RESUMEN
In the present study the ability of supercritical carbon dioxide (SCO2) extracts of M. longifolia L. leaves to modulate low-density lipoprotein receptor (LDLR) and proprotein convertase subtilisin/kexin type 9 (PCSK9) expression was evaluated in cultured human hepatoma cell lines Huh7 and HepG2. Two SCO2 extracts, one oil (ML-SCO2) and a semisolid (MW-SCO2), were subjected to detailed chemical characterization by mono- and bidimensional nuclear magnetic resonance (1D, 2D-NMR), gas chromatography coupled with mass spectrometry (GC-MS) and liquid chromatography coupled with mass spectrometry (LC-MS). Chemical analysis revealed significant amounts of fatty acids, phytosterols and terpenoids. ML-SCO2 was able to induce LDLR expression at a dose of 60 µg/mL in HuH7 and HepG2 cell lines. Furthermore, ML-SCO2 reduced PCSK9 secretion in a concentration-dependent manner in both cell lines. Piperitone oxide, the most abundant compound of the volatile constituent of ML-SCO2 (27% w/w), was isolated and tested for the same targets, showing a very effective reduction of PCSK9 expression. The overall results revealed the opportunity to obtain a new nutraceutical ingredient with a high amount of phytosterols and terpenoids using the SCO2 extraction of M. longifolia L., a very well-known botanical species used as food. Furthermore, for the first time we report the high activity of piperitone oxide in the reduction of PCSK9 expression.
Asunto(s)
Dióxido de Carbono/química , Monoterpenos Ciclohexánicos , Mentha/química , Extractos Vegetales/química , Proproteína Convertasa 9/biosíntesis , Receptores de LDL/biosíntesis , Monoterpenos Ciclohexánicos/química , Monoterpenos Ciclohexánicos/aislamiento & purificación , Monoterpenos Ciclohexánicos/farmacología , Células Hep G2 , HumanosRESUMEN
Ascorbic acid, a water-soluble antioxidant, regulates various biological processes and is thought to influence cholesterol. However, little is known about the mechanisms underpinning ascorbic acid-mediated cholesterol metabolism. Here, we determined if ascorbic acid can regulate expression of proprotein convertase subtilisin/kexin 9 (PCSK9), which binds low-density lipoprotein receptor (LDLR) leading to its intracellular degradation, to influence low-density lipoprotein (LDL) metabolism. At cellular levels, ascorbic acid inhibited PCSK9 expression in HepG2 and Huh7 cell lines. Consequently, LDLR expression and cellular LDL uptake were enhanced. Similar effects of ascorbic acid on PCSK9 and LDLR expression were observed in mouse primary hepatocytes. Mechanistically, ascorbic acid suppressed PCSK9 expression in a forkhead box O3-dependent manner. In addition, ascorbic acid increased LDLR transcription by regulating sterol regulatory element-binding protein 2. In vivo, administration of ascorbic acid reduced serum PCSK9 levels and enhanced liver LDLR expression in C57BL/6J mice. Reciprocally, lack of ascorbic acid supplementation in L-gulono-γ-lactone oxidase deficient (Gulo-/-) mice increased circulating PCSK9 and LDL levels, and decreased liver LDLR expression, whereas ascorbic acid supplementation decreased PCSK9 and increased LDLR expression, ameliorating LDL levels in Gulo-/- mice fed a high fat diet. Moreover, ascorbic acid levels were negatively correlated to PCSK9, total and LDL levels in human serum samples. Taken together, these findings suggest that ascorbic acid reduces PCSK9 expression, leading to increased LDLR expression and cellular LDL uptake. Thus, supplementation of ascorbic acid may ameliorate lipid profiles in ascorbic acid-deficient species.
Asunto(s)
Ácido Ascórbico/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Proproteína Convertasa 9/biosíntesis , Receptores de LDL/biosíntesis , Animales , Células Hep G2 , Humanos , L-Gulonolactona Oxidasa/genética , L-Gulonolactona Oxidasa/metabolismo , Lipoproteínas LDL/genética , Lipoproteínas LDL/metabolismo , Ratones , Ratones Noqueados , Proproteína Convertasa 9/genética , Receptores de LDL/genéticaRESUMEN
Statins are the first-line treatment for hypercholesterolemic patients. Herein, the effects of three statins on complex formation between proprotein convertase subtilisin-kexin 9 (PCSK9) and the low density lipoprotein receptor (LDLR), a critical step for the PCSK9-dependent degradation of LDLR in the lysosome, were examined. Human hepatocyte-like C3A cells grown in control (containing 10% fetal bovine serum) or MITO+ (supplemented with BD™ MITO + serum extender) medium were also treated with atorvastatin (Atorv), lovastatin (Lov), or pravastatin (Prav) for 24 h. RNA and protein expression studies and determinations of PCSK9/LDLR complex formation were performed. As expected, the statins increased the expression of PCSK9 and LDLR independently of the medium employed. Interestingly, Atov and Lov caused increases in PCSK9/LDLR complex formation, whereas Prav decreased complex formation when compared to cells treated without drugs. These results may explain why Prav works better for statin intolerant patients than other statins such as Atorv and Lov.
