks, followed by 120 mg torcetrapib once daily for 4 weeks. Six subjects in the nonatorvastatin group received 120 mg torcetrapib twice daily for an additional 4 weeks. After each phase, subjects underwent a primed-constant infusion of deuterated leucine to endogenously label newly synthesized apoB to determine very low-density lipoprotein (VLDL), intermediate-density lipoprotein (IDL) and LDL apoB100 production, and fractional catabolic rates (FCRs). Once-daily 120 mg torcetrapib significantly reduced VLDL, IDL, and LDL apoB100 pool sizes by enhancing the FCR of apoB100 within each fraction. On a background of atorvastatin, 120 mg torcetrapib significantly reduced VLDL, IDL, and LDL apoB100 pool sizes. The reduction in VLDL apoB100 was associated with an enhanced apoB100 FCR, whereas the decreases in IDL and LDL apoB100 were associated with reduced apoB100 production. CONCLUSIONS: These data indicate that when used alone, torcetrapib reduces VLDL, IDL, and LDL apoB100 levels primarily by increasing the rate of apoB100 clearance. In contrast, when added to atorvastatin treatment, torcetrapib reduces apoB100 levels mainly by enhancing VLDL apoB100 clearance and reducing production of IDL and LDL apoB100.
Assuntos
Apolipoproteínas B/antagonistas & inibidores , Proteínas de Transporte/antagonistas & inibidores , Glicoproteínas/antagonistas & inibidores , Quinolinas/farmacologia , Adulto , Idoso , Apolipoproteína B-100 , Apolipoproteínas B/sangue , Apolipoproteínas B/metabolismo , Atorvastatina , Proteínas de Transferência de Ésteres de Colesterol , Estudos Cross-Over , Sinergismo Farmacológico , Ácidos Heptanoicos/farmacologia , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Cinética , Lipídeos/sangue , Lipoproteínas/antagonistas & inibidores , Lipoproteínas/biossíntese , Lipoproteínas/sangue , Lipoproteínas IDL , Lipoproteínas LDL/antagonistas & inibidores , Lipoproteínas LDL/biossíntese , Lipoproteínas LDL/sangue , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/sangue , Lipoproteínas VLDL/metabolismo , Pessoa de Meia-Idade , Pirróis/farmacologia , Método Simples-CegoRESUMO
OBJECTIVE: We investigated the role of adipocyte differentiation-related protein (ADRP) in triglyceride turnover and in the secretion of very low-density lipoprotein (VLDL) from McA-RH7777 cells and primary rat hepatocytes. METHODS AND RESULTS: An increase in the expression of ADRP increased triglyceride accumulation in cytosolic lipid droplets and prevented the incorporation of fatty acids into secretable triglycerides, thereby reducing the secretion of triglycerides as well as of apolipoprotein B-100 (apoB-100) and apoB-48 VLDL. The ability of ADRP to block the secretion of apoB-100 VLDL1 decreased with increasing quantities of fatty acids in the medium, indicating a saturable process and emphasizing the importance of sequestering of fatty acids for the effect of ADRP on VLDL secretion. Knockdown (small interfering RNA) of ADRP decreased the pool of cytosolic lipid droplets but increased only the secretion of apoB-48 VLDL1. Additionally, there was an increased flow of fatty acids into beta-oxidation. CONCLUSIONS: ADRP is essential for the accumulation of triglycerides in cytosolic lipid droplets. An increase in ADRP prevents the formation of VLDL by diverting fatty acids from the VLDL assembly pathway into cytosolic triglycerides, whereas a decrease of the protein increases the sorting of fatty acids to beta-oxidation and promotes the secretion of apoB-48 VLDL1.
Assuntos
Citosol/metabolismo , Ácidos Graxos/metabolismo , Lipoproteínas VLDL/antagonistas & inibidores , Proteínas de Membrana/fisiologia , Triglicerídeos/metabolismo , Animais , Apolipoproteína B-48 , Apolipoproteínas B/metabolismo , Linhagem Celular Tumoral , Técnicas de Transferência de Genes , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipoproteínas VLDL/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/farmacologia , Oxirredução/efeitos dos fármacos , Perilipina-2 , RNA Interferente Pequeno/farmacologia , RatosRESUMO
The objective of this study was to determine the effect of two amino-terminal apolipoprotein A-I (apoA-I) deletions on high-density lipoprotein (HDL) biosynthesis and lipid homeostasis. Adenovirus-mediated gene transfer showed that the apoA-I[Delta(89-99)] deletion mutant caused hypercholesterolemia, characterized by increased plasma cholesterol and phospholipids, that were distributed in the very low density/intermediate density/low-density lipoprotein (VLDL/IDL/LDL) region, and normal triglycerides. The capacity of the mutant protein to promote ATP-binding cassette transporter A1- (ABCA1-) mediated cholesterol efflux and to activate lecithin:cholesterol acyltranserase (LCAT) was approximately 70-80% of the wild-type (WT) control. The phospholipid transfer protein (PLTP) activity of plasma containing the apoA-I[Delta(89-99)] mutant was decreased to 32% of the WT control. Similar analysis showed that the apoA-I[Delta(62-78)] deletion mutant in apoA-I-deficient mice caused combined hyperlipidemia characterized by increased triglycerides, cholesterol, and phospholipids in the VLDL/IDL region. There was enrichment of the VLDL/IDL with mutant apoA-I that resulted in reduction of in vitro lipolysis. The capacity of this mutant to promote ABCA1-mediated cholesterol efflux was normal, and the capacity to activate LCAT in vitro was reduced by 53%. The WT apoA-I and the apoA-I[Delta(62-78)] mutant formed spherical HDL particles, whereas the apoA-I[Delta(89-99)] mutant formed discoidal HDL particles. We conclude that alterations in apoA-I not only may have adverse effects on HDL biosynthesis but also may promote dyslipidemia due to interference of the apoA-I mutants on the overall cholesterol and triglycerides homeostasis.
