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1.
Clin Exp Pharmacol Physiol ; 49(8): 805-812, 2022 08.
Article in English | MEDLINE | ID: mdl-35577580

ABSTRACT

Atherosclerosis is associated with a haemostatic imbalance characterized by excessive activation of pro-inflammatory and pro-coagulant pathways. Non-vitamin K antagonists oral anticoagulant (NOACs) may reduce the incidence of cardiovascular events, cerebral ischemia, thromboembolic events and atherosclerosis. Chronic inflammation, vascular proliferation and the development of atherosclerosis is also influenced by 25-hydroxycholesterol (25-OHC). The aim of the study was to assess the effect of rivaroxaban and dabigatran on the messenger RNA (mRNA) expression of anti-inflammatory cytokines transforming growth factor ß (TGF-ß), interleukin (IL)-37, IL-35 as well as of pro-inflammatory cytokines IL-18 and IL-23, in endothelial cells damaged by 25-OHC. Human umbilical vascular endothelial cells (HUVECs) were treated with 25-OHC (10 µg/mL), rivaroxaban (100, 500 ng/mL), dabigatran (100, 500 ng/mL), 25-OHC + rivaroxaban, and 25-OHC + dabigatran. The mRNA expression of TGF-ß, IL-37, IL-35 subunits EBI3 and p35, IL-18, and IL-23 was analysed using real-time polymerase chain reaction (PCR). The results showed that 25-OHC decreased TGF-ß and IL-37 mRNA expression and increased EBI3, p35, IL-18, IL-23 mRNA expression in endothelial cell as compared to an untreated control (P < .05). Messenger RNA expression of TGF-ß and IL-37 significantly increased following stimulation with rivaroxaban and dabigatran as compared to an untreated control (P < .01). In HUVECs pre-treated with oxysterol, rivaroxaban and dabigatran increased mRNA expression of TGF-ß, IL-37 and decreased mRNA expression of EBI3, p35, IL-23 and IL-18 as compared to 25-OHC (P < .01). Our finding suggests that both rivaroxaban and dabigatran inhibit the inflammatory activation caused by oxysterol in vitro.


Subject(s)
Atherosclerosis , Cytokines , Dabigatran , Human Umbilical Vein Endothelial Cells , Hydroxycholesterols , Rivaroxaban , Administration, Oral , Anticoagulants/pharmacology , Anticoagulants/therapeutic use , Atherosclerosis/drug therapy , Atherosclerosis/genetics , Atherosclerosis/immunology , Atrial Fibrillation/drug therapy , Cytokines/genetics , Cytokines/immunology , Dabigatran/pharmacology , Dabigatran/therapeutic use , Endothelial Cells/drug effects , Endothelial Cells/immunology , Human Umbilical Vein Endothelial Cells/drug effects , Human Umbilical Vein Endothelial Cells/immunology , Humans , Hydroxycholesterols/administration & dosage , Hydroxycholesterols/adverse effects , Hydroxycholesterols/pharmacology , Interleukin-18/genetics , Interleukin-18/immunology , Interleukin-23/genetics , Interleukin-23/immunology , Oxysterols/administration & dosage , Oxysterols/adverse effects , Oxysterols/pharmacology , RNA, Messenger/genetics , RNA, Messenger/immunology , Rivaroxaban/pharmacology , Rivaroxaban/therapeutic use , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta/immunology
2.
Aging (Albany NY) ; 13(16): 20149-20163, 2021 08 18.
Article in English | MEDLINE | ID: mdl-34406977

ABSTRACT

Middle cerebral artery occlusion (MCAO) injury refers to impaired blood supply to the brain that is caused by a cerebrovascular disease, resulting in local brain tissue ischemia, hypoxic necrosis, and rapid neurological impairment. Nevertheless, the mechanisms involved are unclear, and pharmacological interventions are lacking. 25-Hydroxycholesterol (25-HC) was reported to be involved in cholesterol and lipid metabolism as an oxysterol molecule. This study aimed to determine whether 25-HC exerts a cerebral protective effect on MCAO injury and investigate its potential mechanism. 25-HC was administered prior to reperfusion in a mouse model of MCAO injury. 25-HC evidently decreased infarct size induced by MCAO and enhanced brain function. It reduced stimulator of interferon gene (STING) activity and regulated mTOR to inhibit autophagy and induce cerebral ischemia tolerance. Thus, 25-HC improved MCAO injury through the STING channel. As indicated in this preliminary study, 25-HC improved MCAO injury by inhibiting STING activity and autophagy as well as by reducing brain nerve cell apoptosis. Thus, it is a potential treatment drug for brain injury.


