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1.
Biol. Res ; 52: 60, 2019. graf
Article in English | LILACS | ID: biblio-1100912

ABSTRACT

BACKGROUND: Recent studies have confirmed that RASAL1 has an antitumor effect in many cancers, but its functional role and the molecular mechanism underlying in colon cancer has not been investigated. RESULTS: We collected human colon cancer tissues and adjacent non-tumor tissues, human colon cancer cell lines LoVo, CaCo2, SW1116, SW480 and HCT-116, and normal colonic mucosa cell line NCM460. RT-qPCR was used to detect the RASAL1 level in the clinical tissues and cell lines. In LoVo and HCT-116, RASAL1 was artificially overexpressed. Cell viability and proliferation were measured using CCK-8 assays, and cell cycle was detected via PI staining and flow cytometry analysis. RASAL1 significantly inhibited the cell proliferation via inducing cell cycle arrest, suppressed cell cycle associated protein expression, and decreased the lipid content and inhibited the SCD1 expression. Moreover, SCD1 overexpression induced and downregulation repressed cell proliferation by causing cell cycle arrest. Additionally, luciferase reporter assays were performed to confirm the direct binding between SREBP1c, LXRα; and SCD1 promoter, we also demonstrated that RASAL1 inhibit SCD1 3'-UTR activity. RASAL1 inhibited tumor growth in xenograft nude mice models and shows inhibitory effect of SCD1 expression in vivo. CONCLUSION: Taken together, we concluded that RASAL1 inhibited colon cancer cell proliferation via modulating SCD1 activity through LXRα/SREBP1c pathway.


Subject(s)
Humans , Animals , Mice , Stearoyl-CoA Desaturase/metabolism , Colonic Neoplasms/pathology , GTPase-Activating Proteins/metabolism , Cell Proliferation/physiology , Sterol Regulatory Element Binding Protein 1/metabolism , Liver X Receptors/metabolism , Stearoyl-CoA Desaturase/genetics , Down-Regulation , GTPase-Activating Proteins/genetics , Cell Line, Tumor , Sterol Regulatory Element Binding Protein 1/genetics , Liver X Receptors/genetics
2.
Braz. J. Psychiatry (São Paulo, 1999, Impr.) ; 39(2): 95-103, Apr.-June 2017. tab
Article in English | LILACS | ID: biblio-844186

ABSTRACT

Objective: To study associations of cerebrovascular metabolism genotypes and haplotypes with age at Alzheimer’s disease dementia (AD) onset and with neuropsychiatric symptoms according to each dementia stage. Methods: Consecutive outpatients with late-onset AD were assessed for age at dementia onset and Neuropsychiatric Inventory scores according to Clinical Dementia Rating scores, apolipoprotein E gene (APOE) haplotypes, angiotensin-converting enzyme gene (ACE) variants rs1800764 and rs4291, low-density lipoprotein cholesterol receptor gene (LDLR) variants rs11669576 and rs5930, cholesteryl ester transfer protein gene (CETP) variants I422V and TaqIB, and liver X receptor beta gene (NR1H2) polymorphism rs2695121. Results: Considering 201 patients, only APOE-ɛ4 carriers had earlier dementia onset in multiple correlations, as well as less apathy, more delusions, and more aberrant motor behavior. Both ACE polymorphisms were associated with less intense frontally mediated behaviors. Regarding LDLR variants, carriers of the A allele of rs11669576 had less anxiety and more aberrant motor behavior, whereas carriers of the A allele of rs5930 had less delusions, less anxiety, more apathy, and more irritability. CETP variants that included G alleles of I422V and TaqIB were mostly associated with less intense frontally mediated behaviors, while severely impaired carriers of the T allele of rs2695121 had more anxiety and more aberrant motor behavior. Conclusion: Though only APOE haplotypes affected AD onset, cerebrovascular metabolism genotypes were associated with differences in several neuropsychiatric manifestations of AD.


Subject(s)
Humans , Male , Female , Middle Aged , Aged , Aged, 80 and over , Cerebrovascular Disorders/genetics , Cerebrovascular Disorders/metabolism , Alzheimer Disease/genetics , Alzheimer Disease/metabolism , Genotype , Apolipoproteins E/genetics , Linear Models , Cerebrovascular Disorders/physiopathology , Cross-Sectional Studies , Age of Onset , Gene Dosage , Alleles , Cholesterol Ester Transfer Proteins/genetics , Genetic Association Studies , Alzheimer Disease/physiopathology , Late Onset Disorders , Liver X Receptors/genetics , Lipoproteins, LDL/genetics , Neuropsychological Tests
3.
Cell Journal [Yakhteh]. 2017; 19 (1): 45-49
in English | IMEMR | ID: emr-185792

ABSTRACT

Objective: Liver X receptors [LXRs] are ligand-activated transcription factors of the nuclear hormonal receptor superfamily which modulate the expression of genes involved in cholesterol homeostasis. Hence, further unraveling of the molecular function of this gene may be helpful in preventing cardiovascular diseases


Materials and Methods: This experimental intervention study included twelve adult Wistar male rats [12-14 weeks old, 200-220 g] which were divided into the control [n=6] and training [n=6] groups. The training group received exercise on a motor-driven treadmill at 28 meters/minute [0% grade] for 60 minutes a day, 5 days a week for 4 weeks. Rats were sacrificed 24 hours after the last session of exercise. A portion of the liver was excised, immediately washed in ice-cold saline and frozen in liquid nitrogen for extraction of total RNA. Plasma was collected for high-density lipoprotein cholesterol [HDL-C], low-density lipoprotein cholesterol [LDL-C], total cholesterol [TC] and triglycerides [TG] measurements. All variables were compared by independent t test