Assuntos
Adenoviridae/genética , Apolipoproteína A-I/deficiência , Apolipoproteína A-I/genética , Técnicas de Transferência de Genes , Hipercolesterolemia/genética , Deleção de Sequência , Animais , Apolipoproteína A-I/biossíntese , Apolipoproteína A-I/sangue , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Humanos , Hidrólise , Hipercolesterolemia/sangue , Lipoproteínas/antagonistas & inibidores , Lipoproteínas/sangue , Lipoproteínas HDL/sangue , Lipoproteínas HDL/metabolismo , Lipoproteínas HDL/ultraestrutura , Lipoproteínas IDL , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/sangue , Fígado/metabolismo , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/sangue , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fragmentos de Peptídeos/sangue , Fragmentos de Peptídeos/genética , Proteínas de Transferência de Fosfolipídeos/antagonistas & inibidores , Proteínas de Transferência de Fosfolipídeos/sangue , Estrutura Secundária de Proteína/genética , RNA Mensageiro/biossíntese , Triglicerídeos/antagonistas & inibidores , Triglicerídeos/sangueRESUMO
Recent evidence demonstrates that hypothalamic insulin signaling is required for inhibition of endogenous glucose production. The downstream mechanisms that are responsible for the effects of hypothalamic insulin receptor activation on hepatic fuel flux remain to be determined. To establish whether downregulation of neuropeptide Y (NPY) release by insulin is mandatory for its capacity to suppress glucose production, we examined the effects of a continuous intracerebroventricular (ICV) infusion of NPY (10 microg/h for 3-5 h) on glucose flux during a hyperinsulinemic-euglycemic clamp in mice. We also evaluated the effects of ICV NPY administration on free fatty acid and glycerol flux and VLDL production in this experimental context. In basal conditions, none of the metabolic parameters was affected by NPY infusion. In hyperinsulinemic conditions, peripheral glucose disposal was not different between vehicle- and NPY-infused animals. In contrast, hyperinsulinemia suppressed endogenous glucose production by approximately 8% vs. 30% in NPY- vs. vehicle-infused mice, respectively (P < 0.05). Also, VLDL production was significantly higher during hyperinsulinemia in NPY- compared with vehicle-infused mice (97.5 +/- 18.0 vs. 54.7 +/- 14.9 micromol. kg(-1). h(-1); P < 0.01). These data suggest that the neurophysiological action of insulin to downregulate hypothalamic NPY release is a prerequisite for its ability to suppress hepatic fuel production, whereas it is not mandatory for its capacity to modulate glucose disposal or lipolysis.
Assuntos
Ventrículos Cerebrais/fisiologia , Glucose/biossíntese , Insulina/farmacologia , Lipoproteínas VLDL/biossíntese , Neuropeptídeo Y/farmacologia , Animais , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Ventrículos Cerebrais/efeitos dos fármacos , Ácidos Graxos não Esterificados/sangue , Glucose/antagonistas & inibidores , Técnica Clamp de Glucose , Glicerol/sangue , Hiperinsulinismo/sangue , Infusões Parenterais , Cinética , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/sangue , Camundongos , Neuropeptídeo Y/administração & dosagemRESUMO
The present study was designed to determine if changes in dietary protein level and source are related to changes in VLDL lipid concentrations and VLDL binding by hepatic membranes and isolated hepatocytes. Male Wistar rats were fed cholesterol-free diets containing 10, 20 or 30 g/100 g casein or highly purified soybean protein for 4 wk. Hepatic, plasma and VLDL lipids, VLDL apo B-100 and VLDL uptake by isolated hepatocytes and VLDL binding to hepatic membrane were determined. Increasing casein or soybean protein level (from 10 to 30 g/100 g) in the diet increased VLDL apo B-100, indicating an increase in the number of VLDL particles. VLDL uptake by isolated hepatocytes and VLDL binding to hepatic membrane increased when the protein level increased from 10 to 20 g/100 g in the diet and decreased with 30 g/100 g protein, regardless of protein type. The dietary protein source did not affect plasma total cholesterol concentrations at any protein level. Feeding 20 g/100 g soybean protein compared with casein lowered plasma triglyceride concentrations and VLDL number as measured by decreased VLDL-protein, -phospholipid, -triglyceride, -cholesterol and -apo B-100. VLDL uptake by isolated hepatocytes and VLDL binding to hepatic membrane were higher in rats fed soybean protein than those fed casein. The higher VLDL uptake could be responsible for the hypotriglyceridemia in rats fed soybean protein.