Subject(s)
Hydroxycholesterols/administration & dosage , Infarction, Middle Cerebral Artery/drug therapy , Membrane Proteins/metabolism , Protective Agents/administration & dosage , Reperfusion Injury/drug therapy , Animals , Autophagy/drug effects , Humans , Infarction, Middle Cerebral Artery/genetics , Infarction, Middle Cerebral Artery/metabolism , Infarction, Middle Cerebral Artery/physiopathology , Male , Membrane Proteins/genetics , Mice , Mice, Inbred C57BL , Reperfusion Injury/genetics , Reperfusion Injury/metabolism , Reperfusion Injury/physiopathology , TOR Serine-Threonine Kinases/genetics , TOR Serine-Threonine Kinases/metabolism
3.
Int Immunopharmacol ; 96: 107643, 2021 Jul.
Article in English | MEDLINE | ID: mdl-33878616

ABSTRACT

Hepatic ischemia reperfusion (I/R) injury remains a major obstacle in liver transplantation, however an effective treatment to mitigate this injury is lacking. 25-Hydroxycholesterol (25HC) is a kind of oxysterol involved in inflammatory and immune responses. However, its function and the underlying mechanism on rat hepatic I/R injury has not been explored. A well-established rat model of partial warm ischemia reperfusion injury was performed. 25HC was intraperitoneally administrated 4 h before ischemia. The results verified that 25HC pretreatment effectively mitigated liver I/R injury, which was demonstrated by lower serum levels of transaminases, histology injury score and less apoptosis. Mechanistically, 25HC pretreatment activated PINK1/Parkin dependent mitophagy and inhibited the NLRP3 inflammasome. Via using mitophagy inhibitor 3-methyladenine (3-MA), we further found that 3-MA counteracted the protective effect of 25HC on hepatic I/R injury and the NLRP3 inflammasome. In conclusion, 25HC pretreatment ameliorates rat hepatic I/R injury, and this protective effect may be dependent on activating mitophagy and inhibiting NLRP3 inflammasome activation.


Subject(s)
Hydroxycholesterols/pharmacology , Liver Diseases/prevention & control , Mitophagy/drug effects , Protective Agents/pharmacology , Reperfusion Injury/prevention & control , Adenine/analogs & derivatives , Adenine/pharmacology , Adenine/therapeutic use , Alanine Transaminase/blood , Animals , Apoptosis/drug effects , Aspartate Aminotransferases/blood , Disease Models, Animal , Hydroxycholesterols/administration & dosage , Hydroxycholesterols/metabolism , Liver Diseases/etiology , Liver Diseases/metabolism , Liver Diseases/pathology , Male , NLR Family, Pyrin Domain-Containing 3 Protein/antagonists & inhibitors , Protective Agents/administration & dosage , Protein Kinases/metabolism , Rats, Sprague-Dawley , Reperfusion Injury/metabolism , Reperfusion Injury/pathology , Ubiquitin-Protein Ligases/metabolism
4.
J Cell Mol Med ; 22(11): 5494-5503, 2018 11.
Article in English | MEDLINE | ID: mdl-30091835

ABSTRACT

Acute lung injury (ALI) is mainly caused by uncontrolled inflammatory response, and it remains without effective therapeutic options. 25-hydroxycholesterol (25HC) has been reported to be a potent regulator of inflammation. The aim of this study was to investigate the effects of 25HC on lipopolysaccharide (LPS)-induced ALI. C57BL/6 mice were pretreated with 25HC intraperitoneally before intratracheal exposure to LPS. Our results showed that 25HC pretreatment improved survival rate, attenuated the pathological changes of the lung and decreased the release of inflammatory cytokines in mice. Consistently, 25HC reduced expression of Toll-like receptor (TLR4)-mediated inflammatory cytokines in vitro. These effects of 25HC were obtained by preventing LPS binding to TLR4 via interaction with myeloid differentiation protein 2 (MD-2). Crystal structure analysis suggested that 25HC could bind MD-2 with high affinity into its hydrophobic pocket. Furthermore, LPS-induced activation of Akt/NF-κB pathway was partially down-regulated by 25HC pretreatment. In summary, this study demonstrates that 25HC could inhibit the overwhelming inflammatory response through MD-2 interaction, which suppresses Akt/NF-κB signalling pathway. These findings suggest 25HC may be a promising candidate for ALI prevention.


Subject(s)
Acute Lung Injury/drug therapy , Hydroxycholesterols/administration & dosage , Lymphocyte Antigen 96/genetics , Toll-Like Receptor 4/genetics , Acute Lung Injury/chemically induced , Acute Lung Injury/genetics , Acute Lung Injury/pathology , Animals , Anti-Inflammatory Agents/administration & dosage , Bronchoalveolar Lavage Fluid , Disease Models, Animal , Humans , Inflammation/chemically induced , Inflammation/drug therapy , Inflammation/genetics , Inflammation/pathology , Lipopolysaccharides/toxicity , Lung/pathology , Mice , NF-kappa B/genetics , Protective Agents , RAW 264.7 Cells , Signal Transduction/drug effects
5.
Redox Biol ; 17: 423-431, 2018 07.
Article in English | MEDLINE | ID: mdl-29883958