Asunto(s)
Atorvastatina/farmacología , Lovastatina/farmacología , Inhibidores de PCSK9 , Pravastatina/farmacología , Proproteína Convertasa 9/biosíntesis , Receptores de LDL/antagonistas & inhibidores , Receptores de LDL/biosíntesis , Células Cultivadas , Humanos , Relación Estructura-ActividadRESUMEN
OBJECTIVE: We have previously shown that CLA induces regression of pre-established atherosclerotic lesions in apoE(-/-) mice. CLA is a known ligand of peroxisome proliferator activated receptors (PPARs) and it is postulated that CLA mediates its atheroprotective effects through activation of PPARs. Earlier work in our group identified the monocyte/macrophage cell as the primary cellular target of CLA. In this study we identified novel genes regulated by CLA during the regression of atherosclerosis and characterised a role for one of these, SorLA. SorLA is a member of the vacuolar protein sorting 10 protein (Vps10p) domain receptor family, which has structural homology with the LDLR family. METHODS AND RESULTS: Expression of SorLA was identified with its Vps10p family member Sort1 by transcriptomic analysis of murine aorta following CLA-induced regression of atherosclerosis. Decreased expression of both receptors was confirmed by real-time PCR in the aorta of CLA-supplemented mice. SorLA protein expression was predominantly localised to monocyte/macrophage cells in the vasculature by immunohistochemistry. CLA and the PPAR-γ agonists, troglitazone, and 15-deoxy-prostaglandin (PG) J(2), decreased protein and RNA expression of SorLA in THP-1 monocytes; while pre-treatment with a PPAR-γ antagonist established a PPAR-γ dependent role for CLA regulation of SorLA. CLA inhibits monocyte migration. Consistent with a role for SorLA in mediating this response, overexpression of SorLA increased migration of THP-1 monocytes to monocyte chemoattractant protein-1 with a coincident increase in UPAR expression. CONCLUSION: CLA may mediate its atheroprotective effects in part through reduced expression of SorLA and a resulting inhibition of monocyte migration in vitro.
Asunto(s)
Ácidos Linoleicos Conjugados/farmacología , Proteínas de Transporte de Membrana/fisiología , Monocitos/fisiología , Receptores de LDL/fisiología , Animales , Aterosclerosis/prevención & control , Movimiento Celular/efectos de los fármacos , Quimiocina CCL2/farmacología , Cromanos/farmacología , Humanos , Proteínas de Transporte de Membrana/biosíntesis , Ratones , Monocitos/efectos de los fármacos , PPAR gamma/antagonistas & inhibidores , PPAR gamma/metabolismo , Receptores de LDL/biosíntesis , Receptores del Activador de Plasminógeno Tipo Uroquinasa/metabolismo , Tiazolidinedionas/farmacología , TroglitazonaRESUMEN
Zingiber officinale has been used to control lipid disorders and reported to possess remarkable cholesterol-lowering activity in experimental hyperlipidaemia. In the present study, the effect of a characterized and standardized extract of Zingiber officinale on the hepatic lipid levels as well as on the hepatic mRNA and protein expression of low-density lipoprotein (LDL) receptor and 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase was investigated in a high-fat diet-fed rat model. Rats were treated with an ethanol extract of Zingiber officinale (400 mg/kg) extract along with a high-fat diet for 6 weeks. The extract of Zingiber officinale significantly decreased hepatic triglyceride and tended to decrease hepatic cholesterol levels when administered over 6 weeks to the rats fed a high-fat diet. We found that in parallel, the extract up-regulated both LDL receptor mRNA and protein level and down-regulated HMG-CoA reductase protein expression in the liver of these rats. The metabolic control of body lipid homeostasis is in part due to enhanced cholesterol biosynthesis and reduced expression of LDL receptor sites following long-term consumption of high-fat diets. The present results show restoration of transcriptional and post-transcriptional changes in low-density lipoprotein and HMG CoA reductase by Zingiber officinale administration with a high-fat diet and provide a rational explanation for the effect of ginger in the treatment of hyperlipidaemia.