Results: A significant increase in LXR alpha transcript level was observed in trained rats [P<0.01]. Plasma HDL-C concentration was also significantly higher [P<0.01] in trained rats. There was a significant decrease in the concentrations of LDL-C [P<0.01] and TC [P<0.02], and the ratios of TC/HDL-C [P<0.001] and LDL/HDL-C [P<0.002] in trained rats. However, the TG concentration was unchanged [P>0.05]


Conclusion: We found that endurance training induces significant elevation in LXR alpha gene expression and plasma HDL-C concentration resulting in depletion of the cellular cholesterol. Therefore, it seems that a contributor to the positive effects of exercise in cardiovascular disease prevention is through the expression of LXR alpha, which is a key step in reverse cholesterol transport


Subject(s)
Animals, Laboratory , Male , Liver X Receptors , Lipids , Cholesterol, HDL , Liver , Rats, Wistar
4.
Article in Chinese | WPRIM | ID: wpr-815063

ABSTRACT

OBJECTIVE@#To examine the expression of liver X receptor-β (LXR-β) in human gastric cancer tissue, and to explore the effect of GW3965, an agonist of LXRs, on proliferation of gastric cancer cell line SGC-7901.
@*METHODS@#The immunohistochemical assay was used to detect the expression of LXR-β, activating transcription factor 4 (ATF4) in gastric cancer tissues and the corresponding pericarcinoma tissues in 114 patients. Real-time quantitative PCR and Western blot were used to determine mRNA and protein levels of ATF4 and ATP-binding cassette 1 (ABCA1), one of the downstream target genes of LXRs, in SGC-7901 cells with or without GW3965 treatment. Cell counting kit-8 (CCK-8) assay was performed to detect cell proliferation. The expression of ATF4 was silenced by short hairpin RNA (shRNA).
@*RESULTS@#The expressions of LXR-β and ATF-4 were obviously down-regulated in the gastric cancer tissues than that in the corresponding pericarcinoma tissues (both P<0.05). Compared with the control cells, GW3965 treatment inhibited proliferation of SGC-7901 cells and up-regulated ATF4 and ABCA1 expressions (both P<0.05). Knockdown of ATF4 can reverse the antiproliferative effect of GW3965 on SGC-7901 cells.
@*CONCLUSION@#The expression of LXR-β is decreased in human gastric cancer tissues, and activation of LXRs by GW3965 could inhibit the proliferation of SGC-7901 cells via ATF4.


Subject(s)
Activating Transcription Factor 4 , Genetics , Metabolism , Benzoates , Pharmacology , Benzylamines , Pharmacology , Cell Line, Tumor , Cell Proliferation , Gene Expression Regulation, Neoplastic , Gene Silencing , Humans , Liver X Receptors , Orphan Nuclear Receptors , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , RNA, Small Interfering , Genetics , Stomach Neoplasms , Pathology , Up-Regulation
5.
Article in English | WPRIM | ID: wpr-308213

ABSTRACT

Nuclear receptor transcription factors are ligand-activated proteins that control various biological events from cell growth and development to lipid metabolism, and energy and glucose homeostasis. Nuclear receptors are important drug targets for metabolic diseases. Liver X receptors (LXRs) are nuclear receptor transcription factors that play essential roles in regulation of cholesterol, triglyceride, fatty acid, and glucose homeostasis. LXR-deficient mice have shown the association of LXR-signaling pathway dysfunction with several human pathologies including atherosclerosis, hyperlipidemia, Alzheimer's disease and cancer. Thus, LXRs are promising pharmacological targets for these diseases. Synthetic LXR agonists may lower cholesterol, but increase triglyceride and induce fatty liver. The naturally occurring LXR ligands, with moderate activity, may serve as nutraceuticals for prevention or treatment of the disorders, while minimizing potential side effects. In this review, recent advances in natural LXR modulators are summarized including agonist, antagonist and the modulator of LXR pathway.


Subject(s)
Animals , Biological Products , Pharmacology , Humans , Liver , Metabolism , Liver X Receptors , Orphan Nuclear Receptors , Physiology
6.
Article in Chinese | WPRIM | ID: wpr-312649

ABSTRACT

<p><b>OBJECTIVE</b>To investigate whether RNA interference (RNAi) of LXRα gene in donor rats with fatty liver improves liver graft function after transplantation.</p><p><b>METHODS</b>Fifty donor SD rats were fed a high-fat diet and 56% alcohol to induce macrovesicular steatosis exceeding 60% in the liver. The donor rats were injected via the portal veins with 7 × 10⁷ TU LXRα-RNAi-LV mixture (n=25) or negative control-LV (NC-LV) vector (n=25) 72 h before orthotopic liver transplantation. At 2, 24, and 72 h after the transplantation, the recipient rats were sacrificed to examine liver transaminases, liver graft histology, immunostaining (TUNEL), and protein and mRNA levels of LXRα.</p><p><b>RESULTS</b>Lentivirus-LXRα RNAi inhibited LXRα gene expression at both the mRNA and protein levels in the liver graft and reduced the expressions of SREBP-1c and CD36 as compared with the controls, resulting also in reduced fatty acid accumulation in the hepatocytes. The recipient rats receiving RNAi-treated grafts showed more obvious reduction in serum ALT, AST, IL-1β and TNF-α levels, and exhibited milder hepatic pathologies than the control rats after the transplantation. TUNEL assay demonstrated a significant reduction in cell apoptosis in LXRα-RNAi-LV-treated liver grafts, and the rats receiving treated liver grafts had a prolonged mean overall survival time.</p><p><b>CONCLUSION</b>LXRα-RNAi-LV treatment of the donor rats with fatty liver can significantly down-regulate LXRα gene expression in the liver graft and improve the graft function and recipient rat survival after liver transplantation.</p>