ABSTRACT

It is now established that cholesterol oxidation products (oxysterols) are involved in several events underlying Alzheimer's disease (AD) pathogenesis. Of note, certain oxysterols cause neuron dysfunction and degeneration but, recently, some of them have been shown also to have neuroprotective effects. The present study, which aimed to understand the potential effects of 24-hydroxycholesterol (24-OH) against the intraneuronal accumulation of hyperphosphorylated tau protein, stressed these latter effects. A beneficial effect of 24-OH was demonstrated in SK-N-BE neuroblastoma cells, and is due to its ability to modulate the deacetylase sirtuin 1 (SIRT1), which contributes to preventing the neurotoxic accumulation of the hyperphosphorylated tau protein. Unlike 24-OH, 7-ketocholesterol (7-K) did not modulate the SIRT1-dependent neuroprotective pathway. To confirm the neuroprotective role of 24-OH, in vivo experiments were run on mice that express human tau without spontaneously developing tau pathology (hTau mice), by means of the intracerebroventricular injection of 24-OH. 24-OH, unlike 7-K, was found to completely prevent the hyperphosphorylation of tau induced by amyloid ß monomers. These data highlight the importance of preventing the loss of 24-OH in the brain, and of maintaining high levels of the enzyme SIRT1, in order to counteract neurodegeneration.


Subject(s)
Alzheimer Disease/genetics , Hydroxycholesterols/metabolism , Sirtuin 1/genetics , tau Proteins/genetics , Alzheimer Disease/metabolism , Alzheimer Disease/pathology , Alzheimer Disease/prevention & control , Amyloid beta-Peptides/genetics , Amyloid beta-Peptides/metabolism , Animals , Brain/metabolism , Brain/pathology , Disease Models, Animal , Humans , Hydroxycholesterols/administration & dosage , Ketocholesterols/administration & dosage , Mice , Mice, Transgenic , Neurons/drug effects , Neurons/metabolism , Neurons/pathology , Neuroprotective Agents/metabolism , Oxidation-Reduction , tau Proteins/metabolism
6.
Toxicol In Vitro ; 45(Pt 1): 10-18, 2017 Dec.
Article in English | MEDLINE | ID: mdl-28739487

ABSTRACT

The oxysterol 27-hydroxycholesterol (27HC) is a selective estrogen receptor modulator (SERMs), which like endogenous estrogen 17ß-estradiol (E2) induces the proliferation of ER-positive breast cancer cells in vitro. Interestingly, the observation that 27HC induces adverse effects in neural system, distinguishing it from E2. It has been suggested that high levels of circulating cholesterol increase the entry of 27HC into the brain, which may induce learning and memory impairment. Based on this evidence, 27HC may be associated with neurodegenerative processes and interrupted cholesterol homeostasis in the brain. However, the biological events that participate in this process remain largely elusive. In the present study, we demonstrated that 27HC induced apparent cellular senescence in nerve cells. Senescence-associated ß-galactosidase (SA-ß-Gal) assay revealed that 27HC induced senescence in both BV2 cells and PC12 cells. Furthermore, we demonstrated that 27HC promoted the accumulation of cellular reactive oxygen species (ROS) in nerve cells and subsequently activation of IL-6/STAT3 signaling pathway. Notably, treatment with the ROS scavenger N-acetylcysteine (NAC) markedly blocked 27HC-induced ROS production and activation of IL-6/STAT3 signaling pathway. Either blocking the generation of ROS or inhibition of IL-6/STAT3 both attenuated 27HC-induced cellular senescence. In sum, these findings not only suggested a mechanism whereby 27HC induced cellular senescence in nerve cells, but also helped to recognize the 27HC as a novel harmful factor in neurodegenerative diseases.


Subject(s)
Cell Survival/drug effects , Hydroxycholesterols/toxicity , Interleukin-6/metabolism , Microglia , Reactive Oxygen Species/metabolism , STAT3 Transcription Factor/metabolism , Animals , Cell Line , Cell Survival/physiology , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation/drug effects , Humans , Hydroxycholesterols/administration & dosage , Mice , Pheochromocytoma , Rats
7.
PLoS One ; 12(3): e0173749, 2017.
Article in English | MEDLINE | ID: mdl-28323848

ABSTRACT

We investigated effects of 7-oxygenated cholesterol derivatives present in atherosclerotic lesions, 7α-hydroxycholesterol (7αOHChol), 7ß-hydroxycholesterol (7ßOHChol), and 7-ketocholesterol (7K), on IL-8 expression. Transcript levels of IL-8 and secretion of its corresponding gene product by monocytes/macrophages were enhanced by treatment with 7αOHChol and, to a lesser extent, 7K, but not by 7ßOHChol. The 7-oxygenated cholesterol derivatives, however, did not change transcription of the IL-8 gene in vascular smooth muscle cells. 7αOHChol-induced IL-8 gene transcription was inhibited by cycloheximide and Akt1 downregulation, but not by OxPAPC. Expression of C5a receptor was upregulated after stimulation with 7αOHChol, but not with 7K and 7ßOHChol, and a specific antagonist of C5a receptor inhibited 7αOHChol-induced IL-8 gene expression in a dose dependent manner. Pharmacological inhibitors of PI3K and MEK almost completely inhibited expression of both IL-8 and cell-surface C5a receptor induced by 7αOHChol. These results indicate that 7-oxygenated cholesterol derivatives have differential effects on monocyte/macrophage expression of IL-8 and C5a receptor and that C5a receptor is involved in 7αOHChol-induced IL-8 expression via PI3K and MEK.