Asunto(s)
Hidroximetilglutaril-CoA Reductasas/metabolismo , Hígado/efectos de los fármacos , Receptores de LDL/metabolismo , Zingiber officinale/química , Animales , Colesterol/metabolismo , Dieta , Grasas de la Dieta/administración & dosificación , Etanol/química , Grasas/metabolismo , Hidroximetilglutaril-CoA Reductasas/biosíntesis , Hidroximetilglutaril-CoA Reductasas/genética , Lipoproteínas LDL/metabolismo , Hígado/enzimología , Hígado/metabolismo , Extractos Vegetales/química , Extractos Vegetales/farmacología , ARN Mensajero/metabolismo , Ratas , Receptores de LDL/biosíntesis , Receptores de LDL/genética , Triglicéridos/metabolismoRESUMEN
Cajanus cajan L. is a natural plant, which contains a lot of potential active components. In the present study, we identified the effects of the stilbene extract from Cajanus cajan L. (sECC) on hepatic cholesterol metabolism in diet-induced (for 4 weeks) hyperlipidemic Kunming mice. All experimental mice were divided into 5 groups: control group, high lipid model group, sECC-treated with 200 or 100 mg kg(-1), and simvastatin (Sim, 12 mg kg(-1)) treated group. The mice were fed with fat and cholesterol-enriched chow except control mice that were fed with standard diet. The effects of sECC were investigated by monitoring serum and liver lipid profile (i. e. cholesterol homeostasis) in mice. To further explore the mechanism of sECC, hepatic cholesterol 7alpha-hydroxylase (CYP7A1) and low density lipoprotein (LDL) receptor expressions in cholesterol homeostasis were analyzed by reverse transcription PCR. After 4 weeks pretreatment, the mice in the high lipid model group showed markedly higher serum and hepatic lipid contents than control group (P< 0.01). Compared with high lipid model group, the increased serum and hepatic lipid contents were markedly attenuated by sECC (200 mg kg(-1)), the serum and hepatic total cholesterol were reduced by 31.5% and 22.7% (P<0.05), respectively. The triglyceride contents of serum and liver were also lowered by 23.0% and 14.4%, respectively. At the same times, serum LDL cholesterol decreased by 53.0% (P<0.01). The mRNA expressions of hepatic CYP7A1 and LDL-receptor were significantly enhanced in the mice administered with sECC (200 mg kg(-1)), whereas those expressions were suppressed by the fat and cholesterol-enriched diet. These data indicate that sECC reduces the atherogenic properties of dietary cholesterol in mice. It is indicated that expression enhancement of hepatic LDL-receptor and cholesterol 7alpha-hydroxylase may be responsible for the hypercholesterolemic effect.
Asunto(s)
Anticolesterolemiantes/farmacología , Colesterol 7-alfa-Hidroxilasa/biosíntesis , Hipercolesterolemia/metabolismo , Receptores de LDL/biosíntesis , Estilbenos/farmacología , Animales , Anticolesterolemiantes/aislamiento & purificación , Peso Corporal/efectos de los fármacos , Cajanus/química , Colesterol/sangre , Colesterol/metabolismo , Colesterol 7-alfa-Hidroxilasa/genética , LDL-Colesterol/sangre , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/farmacología , Regulación de la Expresión Génica , Hipercolesterolemia/sangre , Hipercolesterolemia/genética , Hipercolesterolemia/patología , Hígado/metabolismo , Hígado/patología , Masculino , Ratones , Tamaño de los Órganos/efectos de los fármacos , Hojas de la Planta/química , Plantas Medicinales/química , ARN Mensajero/metabolismo , Receptores de LDL/genética , Estilbenos/aislamiento & purificación , Triglicéridos/sangre , Triglicéridos/metabolismoRESUMEN
OBJECTIVE: To study lipid-regulating action of 2, 3, 5, 4'-tetrahydroxy stilbene-2-O-beta-D-glucopyranoside (TSG) from Polygonum multiflorum on experimental model hyperlipidemic rats. METHOD: TSG 90 and 180 mg x kg(-10 x d(-1), atorvastatin mg kg(-1) x d(-1) and saline 2 mL x d(-1) were administered to hyperlipidemic rats. Groups of rats were determined and compared with those of saline group. The LDLR and HMGR mRNA expression were also detected. RESULT: TSG significantly reduced serum TC and LDL-C level and atherosclerosis index, increased the expression of LDLR in the liver cells. CONCLUSION: TSG, which shows effects and mechanism in part like atorcastatin, is a major constituent with blood-lipid regulating effect of P. multiflorum and can be explored as a potent medication for hyperlipidemia. Effects on LDL-C and AI, as well as on gene expression of TSG were first reported.