Subject(s)
Animals , Fatty Liver , Genetics , General Surgery , Gene Expression Regulation , Hepatocytes , Cell Biology , Lentivirus , Liver , Physiology , Liver Transplantation , Liver X Receptors , Orphan Nuclear Receptors , Genetics , RNA Interference , RNA, Messenger , Rats , Rats, Sprague-Dawley
7.
Chinese Medical Journal ; (24): 218-224, 2014.
Article in English | WPRIM | ID: wpr-318009

ABSTRACT

<p><b>BACKGROUND</b>Statin therapy has affected glucose homoeostasis of type 2 diabetes patients, which could be related with bile acids metabolism. Whether bile acid metabolism and the expression of farnesoid X receptor (FXR), liver X receptor-α (LXR-α) and sterol regulatory element-binding protein (Srebp)-1c is regulated by hyperglycemia, or whether simvastatin therapy led to higher glucose is related with down-regulated expression of FXR in diabetic rats remained unclear.</p><p><b>METHODS</b>Forty male Wistar rats were randomly divided into four groups: normal control rats, insulin resistance rats, diabetic model rats, and the late simvastatin induced diabetic rats. Normal control rats were fed with standard diet, others were fed with high-fat diet. Diabetic model rats were induced by a single intraperitoneal injection of streptozotocin (STZ). The late simvastatin induced diabetic rats started simvastatin administration after STZ induced diabetic model rats. Characteristics of fasting blood glucose (FPG), lipid files and total bile acids (TBAs) were measured and the oral glucose tolerance test (OGTT) was performed after overnight fasting at the eighth weekend. RNA and protein levels of FXR, LXR-α and Srebp-1c were tested by Western blotting and reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The insulin resistance rats showed higher glucose, lipid files and lower expression of FXR compared with normal control rats (P > 0.05). The diabetic model rats showed significantly higher glucose, lipid files, TBA and lower expression of FXR compared with insulin resistance rats (P < 0.05). The late simvastatin induced diabetic rats displayed higher glucose and TBA and lower expression of FXR compared with diabetic model rats (P < 0.05).</p><p><b>CONCLUSIONS</b>Changes in bile acid homeostasis, including the alterations of bile acid levels and bile acid receptors, are either a cause or a consequence of the metabolic disturbances observed during diabetic models. Statin therapy induced hyperglycemia may be related with FXR, SHP, LXR-α and Srebp-1 pathways.</p>


Subject(s)
Animals , Blood Glucose , Metabolism , Diabetes Mellitus, Experimental , Drug Therapy , Metabolism , Glucose Tolerance Test , Homeostasis , Insulin Resistance , Physiology , Liver X Receptors , Male , Orphan Nuclear Receptors , Metabolism , Rats , Rats, Wistar , Receptors, Cytoplasmic and Nuclear , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Simvastatin , Therapeutic Uses , Sterol Regulatory Element Binding Protein 1 , Metabolism
8.
São Paulo; s.n; s.n; 2013. 162 p. tab, graf, ilus.
Thesis in Portuguese | LILACS | ID: biblio-847171

ABSTRACT

Tiazolidinadionas (TZDs) são agentes sensibilizadores de insulina que agem por ligação ao receptor gama ativado por proliferador de peroxissomos (PPARγ). Elas têm apresentado efeitos cardioprotetores em humanos e propriedades anti-aterogênicas em modelos animais. Estudos in vitro têm sugerido que esses efeitos anti-aterogênicos da ativação de PPARγ ocorrem por inibição da expressão de genes pro-inflamatórios e por aumentar o efluxo de colesterol via ativação dos receptores LXR-ABCA1. Entretanto, vários efeitos colaterais são associados ao tratamento com as TZDs, tornando necessária a pesquisa por novos compostos desta classe. Neste estudo, 14 novas tiazolidina-2,4- dionas, que são TZDs modificadas por bioisosterismo, foram avaliadas quanto à expressão de fatores aterogênicos e inflamatórios em linhagens de macrófagos J774 e RAW 264.7 e em camundongos com deleção genética para o receptor de LDL (LDLr-/-). Após a avaliação da citotoxicidade em macrófagos, foram eleitas cinco TZDs, denominadas de GQ-11, GQ-97, GQ-177, GQ-145 e LYSO-7. Três destas TZDs (GQ- 145, GQ-177 e LYSO-7) aumentaram significativamente a expressão de RNAm dos fatores de transcrição PPARγ1, PPARγ2 e do receptor CD36, assim como também aumentaram a expressão gênica de ABCA1 em 2.9, 3.5 e 6.7 vezes, respectivamente. Em adição, estas TZDs diminuíram a expressão gênica de iNOS, COX2, VCAM e IL-6 associado a redução na produção de nitritos, mas apenas a LYSO-7 reduziu significativamente a expressão desses genes quando comparada à rosiglitazona (RSG), além de diminuir a expressão da proteína-1 quimiotática para monócitos (MCP-1). No estudo experimental, os camundongos LDLr-/- machos foram alimentados com dieta hipercolesterolêmica por 16 semanas e quatro semanas antes da eutanásia receberam os derivados tiazolidínicos (20 mg/kg/dia) por gavagem. GQ-177 inibiu a progressão da placa aterosclerótica associada à aumento nas concentrações plasmáticas de HDL-C, com elevação na expressão de ABCA1, e redução da via inflamatória CD40-CD40L. LYSO-7 também mostrou inibição da aterogênese associada à redução das concentrações plasmáticas de colesterol total e triacilgliceróis, com diminuição na interação entre CD40-CD40L e expressão de citocinas inflamatórias. A GQ-145 não alterou os níveis plasmáticos dos lipídeos, mas aumentou a expressão de todos os genes pró-aterogênicos e pró-inflamatórios. Adicionalmente, as vias de ativação destas novas TZDs também foram estudadas por ensaio de luciferase, como gene repórter. A GQ-177 induziu ativação de PPARγ e ligação ao seu domínio, enquanto a LYSO-7 estimulou ativação de PPARα e PPARδ. Portanto, conclui-se que as novas TZDs, especialmente a GQ-177 e a LYSO-7, podem apresentar propriedades ateroprotetoras associadas ao transporte reverso de colesterol e aos efeitos antiinflamatórios, e poderiam ser uma alternativa promissora para o tratamento da aterosclerose. Porém, estudos complementares são requeridos para caracterizar as vias de sinalização intracelular, visto que as duas demonstraram ativar diferentes isotipos do fator de transcrição PPAR