Subject(s)
Hydroxycholesterols/metabolism , Interleukin-8/metabolism , Macrophages/metabolism , Monocytes/metabolism , Receptor, Anaphylatoxin C5a/metabolism , Aniline Compounds/pharmacology , Butadienes/pharmacology , Cell Membrane/drug effects , Cell Membrane/metabolism , Cells, Cultured , Chromones/pharmacology , Dose-Response Relationship, Drug , Enzyme Inhibitors/pharmacology , Gene Expression/drug effects , Gene Expression/physiology , Humans , Hydroxycholesterols/administration & dosage , Ketocholesterols/administration & dosage , Ketocholesterols/metabolism , MAP Kinase Kinase Kinases/antagonists & inhibitors , MAP Kinase Kinase Kinases/metabolism , Macrophages/drug effects , Monocytes/drug effects , Morpholines/pharmacology , Nitriles/pharmacology , Phosphatidylinositol 3-Kinases/metabolism , Phosphoinositide-3 Kinase Inhibitors , Receptor, Anaphylatoxin C5a/antagonists & inhibitors , Tetrahydronaphthalenes/pharmacology , Transcription, Genetic/drug effects , Transcription, Genetic/physiology
8.
Clin Pharmacol Drug Dev ; 6(1): 19-26, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27138546

ABSTRACT

The objectives of this analysis were to establish the exposure-response relationship between plasma rifampicin and 4ß-hydroxycholesterol (4ßHC) concentration and to estimate the effect of weak, moderate, and potent CYP3A induction. Plasma rifampicin and 4ßHC concentration-time data from a drug-drug interaction study with rifampicin 600 mg were used for model development. An indirect response model with an effect compartment described the relationship between rifampicin and 4ßHC concentrations. The model predicted that the equilibration t1/2 and 4ßHC t1/2 were 72.8 and 142 hours, respectively. EM50 and Emax of rifampicin induction were 32.6 µg and 8.39-fold, respectively. The population PK-PD model was then used to simulate the effects of rifampicin 10, 20, and 100 mg on plasma 4ßHC for up to 21 days, in which rifampicin 10, 20, and 100 mg were used to represent weak, moderate, and strong inducers, respectively. The model-predicted median (5th, 95th percentiles) 1.13 (1.04, 1.44)-, 1.28 (1.10, 1.71)-, and 2.10 (1.45, 3.49)-fold increases in plasma 4ßHC after 14-day treatment with rifampicin 10, 20, and 100 mg, respectively. A new drug candidate can likely be classified as a weak, moderate, or strong inducer if baseline-normalized plasma 4ßHC increases by <1.13-, 1.13- to 2.10-, or >2.10-fold, respectively, after 14 days of dosing.


Subject(s)
Cytochrome P-450 CYP3A Inducers/administration & dosage , Hydroxycholesterols/blood , Rifampin/administration & dosage , Adult , Cytochrome P-450 CYP3A Inducers/blood , Cytochrome P-450 CYP3A Inducers/pharmacokinetics , Drug Interactions , Female , Healthy Volunteers , Humans , Hydroxycholesterols/administration & dosage , Hydroxycholesterols/pharmacology , Male , Middle Aged , Models, Theoretical , Rifampin/blood , Rifampin/pharmacokinetics , Young Adult
9.
J Lipid Res ; 57(11): 2005-2014, 2016 11.
Article in English | MEDLINE | ID: mdl-27647838

ABSTRACT

The 24(S)-hydroxycholesterol (24S-OHC), which plays an important role in maintaining brain cholesterol homeostasis, has been shown to possess neurotoxicity. We have previously reported that 24S-OHC esterification by ACAT1 and the resulting lipid droplet (LD) formation are responsible for 24S-OHC-induced cell death. In the present study, we investigate the functional roles of 24S-OHC esters and LD formation in 24S-OHC-induced cell death, and we identify four long-chain unsaturated fatty acids (oleic acid, linoleic acid, arachidonic acid, and DHA) with which 24S-OHC is esterified in human neuroblastoma SH-SY5Y cells treated with 24S-OHC. Here, we find that cotreatment of cells with 24S-OHC and each of these four unsaturated fatty acids increases prevalence of the corresponding 24S-OHC ester and exacerbates induction of cell death as compared with cell death induced by treatment with 24S-OHC alone. Using electron microscopy, we find in the present study that 24S-OHC induces formation of LD-like structures coupled with enlarged endoplasmic reticulum (ER) lumina, and that these effects are suppressed by treatment with ACAT inhibitor. Collectively, these results illustrate that ACAT1-catalyzed esterification of 24S-OHC with long-chain unsaturated fatty acid followed by formation of atypical LD-like structures at the ER membrane is a critical requirement for 24S-OHC-induced cell death.