Asunto(s)
Glucósidos/farmacología , Hiperlipidemias/sangre , Polygonum/química , Estilbenos/farmacología , Animales , Anticolesterolemiantes/administración & dosificación , Anticolesterolemiantes/farmacología , Atorvastatina , LDL-Colesterol/sangre , Glucósidos/administración & dosificación , Glucósidos/aislamiento & purificación , Hepatocitos/citología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Ácidos Heptanoicos/administración & dosificación , Ácidos Heptanoicos/farmacología , Hidroximetilglutaril-CoA Reductasas/biosíntesis , Hidroximetilglutaril-CoA Reductasas/genética , Hiperlipidemias/prevención & control , Masculino , Tubérculos de la Planta/química , Plantas Medicinales/química , Pirroles/administración & dosificación , Pirroles/farmacología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptores de LDL/biosíntesis , Receptores de LDL/genética , Estilbenos/administración & dosificación , Estilbenos/aislamiento & purificación , Triglicéridos/sangreRESUMEN
OBJECTIVE: To investigate the effect of reinioside C (RC) on the expression of lectin-like oxidized low density lipoprotein receptor (LOX)-1 mRNA and LOX-1 protein induced by oxidized low density lipoprotein (ox-LDL) in cultured human umbilical vein endothelial cells (HUVEC). METHODS: HUVECs were cultured with ox-LDL (50 mg/L) for 24 h in the absence or presence of RC (1, 3, and 10 micromol/L). The expressions of LOX-1 mRNA and LOX-1 protein were examined by RT-PCR and Western-blot. RESULTS: Incubation with ox-LDL (50 mg/L) significantly raised the expression of LOX-1 mRNA and LOX-1 protein,which was concentration-dependent. CONCLUSION: RC can inhibit the increased expression of LOX-1 mRNA and LOX-1 protein induced by ox-LDL in HUVECs.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Lipoproteínas LDL/farmacología , Polygala/química , Receptores de LDL/biosíntesis , Saponinas/farmacología , Venas Umbilicales/metabolismo , Células Cultivadas , Endotelio Vascular , Humanos , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Receptores de LDL/genética , Venas Umbilicales/citologíaRESUMEN
Conjugated linoleic acids (CLA) have attracted scientific interest due to their potential beneficial effects on atherosclerosis. Recently, a mixture of CLA isomers was demonstrated to upregulate LDL receptor expression in the human hepatoma cell line HepG2. However, the underlying mechanisms remain to be resolved. Thus, the aim of this study was to elucidate how CLA mediates upregulation of LDL receptor in HepG2 cells and whether this upregulation is isomer-specific. The results revealed that LDL receptor promoter activity and mRNA expression were strongly induced upon treatment with t10c12-CLA (P<0.05), whereas c9t11-CLA and linoleic acid (LA) had no effect. In addition, only treatment with t10c12-CLA markedly induced mRNA expression of SREBP-2 and HMG-CoA reductase and slightly induced that of SREBP-1 (P<0.05). Using SREBP-2 knockdown cells, we could demonstrate that the effect of t10c12-CLA on LDL receptor gene transcription was significantly reduced when compared to control cells (P<0.05). When using SREBP-1 knockdown cells the effect of t10c12-CLA on LDL receptor mRNA only slightly decreased compared to control cells. In addition, using different deletion constructs of the LDL receptor gene promoter we showed that the induction of the LDL receptor by t10c12-CLA is independent of the AP-1 motif in the LDL receptor promoter. In conclusion, the present study revealed that transcriptional activation of the LDL receptor gene by t10c12-CLA is dependent on the upregulation of SREBP-2 and is probably due to the activation of the SRE-1 in the LDL receptor gene promoter in HepG2 cells. Thus, the decreased plasma cholesterol levels in response to CLA as observed in a limited number of animal and human studies might be explained by an enhanced uptake of VLDL and LDL cholesterol via hepatic LDL receptors. However, it provides no explanation for the outcome of most human studies reporting unaltered or even increased plasma and LDL cholesterol concentrations in response to supplementation with CLA.