Thiazolidinediones (TZDs) are insulin-sensitizing agents that act by binding to peroxisome proliferator-activated receptor-γ (PPARγ). They have been demonstrated to possess cardioprotective effects in humans and antiatherogenic properties in animal models. In vitro studies have also suggested that these antiatherogenic effects of PPARγ activation occur by inhibiting the inflammatory gene expression and by increasing cholesterol efflux via LXR-ABCA1 activation. However, several side effects are associated with TZDs treatment making necessary the search for new compounds. In this study, 14 new thiazolidine-2,4-diones, modified TZDs by bioisosterism, were tested for aterogenic and inflammtary factors in RAW 264.7 macrophages and in low-density lipoprotein receptor-deficient mice. After the citotoxicity evaluation in RAW 264.7 macrophages the TZDs named GQ-11, GQ-97, GQ-177, GQ-145 e LYSO-7 were selected for this study. Three of these TZDs (GQ-177, GQ-145 and LYSO-7) significantly increased the expression of PPARγ1, PPARγ2 and CD36 mRNA, and enhanced the expression of ABCA1 mRNA in 2.9, 3.5 and 6.7 fold, respectively. Moreover, they also significantly decreased the expression of iNOS, COX2, VCAM and IL-6 mRNA in relation to control, and these results are associated to reduction on nitrits concentration. In addition, LYSO-7 significantly reduced the expression of these genes when compared to rosiglitazone, and decreased expression of MCP1 mRNA. In the experimental study, male LDLr-/- mice were fed an atherogenic diet containing 0.5% cholesterol for 16 weeks, and 4 weeks before euthanasia they received TZDs (20mg/kg/ per day) by gavage. GQ-177 treatment inhibited progression of atherosclerotic plaque associated to increased plasma concentrations of HDL-C, with enhance of ABCA1 expression and reduction on CD40-CD40L interaction. LYSO-7 treatment also showed inhibition of the atherogenesis associated to decreased plasma concentrations of total cholesterol and TAG, with reduction on CD40-CD40L pathway and inflammatory cytokines expression.GQ-145 did not alter the lipid plasma levels and increased the expression of all pro-atherogenic and pro-inflammatory genes. Furthermore, the activation of PPARs has also been studied, by luciferase assay as reporter gene. GQ-177 induced activation of PPARγ, whereas LYSO-7 stimulated activation of PPARα and PPARß/δ. Altogether, our data suggest that the new TZDs derivatives, specially GQ- 177 and LYSO-7, may have atheroprotective properties associated with the reverse cholesterol transport and anti-inflammatory effects, and could be a promising alternative for the treatment of atherosclerosis. However, further studies are warranted in order to characterize the pathways of intracellular signaling since both have demonstrated to activate different isotypes of PPAR


Subject(s)
Animals , Male , Mice , Atherosclerosis/pathology , Luciferases/pharmacology , Cell Death , Cell Survival , Liver X Receptors/analysis , Peroxisome Proliferator-Activated Receptors , PPAR gamma
9.
Chinese Medical Journal ; (24): 306-310, 2013.
Article in English | WPRIM | ID: wpr-331275

ABSTRACT

<p><b>BACKGROUND</b>ABCA7 is a member of the ABCA subfamily that shows a high degree of homology to ABCA1 and, like ABCA1, mediates cellular cholesterol and phospholipid release by apolipoproteins when transfected in vitro. However, expression of ABCA7 has been shown to be downregulated by increased cellular cholesterol while ABCA1 was upregulated.</p><p><b>METHODS</b>The underlying mechanism for this effect was examined in ABCA1 or ABCA7-transfected HEC293. Lipid content in the medium and cells was determined by enzymatic assays. Gene expression was quantitated by real time PCR, and protein content was determined by Western blotting.</p><p><b>RESULTS</b>While ABCA7 mRNA was decreased by 25-hydroxycholesterol treatment, ABCA1 was apparently increased. Treatment with the synthetic LXR agonist T0901317 (T09) upregulated ABCA1 expression and apoAI-mediated cellular lipid release in ABCA1-transfected HEC293 cells, but ABCA7 expression and cellular lipid release in ABCA7-transfected HEC293 cells showed no obvious changes.</p><p><b>CONCLUSION</b>The ABCA7 gene is regulated by sterol in a direction opposite to that of ABCA1.</p>


Subject(s)
ATP Binding Cassette Transporter 1 , Genetics , Physiology , ATP-Binding Cassette Transporters , Genetics , Physiology , Amino Acid Sequence , Apolipoprotein A-I , Physiology , Gene Expression Regulation , HEK293 Cells , Humans , Hydrocarbons, Fluorinated , Pharmacology , Hydroxycholesterols , Pharmacology , Lipid Metabolism , Liver X Receptors , Molecular Sequence Data , Orphan Nuclear Receptors , Sulfonamides , Pharmacology
10.
Chinese Medical Journal ; (24): 3539-3542, 2012.
Article in English | WPRIM | ID: wpr-256699