Assuntos
Caseínas/administração & dosagem , Lipoproteínas VLDL/metabolismo , Proteínas de Soja/administração & dosagem , Animais , Apolipoproteína B-100 , Apolipoproteínas B/antagonistas & inibidores , Apolipoproteínas B/sangue , Relação Dose-Resposta a Droga , Hepatócitos/metabolismo , Metabolismo dos Lipídeos , Lipoproteínas VLDL/antagonistas & inibidores , Fígado/metabolismo , Masculino , Membranas/metabolismo , Concentração Osmolar , Ratos , Ratos WistarRESUMO
A newly synthesized benzoic acid derivative, (+)-(S)-p-[1-(p-tert-butylphenyl)-2-oxo-4-pyrrolidinyl]methoxybenzoic acid (S-2E), has the capacity to inhibit the biosynthesis of both sterol and fatty acids. Here, we report the mechanism by which S-2E lowers blood cholesterol and triglyceride levels. In the liver, S-2E was converted into its active metabolite, S-2E-CoA. S-2E-CoA noncompetitively inhibited the enzymatic activities of both 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase and acetyl-CoA carboxylase at K(i)=18.11 microM and K(i)=69.2 microM, respectively. Interestingly, pharmacokinetic experiments in rats showed that the concentration of S-2E-CoA in the liver was sufficient to inhibit the activities of HMG-CoA reductase and acetyl-CoA carboxylase, for example, when orally given to rats at 10 mg/kg. Indeed, S-2E (3-30 mg/kg) given orally suppressed the secretion rate of very-low-density lipoprotein (VLDL)-cholesterol and triglyceride in Triton WR-1339-injected rats. Furthermore, S-2E lowered the blood total cholesterol and triglyceride levels simultaneously in Zucker fatty rats. Collectively, S-2E may be useful in the treatment of familial hypercholesterolemia and mixed hyperlipidemia.
Assuntos
Benzoatos/síntese química , Benzoatos/metabolismo , Benzoatos/farmacologia , Hipolipemiantes/farmacologia , Pirrolidinonas/farmacologia , Acetil-CoA Carboxilase/antagonistas & inibidores , Acetil-CoA Carboxilase/metabolismo , Acil Coenzima A/antagonistas & inibidores , Acil Coenzima A/metabolismo , Administração Oral , Animais , Benzoatos/administração & dosagem , Benzoatos/farmacocinética , Colesterol/sangue , Colesterol/metabolismo , Ácidos Graxos/antagonistas & inibidores , Ácidos Graxos/biossíntese , Éteres de Hidroxibenzoatos , Hiperlipidemias/sangue , Hiperlipidemias/tratamento farmacológico , Hipolipemiantes/administração & dosagem , Hipolipemiantes/farmacocinética , Lipoproteínas VLDL/antagonistas & inibidores , Lipoproteínas VLDL/biossíntese , Lipoproteínas VLDL/química , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Obesidade/genética , Obesidade/metabolismo , Pirrolidinonas/administração & dosagem , Pirrolidinonas/farmacocinética , Ratos , Ratos Sprague-Dawley , Ratos Zucker/genética , Ratos Zucker/metabolismo , Esteróis/antagonistas & inibidores , Esteróis/biossíntese , Suspensões , Triglicerídeos/sangue , Triglicerídeos/metabolismoRESUMO
Recently, we generated mice lacking microsomal triglyceride transfer protein (MTP) in the liver (Mttp(Delta/Delta)) and demonstrated that very low density lipoprotein secretion from hepatocytes was almost completely blocked. The blockade in lipoprotein production was accompanied by mild to moderate hepatic steatosis, but the mice appeared healthy. Although hepatic MTP deficiency appeared to be innocuous, we hypothesized that a blockade in very low density lipoprotein secretion and the accompanying steatosis might increase the sensitivity of Mttp(Delta/Delta) livers to additional hepatic insults. To address this issue, we compared the susceptibility of Mttp(Delta/Delta) mice and Mttp(flox/flox) controls to hepatic injury from Escherichia coli lipopolysaccharides, concanavalin A, and Pseudomonas aeruginosa exotoxin A. At baseline, neither the Mttp(Delta/Delta) nor the Mttp(flox/flox) mice had elevated serum transaminases or histologic evidence of hepatic inflammation. After the administration of the toxins, however, the Mttp(Delta/Delta) mice manifested higher levels of transaminases and, unlike the Mttp(flox/flox) mice, developed histologic evidence of hepatic inflammation. The toxic challenge induced tumor necrosis factor-alpha to a similar extent in Mttp(Delta/Delta) and Mttp(flox/flox) mice, but other parameters of injury (e.g. chemokine transcript levels and lipid peroxides) were disproportionately increased in the Mttp(Delta/Delta) mice. Our results suggest that blocking lipoprotein secretion in the liver may increase the susceptibility of the liver to certain toxic challenges.