Asunto(s)
Ácidos Linoleicos Conjugados/farmacología , Receptores de LDL/genética , Línea Celular Tumoral , Colesterol/biosíntesis , Humanos , Hidroximetilglutaril-CoA Reductasas/genética , Ácido Linoleico/farmacología , Regiones Promotoras Genéticas/efectos de los fármacos , ARN Mensajero/metabolismo , Receptores de LDL/biosíntesis , Transducción de Señal/efectos de los fármacos , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/genética , Proteína 2 de Unión a Elementos Reguladores de Esteroles/genética , Factor de Transcripción AP-1/metabolismo , Regulación hacia ArribaRESUMEN
Casein-based diets containing a low (LDI) or high (HDI) dose of soya protein concentrate enriched with isoflavones were fed to obese Zucker rats for 6 weeks. HDI feeding, but not LDI feeding, reduced the fatty liver and decreased the plasma levels of alanine transaminase and aspartate transaminase. This was accompanied by increased activities of mitochondrial and peroxisomal beta-oxidation, acetyl-CoA carboxylase, fatty acid synthase and glycerol-3-phosphate acyltransferase in liver and increased triacylglycerol level in plasma. The decreased fatty liver and the increased plasma triacylglycerol level appeared not to be caused by an increased secretion of VLDL, as HDI decreased the hepatic mRNA levels of apo B and arylacetamide deacetylase. However, the gene expression of VLDL receptor was markedly decreased in liver, but unchanged in epididymal white adipose tissue and skeletal muscle of rats fed HDI, indicating that the liver may be the key organ for the reduced clearance of triacylglycerol-rich lipoproteins from plasma after HDI feeding. The n-3/n-6, 20:4n-6/18:2n-6 and (20:5n-3+22:6n-3)/18:3n-3 ratios were increased in liver triacylglycerol by HDI. The phospholipids in liver of rats fed HDI contained a low level of 20:4n-6 and a high level of 20:5n-3, favouring the production of anti-inflammatory eicosanoids. When obese Zucker rats were fed soya protein, this also resulted in reduced fatty liver, possibly through reduced clearance of VLDL by the liver. We conclude that the isoflavone-enriched soya concentrate as well as soya protein may be promising dietary supplements for treatment of non-alcoholic fatty liver.
Asunto(s)
Suplementos Dietéticos , Hígado Graso/dietoterapia , Isoflavonas/uso terapéutico , Hígado/metabolismo , Proteínas de Soja/uso terapéutico , Animales , Ácidos Grasos/metabolismo , Hígado Graso/etiología , Hígado Graso/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Metabolismo de los Lípidos/efectos de los fármacos , Masculino , Obesidad/complicaciones , Obesidad/metabolismo , Oxidación-Reducción , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Zucker , Receptores de LDL/biosíntesis , Receptores de LDL/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
OBJECTIVE: To investigate the effect of soybean isoflavone on blood lipid and the expression of LDLR mRNA in ovariectomied rats. METHODS: Forty eight Sprague-Dawley rats were divided randomly into four groups: Sham group (sham), OVX group (OVX), OVX + gemifibrozil (G) group (OVX + G) and OVX + isoflavone (ISO) group (OVX_ISO). in which the rats were treated with G or ISO for three months starting from two weeks after both sides of rat' s ovarietomy. Blood sample were taken out for determination of blood lipid. The liver tissue were taken out quickly, Isothiocyanate guanidine-phenol-chloroform was used to extract the total RNA from the liver and RT-PCR was used to detect the expression of LDLR mRNA. RESULTS: Compared with the OVX group, contents of TG, LDL-C and OX-LDL in serum in OVX + ISO group decreased remarkably, and the contents of HDL-C increased. LDLR mRNA in OVX + ISO group increased distinctively compared with OVX group and OVX + G group. CONCLUSION: The level of mRNA of LDLR in liver decreas after ovariotormy and the soybean isoflavone may increase it by regulating the expression level of LDLR through transcription.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Glycine max/química , Isoflavonas/farmacología , Hígado/efectos de los fármacos , Receptores de LDL/biosíntesis , Animales , Arteriosclerosis/prevención & control , HDL-Colesterol/sangre , Femenino , Hígado/metabolismo , Ovariectomía , ARN Mensajero/biosíntesis , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptores de LDL/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Triglicéridos/sangreRESUMEN
Selenium deficiency has been associated with hypercholesterolemia. Present study was aimed to determine the effect of selenium (Se) deficiency on LDL receptor (LDL-R) activity as well as mRNA expression during experimental hypercholesterolemia in SD male rats. Animals were fed Se adequate (0.2 ppm) and deficient (0.02 ppm) control diet as well as high cholesterol (2%) diet (HCD) for 1 and 2 months. LDL-R activity was measured in vivo by injecting radiolabeled LDL to rats and percent decrease in cpm with time was taken as a measure of LDL clearance and in turn LDL-R activity. LDL-R mRNA expression was studied by RT-PCR. LDL-R activity and mRNA expression decreased significantly on HCD feeding in both Se deficient and adequate diet fed rats after 2 months. In Se deficiency receptor activity and mRNA expression decreased significantly. After 2 months LDL-R activity and expression decreased in both the Se deficient groups and in Se adequate HCD fed group in comparison to 1 month data. But after 4 month there was no significant difference observed in LDL-R activity and mRNA expression in selenium deficiency as well as on HCD feeding. So the present results demonstrate that Se deficiency act synergistically with hypercholesterolemia to downregulate LDL-R activity as well as mRNA expression.