ABSTRACT

<p><b>BACKGROUND</b>The gradually increasing changes in a human hyperlipidemic diet along with chronic stress might play an important role in the increased numbers of fatty liver. This study investigated the effects of Ilex asprella root decoction on related genes of lipid metabolism in chronic stress in hyperlipidemic fatty liver in rats.</p><p><b>METHODS</b>Forty-eight male Wistar rats were randomly divided into four groups: normal control group, model control group, simvastatin group, and Ilex asprella root group. To establish chronic stress and hyperlipidemic fatty liver models in rats, the levels of serum lipids, glucose, liver index, insulin (INS), insulin resistant (IR) index, adiponectin, superoxide dismutase (SOD), glutathione peroxidase (GSH-pX), glutathione (GSH), liver X receptor (LXR), and sterol responsive element binding protein (SREBP)-1c in rats were measured.</p><p><b>RESULTS</b>When compared to the normal control group, the levels of serum lipids, glucose, liver index, INS, IR index, and GSH in the model control group significantly increased (P < 0.01). The protein levels of LXRα and SREBP-1c increased (P < 0.05), and the serum adiponectin and the SOD and GSH-pX decreased significantly (P < 0.01). When compared to the model control group, the levels of serum lipids, glucose, liver index, INS, IR index, SOD, and GSH-pX in the simvastatin group and Ilex asprella root group increased in varying degrees (P < 0.01 or 0.05); the serum adiponectin and GSH decreased (P < 0.05), while the protein levels of LXRα and SREBP-1c decreased in varying degrees (P < 0.01 or 0.05). When compared to the simvastatin group, the IR index and protein levels of LXRα in the Ilex asprella root group decreased (P < 0.05), and the serum adiponectin and SOD increased (P < 0.05).</p><p><b>CONCLUSION</b>The Ilex asprella root decoction has some protective effects on regulating the related genes of lipid metabolism caused by chronic stress and hyperlipidemic fatty liver in rats.</p>


Subject(s)
Animals , Fatty Liver , Drug Therapy , Metabolism , Hyperlipidemias , Drug Therapy , Metabolism , Ilex , Chemistry , Lipid Metabolism , Lipid Peroxidation , Liver X Receptors , Male , Orphan Nuclear Receptors , Genetics , Plant Extracts , Chemistry , Plant Roots , Chemistry , Rats , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Sterol Regulatory Element Binding Protein 1 , Genetics
11.
Chinese Journal of Cardiology ; (12): 161-165, 2012.
Article in Chinese | WPRIM | ID: wpr-275083

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of liver X receptors (LXRs) on endothelin-1 (ET-1) induced murine HL-1 cardiomyocytes hypertrophy.</p><p><b>METHODS</b>Cultured murine HL-1 cardiomyocytes were divided into four experiment groups: (1) CONTROL GROUP:treated with DMSO; (2) T0901317 group:treated with LXRs agonist T0901317 (1 µmol/L); (3) ET-1 group:treated with ET-1 (1 nmol/L); (4) T0901317 + ET-1 group:treated with T0901317 (1 µmol/L) for 8 hours, then treated with ET-1 (1 nmol/L). Twenty-four hours later, immunofluorescent staining was performed on HL-1 cells, the surface area of HL-1 cells was analyzed with NIH Image J software, and the synthetic rate of protein in HL-1 cells was detected by (3)H-leucine incorporation. The mRNA level of atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MyHC) was measured by quantitative realtime PCR. The effect of T0901317 on mRNA expression of ANP was also detected after LXRs gene silencing.</p><p><b>RESULTS</b>The surface area of HL-1 cells, mRNA expression of ANP and β-MyHC, and (3)H-leucine incorporation in ET-1 group were 2.00 ± 0.29, 1.98 ± 0.47, 2.13 ± 0.39 and 1.79 ± 0.17, respectively, which were significantly higher than those of control group (1.00 ± 0.26, 1.00 ± 0.21, 1.00 ± 0.31 and 1.00 ± 0.03, respectively, all P < 0.05). Compared with ET-1 group, the surface area of HL-1 cells, mRNA expression of ANP and β-MyHC, and (3)H-leucine incorporation were significantly decreased in T0901317 + ET-1 group (1.24 ± 0.25, 1.19 ± 0.21, 1.48 ± 0.27 and 1.15 ± 0.11, respectively, all P < 0.05). After inhibition of LXRα/β expression in HL-1 cardiomyocytes using the specific siRNAs, the mRNA expression of ANP in T0901317 + ET-1 group was 1.78 ± 0.05, which was similar as that in ET-1 group (1.94 ± 0.17, P > 0.05).</p><p><b>CONCLUSION</b>T0901317, an agonist of LXRs, could inhibit ET-1 induced cardiac hypertrophy in vitro, and LXR ligand-mediated inhibition on ANP mRNA expression by T0901317 is receptor dependent.</p>


Subject(s)
Animals , Cardiomegaly , Metabolism , Cell Line , Endothelin-1 , Metabolism , Hydrocarbons, Fluorinated , Pharmacology , Liver X Receptors , Mice , Myocytes, Cardiac , Metabolism , Orphan Nuclear Receptors , Metabolism , Signal Transduction , Sulfonamides , Pharmacology
12.
Acta Pharmaceutica Sinica ; (12): 427-433, 2012.
Article in Chinese | WPRIM | ID: wpr-323024

ABSTRACT

Liver X receptor (LXR), a member of the superfamily of nuclear receptors, plays an important role in the activation of transcription factors involved in cholesterol metabolism, glucose homeostasis inflammation and lipogenesis. It is shown that LXR agnoists have the potentiality to be used as drugs for the prevention and treatment of atherosclerosis, which is its best investigated therapeutic indication. There are many compounds being studied in preclinical evaluation and biological assay. This paper will review briefly the LXR agonists in recent years.