Subject(s)
Acetyl-CoA C-Acetyltransferase/genetics , Brain/metabolism , Hydroxycholesterols/administration & dosage , Lipid Droplets/metabolism , Neurons/metabolism , Arachidonic Acid/administration & dosage , Arachidonic Acid/metabolism , Brain/drug effects , Brain/pathology , Cell Death/drug effects , Cell Line, Tumor , Endoplasmic Reticulum/drug effects , Endoplasmic Reticulum/metabolism , Esterification/drug effects , Gene Expression Regulation/drug effects , Humans , Hydroxycholesterols/metabolism , Linoleic Acid/administration & dosage , Linoleic Acid/metabolism , Lipid Droplets/chemistry , Lipid Droplets/drug effects , Neuroblastoma/metabolism , Neurons/pathology , Oleic Acid/administration & dosage , Oleic Acid/metabolism
11.
Biofactors ; 42(1): 80-92, 2016.
Article in English | MEDLINE | ID: mdl-26669587

ABSTRACT

Macrophages play a crucial role in atherosclerosis progression. Classically activated M1 macrophages have been found in rupture-prone atherosclerotic plaques whereas alternatively activated macrophages, M2, localize in stable plaque. Macrophage accumulation of cholesterol and of its oxidized derivatives (oxysterols) leads to the formation of foam cells, a hallmark of atherosclerotic lesions. In this study, the effects of oxysterols in determining the functional polarization of human macrophages were investigated. Monocytes, purified from peripheral blood mononuclear cells of healthy donors, were differentiated into macrophages (M0) and treated with an oxysterol mixture, cholesterol, or ethanol, every 4 H for a total of 4, 8, and 12 H. The administration of the compounds was repeated in order to maintain the levels of oxysterols constant throughout the treatment. Compared with ethanol treatment, the oxysterol mixture decreased the surface expression of CD36 and CD204 scavenger receptors and reduced the amount of reactive oxygen species whereas it did not affect either cell viability or matrix metalloprotease-9 activity. Moreover, the oxysterol mixture increased the expression of both liver X receptor α and ATP-binding cassette transporter 1. An enhanced secretion of the immunoregulatory cytokine IL-10 accompanied these events. The results supported the hypothesis that the constant levels of oxysterols and, in particular, of 27-hydroxycholesterol stimulate macrophage polarization toward the M2 immunomodulatory functional phenotype, contributing to the stabilization of atherosclerotic plaques.


Subject(s)
Atherosclerosis/drug therapy , Hydroxycholesterols/administration & dosage , Macrophages/drug effects , Plaque, Atherosclerotic/drug therapy , ATP-Binding Cassette Transporters/biosynthesis , Atherosclerosis/metabolism , Atherosclerosis/pathology , CD36 Antigens/metabolism , Cell Polarity/drug effects , Cell Survival/drug effects , Cholesterol/administration & dosage , Cholesterol/metabolism , Ethanol/administration & dosage , Humans , Interleukin-10/metabolism , Liver X Receptors , Macrophages/metabolism , Matrix Metalloproteinase 9/biosynthesis , Monocytes/drug effects , Orphan Nuclear Receptors/biosynthesis , Plaque, Atherosclerotic/metabolism , Plaque, Atherosclerotic/pathology , Reactive Oxygen Species/metabolism , Scavenger Receptors, Class A/metabolism
12.
Biochem J ; 471(3): 369-79, 2015 Nov 01.
Article in English | MEDLINE | ID: mdl-26310456

ABSTRACT

Pcyt2 (CTP:phosphoethanolamine cytidylyltransferase) is the rate-limiting enzyme in mammalian PE (phosphatidylethanolamine) biosynthesis. Previously, we reported that Pcyt2 mRNA levels increased in several types of cells after serum starvation, an effect that could be suppressed by supplementation with low-density lipoprotein or 25-HC (25-hydroxycholesterol). Transcription of Hmgcr, which encodes 3-hydroxy-3-methylglutaryl-CoA reductase, is also suppressed by 25-HC in the same dose-dependent manner. Nevertheless, a sterol-regulatory element was not detected in the Pcyt2 promoter region. The important element for transcriptional control of Pcyt2 by 25-HC (1.25 µM) was determined to reside between -56 and -36 on the basis of analysis with several Pcyt2 promoter deletion-luciferase reporters in NIH 3T3 cells. Using the yeast one-hybrid system, we found that NF-Y (nuclear factor-Y) binds at C(-37)CAAT(-41) and YY1 (Yin Yang1) binds at C(-42)AT(-40) in the Pcyt2 promoter. Endogenous NF-Y and YY1 bind clearly and competitively to these sites and are important for basal Pcyt2 transcription. Moreover, NF-Y binds to the Hmgcr promoter at C(-14)CA(-12) in gel-shift analysis, and suppression of the basal luciferase activity of the Hmgcr promoter-reporter construct (-30/+61) by 25-HC was abolished when C(-14)CA(-12) was mutated. Furthermore, transcriptional suppression of Pcyt2 by 25-HC was reduced following knockdown targeting of NF-YA or YY1. ChIP analysis revealed that 25-HC inhibited the interaction between NF-Y and RNA polymerase II on the Pcyt2 and Hmgcr promoters. On the basis of these results, we conclude that NF-Y and YY1 are important for the basal transcription of Pcyt2 and that NF-Y is involved in the inhibitory effects of 25-HC on Pcyt2 transcription.