Asunto(s)
Hipercolesterolemia/metabolismo , Receptores de LDL/biosíntesis , Selenio/deficiencia , Animales , Colesterol/sangre , Colesterol en la Dieta/administración & dosificación , Regulación hacia Abajo , Glutatión Peroxidasa/metabolismo , Hipercolesterolemia/etiología , Lipoproteínas LDL/sangre , Hígado/metabolismo , Masculino , ARN Mensajero/biosíntesis , Ratas , Ratas Sprague-Dawley , Receptores de LDL/genética , Triglicéridos/sangreRESUMEN
BACKGROUND: Apolipoprotein B (apoB) contains ligand-binding domain for the binding of LDL to LDL-R site, which enables the removal of LDL from circulation. Our recent data showed that selenium (Se) is involved in the lipid metabolism. The present study was aimed to understand the effect of Se deficiency (0.02 ppm) and selenium supplementation (1 ppm) on apoB expression in liver during hypercholesterolemia in male Sprague Dawley rats. Animals were fed with control and high cholesterol diet (2%) for 1 and 2 months. ApoB levels by ELISA and protein expression by western blot was done. Hepatic LDL receptor (LDL-R) activity (in vivo) and mRNA expression by RT-PCR was monitored. RESULTS: In selenium deficiency and on high cholesterol diet (HCD) feeding apoB levels increased and LDL-R expression decreased significantly after 2 months. On 1 ppm selenium supplementation apoB expression significantly decreased and LDL-R expression increased after 2 months. But after one month of treatment there was no significant change observed in apoB and LDL-R expression. CONCLUSION: So the present study demonstrates that Se deficiency leads to up regulation of apoB expression during experimental hypercholesterolemia. Selenium supplementation upto 1 ppm leads to downregulation of apoB expression. Further, this study will highlight the nutritional value of Se supplementation in lipid metabolism.
Asunto(s)
Apolipoproteínas B/biosíntesis , Hipercolesterolemia/metabolismo , Selenio/administración & dosificación , Selenio/deficiencia , Animales , Colesterol/sangre , Colesterol en la Dieta/administración & dosificación , LDL-Colesterol/sangre , Regulación hacia Abajo/efectos de los fármacos , Glutatión Peroxidasa/metabolismo , Yoduro Peroxidasa/metabolismo , Masculino , Ratas , Ratas Sprague-Dawley , Receptores de LDL/biosíntesis , Selenio/sangre , Tiroxina/sangre , Triyodotironina/sangre , Regulación hacia ArribaRESUMEN
OBJECTIVE: To investigate the molecular mechanism of curcumin in reducing blood lipids by establishing gene expression system of human low density lipoprotein receptors (LDL-R) in Xenopus Laevis oocytes (XLO). METHODS: The expression of LDL-R on cytomembrane was determined using immuno-fluorescent, ligand-fluorescent and immune colloidal gold techniques after human LDL-R containing p3.7 LDL plasmid was led into nucleus. And the expression of LDL-R gene in XLO was quantitatively determined by ELISA after being interfered with different concentrations of curcumin. RESULTS: The human LDL-R gene could be expressed on XLO, which could be significantly enhanced by curcumin in a dose-dependent manner. Conclusion One of the paths of curcumin in reducing blood lipids and anti-atherosclerosis was improving LDL-R gene expression and increasing the LDL-cholesterol absorption of cells.
Asunto(s)
Curcumina/farmacología , Oocitos/metabolismo , Receptores de LDL/biosíntesis , Animales , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Regulación de la Expresión Génica , Humanos , Microinyecciones , Oocitos/citología , Receptores de LDL/genética , Xenopus laevisRESUMEN
OBJECTIVE: To elucidate the molecular mechanism of Wendan Tang in prevention of lipid metabolism disorder in adult rats. METHOD: On the basis of hyperlipidemia rat models, triglycerides (TG), total cholesterol (TC) in serum, activities of lipase (LA), lipoprotein lipase (LPL), hepatic lipase (HL) in liver, parts of hemogram and hepatic LDLR mRNA levels were investigated 21 days after the feeding of atherogenic diet. RESULT: Wendan Tang significantly reduced the serum TG, TC and increased the activity of LPL and LA, but caused no chang in HL. The result of RT-PCR test showed that high fat and high cholesterol feeding could significantly induce the reduction of LDLR mRNA levels, while Wendan Tang could increase hepatic LDLR density. CONCLUSION: Wendan Tang can prevent disorder of lipid metabolism by regulating TC, TG, LDL-c through upregaulation of LDLR transcription level and improving antioxidant ability.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Hiperlipidemias/sangre , Metabolismo de los Lípidos , Receptores de LDL/biosíntesis , Animales , Combinación de Medicamentos , Medicamentos Herbarios Chinos/aislamiento & purificación , Femenino , Hiperlipidemias/metabolismo , Hiperlipidemias/prevención & control , Hígado/metabolismo , Masculino , Plantas Medicinales/química , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Ratas , Receptores de LDL/genéticaRESUMEN