Subject(s)
ATP-Binding Cassette Transporters , Metabolism , Amines , Chemistry , Pharmacology , Animals , Atherosclerosis , Drug Therapy , Metabolism , Benzimidazoles , Chemistry , Pharmacology , Cholesterol , Pharmacology , Glucose , Pharmacology , Humans , Lipid Metabolism , Lipogenesis , Liver X Receptors , Orphan Nuclear Receptors , Physiology , Quinolines , Chemistry , Pharmacology , Sterol Regulatory Element Binding Protein 1 , Metabolism
13.
Chinese Journal of Cardiology ; (12): 723-728, 2012.
Article in Chinese | WPRIM | ID: wpr-326432

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of liver X receptor (LXR) agonist on adipose-derived mesenchymal stem cells (AD-MSCs) implantation into infarcted hearts of mice.</p><p><b>METHOD</b>AD-MSC(Fluc+) which stably expressed firefly luciferase (Fluc) were isolated from β-actin-Fluc transgenic mice and characterized by flow cytometry. Male FVB mice were randomly allocated into the following four groups (n = 10 each): (1) sham group; (2) MI + PBS group; (3) MI + AD-MSC(Fluc+) group; (4) MI + AD-MSC(Fluc+) + LXR agonist (T0901317) group. AD-MSC(Fluc+) or PBS were injected intramyocardial into peri-infarcted region of mice heart after permanent left anterior descending (LAD) artery ligation. Bioluminescence imaging (BLI) was performed for quantification of injected cells retention and survival. Cardiac function was evaluated by echocardiography.</p><p><b>RESULTS</b>The AD-MSC(Fluc+) were positive for CD44 and CD90 by flow cytometry. BLI evidenced the firefly luciferase expression of AD-MSC(Fluc+) which was positively correlated with cell numbers (r(2) = 0.98). The results of BLI in vivo revealed that LXR agonist could improve the survival of AD-MSC(Fluc+) at day 7, 14 and 21 after transplantation compared with AD-MSC(Fluc+) alone group. Cardiac function was further improved in combination therapy group compared with AD-MSC(Fluc+) alone group (P < 0.05).</p><p><b>CONCLUSIONS</b>LXR agonist T0901317 can improve the retention and survival of intramyocardial injected AD-MSC(Fluc+) post-MI, and the combination therapy of T0901317 and AD-MSC(Fluc+) has a synergetic effect on improving cardiac function in this model.</p>


Subject(s)
Animals , Hydrocarbons, Fluorinated , Therapeutic Uses , Liver X Receptors , Male , Mesenchymal Stem Cell Transplantation , Methods , Mice , Mice, Transgenic , Myocardial Infarction , Mortality , General Surgery , Orphan Nuclear Receptors , Sulfonamides , Therapeutic Uses , Treatment Outcome
14.
Chinese Journal of Hepatology ; (12): 768-773, 2011.
Article in Chinese | WPRIM | ID: wpr-239330

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of Hepatitis B Virus X Protein (HBx) on the expression of lipid metabolism-related genes and its role in pathogenesis of hepatocyte fatty degeneration.</p><p><b>METHODS</b>Hepatitis B Virus X gene eukaryon expression vector pIRES2-eGFP-HBx was transfected into HepG2 cells to establish HepG2/HBx cell model for HBx expression. HepG2 cells transfected with pIRES2-eGFP (HepG2/pIRES2 cell) and non-transfected were used as controls. At 24, 48 and 72 hours after transfection, the expression of green fluorescent protein (GFP) was observed by fluorescence microscope and the triglyceride(TG) content was detected. RT-PCR and Western blot were applied to detect the levels of sterol regulatory element binding protein-1 (SREBP-1), liver x receptor alpha (LXRalpha) mRNA and the levels of HBx, LXRalpha and fatty acid synthase (FAS) protein. At 24, 48 and 72 hours after transfection, the expression of GFP was found in HepG2/HBx and HepG2/pIRES2 cells, and increased gradually. The expression of HBx was detected only in HepG2/HBx cells, and was increased with time after transfection (F = 32.21, P less than 0.01). These suggested successful obtaining of HepG2-HBx cell model for HBx expression.</p><p><b>RESULTS</b>At 24h, 48h and 72h after transfection, the expression levels of LXRalpha mRNA (0.386+/-0.055, 0.505+/-0.071, 0.649+/-0.058 ) and SREBP-1 mRNA (0.395+/-0.055, 0.548+/-0.047, 0.795+/-0.058), as well as the levels of LXRalpha protein(0.178+/-0.036, 0.263+/-0.047, 0.347+/-0.058) and FAS protein(0.436+/-0.055, 0.608+/-0.053, 0.827+/-0.046) in HepG2-HBx group were dramatically higher than those in the controls at the same time points (all P less than 0.05/0.01), and were gradually increased with time (all P less than 0.05/0.01). A positive correlationship was observed between HBX protein level and the LXRalpha, SREbP-1 mRNA and LXRalpha, FAS protein levels. The difference of TG content between HepG2/HBx group and control groups was not statistically significant (P more than 0.05).</p><p><b>CONCLUSIONS</b>HBx-LXRalpha-SREBP-1/FAS pathway suggested regulating transcription and expression of lipid metabolism-related genes, which might be one of the important molecular mechanism causing hepatocyte fatty degeneration.</p>