Subject(s)
CCAAT-Binding Factor/metabolism , RNA Nucleotidyltransferases/genetics , Transcription, Genetic/drug effects , YY1 Transcription Factor/metabolism , Animals , CCAAT-Binding Factor/genetics , DNA-Binding Proteins/genetics , Humans , Hydroxycholesterols/administration & dosage , Hydroxycholesterols/metabolism , Hydroxymethylglutaryl CoA Reductases/biosynthesis , Mice , NIH 3T3 Cells , Promoter Regions, Genetic , YY1 Transcription Factor/genetics
14.
Drug Res (Stuttg) ; 65(5): 231-7, 2015 May.
Article in English | MEDLINE | ID: mdl-24668576

ABSTRACT

BACKGROUND: Recent studies have shown that hypercholesterolemia, besides being a risk factor for cardiovascular diseases, has also toxic effects on central nervous system. The design of the present study was to investigate the effects of dietary cholesterol and oxidized cholesterol on cognitive function. METHODS: Male Wistar rats were randomly divided into 3 groups. The animals were fed with three normal, 2% cholesterol-rich, and 2% oxidized cholesterol-rich diets for 14 weeks. Memory impairment was analyzed by passive avoidance test. Coenzyme Q10 content was also measured by a validate RP-HPLC method. Besides, lipid peroxidation in serum and brain tissue was determined by malondialdehyde concentration measurement. RESULTS: The results showed that feeding rats with high oxidized cholesterol diet for 14 weeks significantly impaired the cognitive function compared to the normal (P<0.001) and high cholesterol-fed groups (P<0.01). The memory impairment was positively correlated to the serum level of the oxidized LDL; it was significantly associated with the increased malondialdehyde concentration on the brain tissue of both groups (P<0.05 and P<0.001, respectively). The total antioxidant level in the serum was also decreased in rats fed with the oxidized cholesterol (P<0.05). Moreover, the brain coenzyme Q10 content was significantly declined in the animals fed with the oxidized cholesterol-rich diet compared to the animals fed with the normal (P<0.01) and cholesterol-rich diets (P<0.05). CONCLUSION: The results suggested that the high dietary intake of the oxidized-cholesterol might impair the memory that could be correlated to the oxidative stress and declined the coenzyme Q10 content of the brain tissue.


Subject(s)
Avoidance Learning/drug effects , Cholesterol, Dietary/adverse effects , Hydroxycholesterols/adverse effects , Memory Disorders/chemically induced , Animals , Antioxidants/metabolism , Brain/drug effects , Brain/metabolism , Hydroxycholesterols/administration & dosage , Lipid Peroxidation/drug effects , Lipoproteins, LDL/blood , Male , Malondialdehyde/blood , Memory Disorders/blood , Memory Disorders/psychology , Oxidation-Reduction , Rats , Ubiquinone/analogs & derivatives , Ubiquinone/metabolism
15.
Int J Med Sci ; 10(12): 1615-24, 2013.
Article in English | MEDLINE | ID: mdl-24151433

ABSTRACT

BACKGROUND: The activation of the renin-angiotensin system (RAS) and lipid disorders are major risk factors in progressive chronic kidney disease. This study aimed to investigate the potential synergistic mechanisms of RAS activation and lipid disorders that contribute to glomerulosclerosis. MATERIALS AND METHODS: Human renal mesangial cells (HMCs) were treated with 10(-7) mol/L angiotensin II (Ang II) or with 30 µg/ml cholesterol and 1 µg/ml 25-hydroxycholesterol (lipid loading) for 24 hours. Lipid accumulation in the cells was evaluated by Oil Red O staining and intracellular cholesterol quantitative assays. The gene and protein expression of molecules in the low-density lipoprotein receptor (LDLr) pathway, the RAS family, and the extracellular matrix were examined by real-time polymerase chain reaction and Western blotting. The translocation of sterol regulatory element-binding protein (SREBP) cleavage activating protein (SCAP), which escorts SREBP-2 from the endoplasmic reticulum (ER) to the Golgi, was examined by immunofluorescent staining. RESULTS: Ang II increased lipid droplet accumulation in HMCs. Further analysis revealed that Ang II increased the mRNA and protein expression of LDLr, SCAP, and SREBP-2. This increase was correlated with an enhanced translocation of the SCAP/SREBP-2 complex from the ER to the Golgi in HMCs that was induced by Ang II, thereby activating LDLr gene transcription. Interestingly, lipid loading increased the mRNA and protein expression of angiotensinogen, Ang II, renin, angiotensin-converting enzyme, angiotensin II type 1 receptor, and type 2 receptor in HMCs with increased mRNA and protein expression of collagen I, α-smooth muscle actin, and fibronectin. CONCLUSIONS: This study demonstrates that the interaction of RAS activation and lipid disorders accelerates the progression of glomerulosclerosis.