OBJECTIVE: To study the influence of Jiangzhining decoction on the genetic expression of Liver LDLR of the rats suffered from hyperlipemia. METHOD: Laboratory animals were male wister rats with hyperlipemia resulting from high fat feeding. Prescription was the douche of stomach with Jiangzhining decoction (200%) with a dosage of 1.4 g.kg-1, for 15 successive days. Total RNA was extracted from the liver tissue of treated rats and LDLRmRNA was detected by Dot blot hybridization. Expression levels of LDLRmRNA was estimated by a ratio of LDLRmRNA and beta-actin mRNA. RESULT: The difference between expression levels of LDLRmRNA for normal group and those for hyperlipemia group (100% +/- 19% vs 39% +/- 14%) was significant (P < 0.05); and the difference between decoction group (108 +/- 8%) and hyperlipimia group was also highly significant (P < 0.01). CONCLUSION: High fat feeding reduces the expression of liver LDLRmRNA while the decoction can greatly increase it. The study and development of Jiangzhining are significant in preventing and curing cadiocerebral diseases.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Hiperglucemia/metabolismo , Hipoglucemiantes/farmacología , Hígado/metabolismo , Receptores de LDL/biosíntesis , Actinas/biosíntesis , Actinas/genética , Animales , Combinación de Medicamentos , Medicamentos Herbarios Chinos/administración & dosificación , Hipoglucemiantes/administración & dosificación , Masculino , ARN Mensajero/genética , Ratas , Ratas Wistar , Receptores de LDL/genéticaRESUMEN
The action of LY295427 [(3alpha,4alpha, 5alpha)-4-(2-propenylcholestan-3-ol)], a compound that derepresses low-density lipoprotein receptor (LDL-R) expression in a cell-based model, was examined in hamsters. It was found that the compound does not have an effect in normal chow-fed hamsters, in which LDL-R levels are not repressed, but exerts a marked hypocholesterolemic effect (>70% decrease) in cholesterol-coconut oil-fed hamsters, in which LDL-R is repressed. In this model, there is a dose-response for cholesterol lowering with an approximate ED50 value of 40 mg/kg/day and an inverse relationship between serum cholesterol and serum LY295427 levels. LDL-R mRNA is increased (2-fold) and liver cholesterol ester content is decreased (>90%). Unlike the 3-hydroxy-3-methylglutarylcoenzyme A reductase inhibitor lovastatin, the decreased serum cholesterol is confined to the non-high-density lipoprotein fraction. Furthermore, LY295427 does not affect cholesterol biosynthesis, and it does not have a significant effect on cholesterol absorption. These data suggest that LY295427 acts in the hypercholesterolemic hamster by derepressing LDL-R transcription, thereby enhancing cholesterol clearance from the blood. The results with LY295427 suggest that compounds that act to increase LDL-R may represent a novel approach in the pharmacotherapy for hypercholesterolemia.
Asunto(s)
Anticolesterolemiantes/farmacología , Colestanoles/farmacología , Colesterol/metabolismo , Hipercolesterolemia/metabolismo , Receptores de LDL/genética , Regulación hacia Arriba/efectos de los fármacos , Acetatos/metabolismo , Animales , Colesterol/sangre , Colesterol en la Dieta/administración & dosificación , LDL-Colesterol/sangre , Aceite de Coco , Cocos/química , Cricetinae , Grasas de la Dieta/administración & dosificación , Homeostasis/efectos de los fármacos , Absorción Intestinal/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Lovastatina/farmacología , Masculino , Mesocricetus , Aceites de Plantas/administración & dosificación , ARN Mensajero/biosíntesis , Receptores de LDL/biosíntesisRESUMEN
Consumption of boiled coffee raises serum cholesterol levels in humans. The diterpenes cafestol and kahweol in boiled coffee have been found to be responsible for the increase. To investigate the biochemical background of this effect, we studied the effects of cafestol and a mixture of cafestol/kahweol/isokahweol (48:47:5 w/w) on bile acid synthesis and cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase in cultured rat hepatocytes. Dose-dependent decreases of bile acid mass production and cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase activity were found, showing a maximal reduction of -91%, -79%, and -49% respectively, at a concentration of 20 micrograms/mL cafestol. The decrease in 7 alpha-hydroxylase and 27-hydroxylase activity paralleled well the suppression of the respective mRNAs, being -79% and -77%, and -49% and -46%, respectively, at 20 micrograms/mL cafestol. Run-on data showed a reduction in 7 alpha-hydroxylase and 27-hydroxylase gene transcriptional activity after incubation with cafestol. The mixture of cafestol/kahweol/isokahweol was less potent in suppression of bile acid synthesis and cholesterol 7 alpha-hydroxylase. Cafestol (20 micrograms/mL) had no effect on lithocholic acid 6 beta-hydroxylase mRNA, another enzyme involved in bile acid synthesis. LDL-receptor, HMG-CoA reductase, and HMG-CoA synthase mRNAs were significantly decreased by cafestol (-18%, -20%, and -43%, respectively). We conclude that cafestol suppresses bile acid synthesis by downregulation of cholesterol 7 alpha-hydroxylase and of, to a lesser extent, sterol 27-hydroxylase in cultured rat hepatocytes, whereas kahweol and isokahweol are less active. We suggest that suppression of bile acid synthesis may provide an explanation for the cholesterol-raising effect of cafestol in humans.