Subject(s)
Carcinoma, Hepatocellular , Metabolism , Fatty Acid Synthase, Type I , Metabolism , Hep G2 Cells , Humans , Lipid Metabolism , Genetics , Liver Neoplasms , Metabolism , Liver X Receptors , Orphan Nuclear Receptors , Metabolism , Sterol Regulatory Element Binding Protein 1 , Metabolism , Trans-Activators , Genetics , Transfection
15.
Article in Chinese | WPRIM | ID: wpr-332558

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of liver X receptor agonist T0901317 on transforming growth factor-β1 (TGF-β1)-induced expression of α-smooth muscle actin (α-SMA) in normal human lung fibroblasts.</p><p><b>METHODS</b>Primary normal human lung fibroblast isolated from the lung specimens of lung cancer patients by explant culture technique were identified with immunostaining for vimentin and keratin. The cells in passages 4 to 10 were treated with T0901317 and/or TGF-β1, and RT-PCR, Western blotting and immunofluorescence assay were used to detect α-SMA expression in the fibroblasts.</p><p><b>RESULTS</b>Lung fibroblast expressed vimentin but not keratin. The results of RT-PCR, Western blotting and immunofluorescence assay all showed that normal human lung fibroblasts constitutively expressed α-SMA under baseline condition, and TGF-β1 at 5 ng/ml induced a significant upregulation of α-SMA both at the mRNA and protein levels. Liver X receptor agonist T0901317 (5 µg/ml) significantly inhibited TGF-β1-induced upregulation of α-SMA expression.</p><p><b>CONCLUSION</b>Liver X receptor agonist T0901317 can inhibit the upregulation of α-SMA in normal human lung fibroblasts induced by TGF-β1, suggesting the potential value of liver X receptor agonist in the treatment of lung fibrosis.</p>


Subject(s)
Actins , Metabolism , Cells, Cultured , Female , Fibroblasts , Metabolism , Humans , Hydrocarbons, Fluorinated , Pharmacology , Liver X Receptors , Lung , Cell Biology , Middle Aged , Orphan Nuclear Receptors , RNA, Messenger , Genetics , Sulfonamides , Pharmacology , Transforming Growth Factor beta1 , Pharmacology
16.
Chinese Journal of Hepatology ; (12): 542-546, 2011.
Article in Chinese | WPRIM | ID: wpr-330702

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the relationship of NOR-1 with the inhibition of inflammatory reaction in mice Kupffer cells (KCs) induced by lipopolysaccharide (LPS) via liver X receptor alpha (LXR alpha).</p><p><b>METHODS</b>KCs from male KM mice were isolated by density gradient centrifugation, incubated and then randomly assigned to three groups: control group, LPS treated group and LPS+T0901317 treated group.</p><p><b>RESULTS</b>The mRNA and protein expressions of LXR alpha and NOR-1 in each group were determined by RT-PCR, immunofluorescent assay and western blot, respectively. The densities of TNF alpha and IL-10 in supernatants were evaluated by enzyme linked immunosorbent assay (ELISA). The mRNA and protein expression levels of LXR alpha in LPS + T0901317 group were the highest as compared to the other two groups (0.748+/-0.072 and 1.217+/-0.133 respectively), The mRNA and protein expression levels of NOR-1 in LPS+ T0901317 group were the highest as compared to the other two groups (2.726+/-0.065 and 0.842+/-0.058 respectively). The densities of supernatant TNF alpha in LPS group and IL-10 in LPS+T0901317 group were the highest [(450.89+/-78.52) ng/L and (537.41+/-36.41) ng/L respectively].</p><p><b>CONCLUSIONS</b>Promoting the expression of LXR alpha in KCs can elevate the NOR-1 expression and then inhibit inflammatory reaction.</p>


Subject(s)
Animals , Cells, Cultured , DNA-Binding Proteins , Metabolism , Inflammation , Metabolism , Interleukin-10 , Metabolism , Kupffer Cells , Metabolism , Liver X Receptors , Male , Mice , Mice, Inbred Strains , Nerve Tissue Proteins , Metabolism , Orphan Nuclear Receptors , Metabolism , Receptors, Steroid , Metabolism , Receptors, Thyroid Hormone , Metabolism , Tumor Necrosis Factor-alpha , Metabolism
17.
Article in Chinese | WPRIM | ID: wpr-268829

ABSTRACT

<p><b>OBJECTIVE</b>To explore the possible mechanism of the inhibitory effect of liver X receptor alpha (LXRalpha) on lipopolysaccharide (LPS)-induced inflammation in mouse Kupffer cells (KCs).</p><p><b>METHODS</b>The KCs isolated from the liver of male KM mice and cultured in RPMI 1640 containing 20% FBS for 24 h were divided into control, LPS, T0901317, and LPS+T0901317 groups with corresponding treatments. The expressions of LXRalpha, interferon regulatory factor 3 (IRF3) and glucocorticoid receptor interacting protein 1 (GRIP1) in the KCs were detected by Western blotting. The levels of interferon beta (IFNbeta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in the supernatant were detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The level of LXRalpha protein was highest in T0901317 group and lowest in LPS group, and was significantly higher in LPS+T0901317 group than in LPS group but lower than in T0901317 group (P<0.05). The levels of IRF3 and GRIP1 protein were the highest in LPS group, and significantly lowered by T0901317 treatment (P<0.05). The expression of IRF3 and GRIP1 proteins in LPS group and LPS+ T0901317 group were significantly higher than those in the control and T0901317 groups (P<0.05). The concentration of IFN-beta was significantly higher in LPS group than in the control and T0901317 group (P<0.05), and decreased in LPS+T0901317 group in comparison with that in LPS group (P<0.05). IFN-beta was the lowest in T0901317 group. The levels of TNF-alpha and IL-1beta were the highest in LPS group (P<0.05), and comparable between the other 3 groups (P>0.05).</p><p><b>CONCLUSION</b>Pre-treatment with T0901317 before LPS stimulation can suppress the expressions of IRF3 and GRIP1 to inhibit the inflammation and hence Kupffer cell activation.</p>