Subject(s)
Angiotensin II/administration & dosage , Diabetic Nephropathies/genetics , Lipid Metabolism/drug effects , ras Proteins/genetics , Angiotensin II/metabolism , Cholesterol/administration & dosage , Diabetic Nephropathies/metabolism , Diabetic Nephropathies/pathology , Disease Progression , Endoplasmic Reticulum/metabolism , Gene Expression Regulation/drug effects , Humans , Hydroxycholesterols/administration & dosage , Intracellular Signaling Peptides and Proteins/biosynthesis , Membrane Proteins/biosynthesis , Mesangial Cells/drug effects , Mesangial Cells/metabolism , Receptor, Angiotensin, Type 1/metabolism , Receptors, Lipoprotein/biosynthesis , Receptors, Lipoprotein/metabolism , Renin-Angiotensin System/genetics , Sterol Regulatory Element Binding Protein 2/biosynthesis , Sterol Regulatory Element Binding Proteins/biosynthesis , ras Proteins/metabolism
16.
Lipids ; 47(5): 483-93, 2012 May.
Article in English | MEDLINE | ID: mdl-22426832

ABSTRACT

This study explores the pharmacokinetics of 22-S-hydroxycholesterol (22SHC) in vivo in rats. We also carried out a metabolic study to explore whether the beneficial effects observed of 22SHC on glucose and lipid metabolism in vitro could be seen in vivo in rats. In the pharmacokinetic study, rats were given 50 mg/kg of [³H]22-S-hydroxycholesterol before absorption, distribution and excretion were monitored. In the metabolic study, the effect of 22SHC (30 mg/kg/day for 3 weeks) in rats on body weight gain [chow and high-fat diet (HFD)], serum lipids triacylglycerol (TAG) content and gene expression in liver and skeletal muscle were examined. Results showed that 22SHC was well absorbed after oral administration and distributed to most organs and mainly excreted in feces. Rats receiving 22SHC gained less body weight than their controls regardless whether the animals received chow diet or HFD. Moreover, we observed that animals receiving HFD had elevated levels of serum TAG while this was not observed for animals on HFD supplemented with 22SHC. The amount of TAG in liver was reduced after 22SHC treatment in animals receiving either chow diet or HFD. Gene expression analysis revealed that two genes (carnitine palmitoyltransferase 2 and uncoupling protein 3) involved in fatty acid oxidation and energy dissipation were increased in liver. Ucp3 expression (both protein and mRNA level) was increased in skeletal muscle, but insulin-stimulated glucose uptake and TAG content were unchanged. In conclusion, 22SHC seems to be an interesting model substance in the search of treatments for disorders involving aberrations in lipid metabolism.


Subject(s)
Dietary Supplements , Hydroxycholesterols/administration & dosage , Hydroxycholesterols/pharmacology , Liver/drug effects , Liver/metabolism , Triglycerides/metabolism , Weight Gain/drug effects , Administration, Oral , Animals , Fatty Acids/metabolism , Male , Rats , Rats, Sprague-Dawley , Triglycerides/analysis
17.
Inflammation ; 35(4): 1302-7, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22391743

ABSTRACT

Oxysterols are structurally similar to cholesterol, but are characterized by one or more additional oxygen-containing functional groups. These compounds are implicated in inflammation given their ability to cause irreversible damage to vascular cells. The aim of this study was to study the alteration of some inflammatory biomarkers in Wistar rats in response to dietary oxysterols. Eighteen rats were randomly divided into three groups of six rats each. A standard diet supplemented with 1% (w/w) pure cholesterol (Chol group) or 1% (w/w) of an oxidized cholesterol mixture (COPs group) was fed for 8 weeks. Blood serum was separated; abdominal, pericardial, and epididymal adipose tissue was removed carefully. The COPs subjects exhibited significant increase in blood pressure and serum triacylgycerols as well as increased body fat index and pericardic, abdominal, and epididymal adipose tissue. These effects were accompanied by elevated circulating levels of plasma high-sensitivity C-reactive protein, tumor necrosis factor alpha, and resistin. We suggest that dietary oxysterols have an important pro-inflammatory effect.


Subject(s)
Cholesterol, Dietary/analogs & derivatives , Cholesterol, Dietary/administration & dosage , Hydroxycholesterols/administration & dosage , Inflammation/chemically induced , Ketocholesterols/administration & dosage , Adipose Tissue/drug effects , Adipose Tissue/metabolism , Animals , Biomarkers/analysis , Blood Pressure/drug effects , C-Reactive Protein/analysis , Cholesterol, Dietary/blood , Random Allocation , Rats , Rats, Wistar , Resistin/blood , Triglycerides/blood , Tumor Necrosis Factor-alpha/blood
18.
Nat Med ; 13(10): 1185-92, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17873880

ABSTRACT

The cardioprotective effects of estrogen are mediated by receptors expressed in vascular cells. Here we show that 27-hydroxycholesterol (27HC), an abundant cholesterol metabolite that is elevated with hypercholesterolemia and found in atherosclerotic lesions, is a competitive antagonist of estrogen receptor action in the vasculature. 27HC inhibited both the transcription-mediated and the non-transcription-mediated estrogen-dependent production of nitric oxide by vascular cells, resulting in reduced estrogen-induced vasorelaxation of rat aorta. Furthermore, increasing 27HC levels in mice by diet-induced hypercholesterolemia, pharmacologic administration or genetic manipulation (by knocking out the gene encoding the catabolic enzyme CYP7B1) decreased estrogen-dependent expression of vascular nitric oxide synthase and repressed carotid artery reendothelialization. As well as antiestrogenic effects, there were proestrogenic actions of 27HC that were cell-type specific, indicating that 27HC functions as an endogenous selective estrogen receptor modulator (SERM). Taken together, these studies point to 27HC as a contributing factor in the loss of estrogen protection from vascular disease.