Asunto(s)
Ácidos y Sales Biliares/biosíntesis , Colesterol 7-alfa-Hidroxilasa/antagonistas & inhibidores , Café/química , Inhibidores Enzimáticos del Citocromo P-450 , Diterpenos/farmacología , Hipercolesterolemia/inducido químicamente , Hígado/efectos de los fármacos , Esteroide Hidroxilasas/antagonistas & inhibidores , Animales , Colestanotriol 26-Monooxigenasa , Colesterol/metabolismo , Colesterol 7-alfa-Hidroxilasa/genética , Colesterol 7-alfa-Hidroxilasa/metabolismo , Ésteres del Colesterol/metabolismo , Café/efectos adversos , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Diterpenos/aislamiento & purificación , Inducción Enzimática/efectos de los fármacos , Calor , Hidroximetilglutaril-CoA Reductasas/biosíntesis , Hidroximetilglutaril-CoA Reductasas/genética , Hidroximetilglutaril-CoA Sintasa/biosíntesis , Hidroximetilglutaril-CoA Sintasa/genética , Hígado/enzimología , Masculino , ARN Mensajero/biosíntesis , Ratas , Ratas Wistar , Receptores de LDL/biosíntesis , Receptores de LDL/genética , Esteroide Hidroxilasas/genética , Esteroide Hidroxilasas/metabolismo , Terpenos/farmacología , Transcripción Genética/efectos de los fármacos , Triglicéridos/metabolismoRESUMEN
Potocytosis is an endocytic process that is specialized for the internalization of small molecules. Recent studies on the uptake of 5-methyltetrahydrofolate by the folate receptor have suggested that the glycosyl-phosphatidylinositol anchor on this protein causes it to cluster and be internalized by caveolae instead of coated pits. To test this hypothesis directly, we have constructed a chimeric folate receptor that has the glycosyl-phosphatidylinositol anchor replaced with the transmembrane domain and cytoplasmic tail of the low density lipoprotein receptor. The cells with wild-type receptors delivered 5-methyltetrahydrofolate to the cytoplasm more rapidly than did cells expressing the chimeric receptor. This suggests that efficient delivery to the cytoplasm depends on caveolae. In sharp contrast to cells with wild-type folate receptors, cells internalizing folate by clathrin-coated pits were unable to decrease vitamin uptake when they were either folate replete or confluent.
Asunto(s)
Proteínas Portadoras/metabolismo , Clatrina/metabolismo , Invaginaciones Cubiertas de la Membrana Celular/metabolismo , Endocitosis , Receptores de Superficie Celular , Receptores de LDL/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Tetrahidrofolatos/metabolismo , Animales , Secuencia de Bases , Células CHO , Proteínas Portadoras/biosíntesis , División Celular , Cricetinae , Citoplasma/metabolismo , Cartilla de ADN , ADN Complementario , Técnica del Anticuerpo Fluorescente , Receptores de Folato Anclados a GPI , Glicosilfosfatidilinositoles/metabolismo , Humanos , Cinética , Datos de Secuencia Molecular , Plásmidos , Receptores de LDL/biosíntesis , Proteínas Recombinantes de Fusión/biosíntesis , TransfecciónRESUMEN
The concentration of LDL in plasma is strongly influenced by the amount and the type of lipid in the diet. Recent studies in the hamster have shown that dietary fatty acids differentially affect circulating LDL levels primarily by altering receptor-dependent LDL uptake in the liver. To investigate the mechanistic basis of this effect, rates of receptor-dependent LDL transport in the liver were correlated with LDL receptor protein and mRNA levels in hamsters fed safflower oil or coconut oil and varying amounts of cholesterol. Hepatic LDL receptor activity was significantly lower in animals fed coconut oil than in animals fed safflower oil at all levels of cholesterol intake (26, 53, and 61% lower at cholesterol intakes of 0, 0.06, and 0.12%, respectively). These fatty acid-induced changes in hepatic LDL receptor activity were accompanied by parallel changes in hepatic LDL receptor protein and mRNA levels, suggesting that dietary fatty acids regulate the LDL receptor pathway largely at the mRNA level.