Subject(s)
Animals , Cells, Cultured , Hydrocarbons, Fluorinated , Pharmacology , Inflammation , Interferon Regulatory Factor-3 , Metabolism , Kupffer Cells , Cell Biology , Metabolism , Lipopolysaccharides , Pharmacology , Liver X Receptors , Male , Mice , Nuclear Receptor Coactivator 2 , Metabolism , Orphan Nuclear Receptors , Physiology , Sulfonamides , Pharmacology
18.
Chinese Journal of Biotechnology ; (12): 354-359, 2009.
Article in Chinese | WPRIM | ID: wpr-286705

ABSTRACT

In order to examine the role of astacene on mice body development and the expression of energy metabolism related genes in mice, we treated mice (Kunming white) and primary culture of mouse muscle cells with astacene of higher and lower concentration. Then the total mRNA was extracted from the muscle tissue and cells respectively, and the mRNA levels of UCP3 and LXRalpha were detected by RT-PCR in all the samples. Compared with the control group, the body weight of mice in high concentrations of astacene group grown slowly, and the expressions of UCP3 genes decreased significantly in muscle tissue of the 10th day and the 30th day as well as the cells of treated for 24 h (P<0.05). The expression of LXRalpha gene increased significantly in all samples (P<0.05) and reached its peak at 72 h (P<0.01). With the treatment of lower concentration of astacene, the expressions of UCP3 and LXRalpha gene mRNA in muscle tissue did not alter much, but in muscle cells treated for 24 h, the mRNA level of UCP3 gene decreased significantly (P<0.05), and LXRalpha gene increased significantly (P<0.05). The results suggest that astacene has a role in regulating the energy use in mice muscle.


Subject(s)
Animals , Carotenoids , Pharmacology , Cells, Cultured , Dose-Response Relationship, Drug , Energy Metabolism , Genetics , Ion Channels , Genetics , Metabolism , Liver X Receptors , Male , Mice , Mitochondrial Proteins , Genetics , Metabolism , Muscle, Skeletal , Cell Biology , Metabolism , Orphan Nuclear Receptors , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Random Allocation , Uncoupling Protein 3
19.
Chinese Journal of Cardiology ; (12): 1119-1123, 2009.
Article in Chinese | WPRIM | ID: wpr-323898

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of liver X receptors (LXR) in hypertrophic myocardium and the effect of LXR agonist T0901317 on angiotensin II (AngII) induced cardiomyocyte hypertrophy.</p><p><b>METHODS</b>Transverse aortic coarctation (TAC) or sham operation were performed in 2-month-old wide type mice (C57/B6). Two weeks later, the expression of LXR in myocardium was detected by quantitative real-time PCR analysis and Western blot analysis. The effect of LXR agonist T0901317 on AngII-induced hypertrophy in cultured neonatal rat cardiomyocytes was also assessed.</p><p><b>RESULTS</b>Quantitative real-time PCR analysis and Western blot analysis showed that LXRalpha but not LXRbeta expression was upregulated post TAC both at mRNA and protein levels (All P < 0.05). AngII induced increased [(3)H] leucine incorporation and cardiomyocyte hypertrophy were significantly reduced by T0901317 in a dose-dependent manner (P < 0.05). T0901317 also dose-dependently inhibited atrial natriuretic peptide (ANP) gene expression in cardiomyocytes (P < 0.05).</p><p><b>CONCLUSION</b>Our findings strongly suggest that LXR is a potent mediator of cardiomyocyte hypertrophy and LXR activation could attenuate AngII induced cardiomyocyte hypertrophy in vitro.</p>


Subject(s)
Angiotensin II , Pharmacology , Animals , Animals, Wild , Cells, Cultured , Hydrocarbons, Fluorinated , Pharmacology , Liver X Receptors , Male , Mice , Mice, Inbred C57BL , Myocytes, Cardiac , Pathology , Orphan Nuclear Receptors , Metabolism , Sulfonamides , Pharmacology
20.
Article in Chinese | WPRIM | ID: wpr-325571

ABSTRACT

<p><b>OBJECTIVE</b>Preparing rat model of non-alcoholic fatty liver disease by fat-rich diet to observe the expression and the role of LXR-alpha in rat nonalcoholic fatty liver disease.</p><p><b>METHODS</b>Thirty-six SD rats were randonmized into basic diet-control group and high-fat diet group. Each of the two groups was subdivided into 3 subgroups (4, 8, 12 weeks). Changes in animal weight, liver exponent and the level of TG and TC in serum and liver were observed dynamically. Meanwhile,the expression of hepatocyte LXR-alpha and SREBP-1c were assayed by Reverse transcript-polymerase chain reaction at 4, 8, 12 weeks. The level of steatosis was observed under light microscope after haematoxylon-eosin (HE) staening.</p><p><b>RESULTS</b>Compared with control group, body weight, liver exponent, TG and Tc in serum and liver were increased dynamically in model groups. Compared with control group, the mRNA of LXR-a and SREBP-1c were obviously increased dynamically in model groups (P < 0.05) .</p><p><b>CONCLUSION</b>The increase of LXR-alpha and SREBP-1c in liver may be concered with energy disorder and closely associated with the activity of inflammation and the severity of the liver damage in NAFLD rats.</p>


Subject(s)
Animals , Disease Models, Animal , Fatty Liver , Genetics , Metabolism , Humans , Liver X Receptors , Male , Orphan Nuclear Receptors , Genetics , Metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , Sterol Regulatory Element Binding Protein 1 , Genetics , Metabolism
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