Subject(s)
Cardiotonic Agents/antagonists & inhibitors , Cardiotonic Agents/pharmacology , Estrogens/pharmacology , Hydroxycholesterols/pharmacology , Selective Estrogen Receptor Modulators/pharmacology , Animals , Aorta, Thoracic/drug effects , Binding, Competitive/drug effects , Cardiotonic Agents/metabolism , Cell Culture Techniques , Cell Line , Cells, Cultured , Cholesterol, Dietary/administration & dosage , DNA, Complementary , Dose-Response Relationship, Drug , Drug Administration Schedule , Estrogens/metabolism , Female , Glutathione Transferase/metabolism , Humans , Hydroxycholesterols/administration & dosage , Hydroxycholesterols/blood , Inhibitory Concentration 50 , Injections, Subcutaneous , Kidney/cytology , Kinetics , Male , Mice , Mice, Knockout , Nitric Oxide/antagonists & inhibitors , Nitric Oxide Synthase Type II/antagonists & inhibitors , Nitric Oxide Synthase Type III , RNA, Messenger/metabolism , Receptors, Estrogen/antagonists & inhibitors , Receptors, Estrogen/genetics , Recombinant Fusion Proteins/antagonists & inhibitors , Selective Estrogen Receptor Modulators/administration & dosage , Selective Estrogen Receptor Modulators/blood , Vasodilation/drug effects
19.
Free Radic Biol Med ; 43(5): 695-701, 2007 Sep 01.
Article in English | MEDLINE | ID: mdl-17664133

ABSTRACT

The oxysterols 7beta-hydroxycholesterol and 7-ketocholesterol are cholesterol autoxidation products. These two oxysterols are formed as a result of low density lipoprotein oxidation and in a study on biomarkers for oxidative stress in patients with atherosclerosis, 7beta-hydroxycholesterol was found to be the strongest predictor of progression of carotid atherosclerosis. Interconversion of 7beta-hydroxycholesterol and 7-ketocholesterol in vitro has been reported recently, using recombinant 11beta-hydroxysteroid dehydrogenase or rodent liver microsomes. In this study deuterium-labeled 7beta-hydroxycholesterol or 7-ketocholesterol was administered intravenously to two healthy volunteers and blood samples were collected at different time points. The mean half-life for elimination of 7beta-hydroxycholesterol from the circulation was estimated to be 1.9 h. The corresponding half-life for 7-ketocholesterol was estimated to be 1.5 h. Infusion of deuterium-labeled 7-ketocholesterol resulted in labeling of 7beta-hydroxycholesterol and vice versa. In addition, the biological within-day and between-day variations of the two oxysterols were determined. In summary, the present investigation clearly shows an interconversion of 7beta-hydroxycholesterol and 7-ketocholesterol in humans.


Subject(s)
Hydroxycholesterols/blood , Ketocholesterols/blood , Oxidative Stress , Adult , Biomarkers/blood , Female , Health , Humans , Hydroxycholesterols/administration & dosage , Hydroxycholesterols/pharmacology , Infusions, Intravenous , Ketocholesterols/administration & dosage , Ketocholesterols/pharmacology , Male , Middle Aged
20.
Biochem Biophys Res Commun ; 359(1): 46-50, 2007 Jul 20.
Article in English | MEDLINE | ID: mdl-17532301

ABSTRACT

The cholesterol 24-hydroxylase encoded by the gene CYP46 is expressed almost exclusively in central nervous system (CNS) neurons and catalyzes the formation of 24S-hydroxycholesterol (24S-OHC) from cholesterol. This conversion corresponds to a major pathway for excretion of excess cholesterol from the brain. There is a significant flux of another oxysterol, 27-hydroxycholesterol (27-OHC) from the circulation into the brain. Polymorphisms within the CYP46A1 gene have been associated with Alzheimer's disease (AD) incidence. In this study, we examined the effects of 24S-OHC and 27-OHC on the alpha- and beta-secretase activity in the human neuroblastoma cell line SH-SY5Y. Furthermore, we examined the effects of the two oxysterols on the levels of extra- and intracellular proteins of secreted APPalpha (sAPPalpha). Our findings suggest that 24S-OHC may exert a unique modulatory effect on APP processing and that this oxysterol increases the alpha-secretase activity as well as the alpha/beta-secretase activity ratio. The possibility is discussed that the ratio between 24S-OHC and 27-OHC is of importance for the generation of amyloid in the brain.


Subject(s)
Amyloid Precursor Protein Secretases/metabolism , Amyloid beta-Protein Precursor/metabolism , Gene Expression Regulation, Enzymologic/drug effects , Hydroxycholesterols/administration & dosage , Neuroblastoma/metabolism , Signal Transduction/drug effects , Cell Line, Tumor , Dose-Response Relationship, Drug , Humans
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