Expression Of Very Low-density Lipoprotein Receptor Research Articles

Silencing Very-Low-Density Lipoprotein Receptor Reveals Alpha-1 Antitrypsin Role in HIV Infectivity.

Here we describe methods for investigating alpha-1 antitrypsin (AAT) and very-low-density lipoprotein receptor (VLDLR) interactions with infectious and non-infectious HIV-1 virions. Using silencing RNA to transiently block expression of VLDLR and the receptor-associated protein (RAP) to continuously block VLDLR activity, AAT is demonstrated to participate with VLDLR during internalization and infectivity of HIV-1 virions.

Expression level of VLDL receptor and VLDL-c levels in the malignant and benign breast tumors: The correlation with miRNA-4465 and miRNA-1297

Breast cancer as one of the most prevalent cancers has high morbidity and mortality. Very low-density lipoprotein receptor (VLDLR) is a multifunctional receptor which plays a principal role in the tumor development through affecting cell metastasis and proliferation. The VLDLR as a target for miRNA-4465 and miRNA-1297 was predicted using bioinformatics analysis. Tissue specimens of malignant (n = 50), benign (n = 35) and corresponding normal breast (n = 20) were considered to evaluate the expression of VLDLR using RT-qPCR and western blotting. The VLDL cholesterol (VLDL-C) levels were quantified using a colorimetric assay. The relative VLDLR expression was found in the malignant tumors, which was significantly lower than that in the normal tissues (P<0.05). The expression levels of VLDLR had no significant difference between malignant and benign tissues (P>0.05). Correlation analysis revealed that the VLDLR expression level had a direct correlation with miRNA-1297 (R=0.566, P<0.05), but a reverse one with miRNA-4465 (R = −0.663, P<0.0001). The VLDL-C level in the malignant and normal tissues was lower than that in the benign tumors, which was not significant (P>0.05). The expression levels of VLDLR in E+P–H- (ER+,PR−,HER2-) tumors were higher than those in other subtypes (P<0.05). Furthermore, a negative correlation was found between the VLDLR expression level and the Ki 67% score. These data revealed that the lower expression of VLDLR mediated by the high expression levels of miRNA-4465 may be involved in the development of breast cancer. These results might provide some evidence for the effect of VLDLR on the breast cancer.

Pharmacological PPARβ/δ activation upregulates VLDLR in hepatocytes

The very low-density lipoprotein receptor (VLDLR) plays an important function in the control of serum triglycerides and in the development of non-alcoholic fatty liver disease (NAFLD). In this study, we investigated the role of peroxisome proliferator-activated receptor (PPAR)β/δ activation in hepatic VLDLR regulation. Treatment of mice fed a high-fat diet with the PPARβ/δ agonist GW501516 increased the hepatic expression of Vldlr. Similarly, exposure of human Huh-7 hepatocytes to GW501516 increased the expression of VLDLR and triglyceride accumulation, the latter being prevented by VLDLR knockdown. Finally, treatment with another PPARβ/δ agonist increased VLDLR levels in the liver of wild-type mice, but not PPARβ/δ-deficient mice, confirming the regulation of hepatic VLDLR by this nuclear receptor. Our results suggest that upregulation of hepatic VLDLR by PPARβ/δ agonists might contribute to the hypolipidemic effect of these drugs by increasing lipoprotein delivery to the liver. Overall, these findings provide new effects by which PPARβ/δ regulate VLDLR levels and may influence serum triglyceride levels and NAFLD development.

Pharmacological PPARβ/δ activation upregulates VLDLR in hepatocytes

The very low-density lipoprotein receptor (VLDLR) plays an important function in the control of serum triglycerides and in the development of non-alcoholic fatty liver disease (NAFLD). In this study, we investigated the role of peroxisome proliferator-activated receptor (PPAR)β/δ activation in hepatic VLDLR regulation. Treatment of mice fed a high-fat diet with the PPARβ/δ agonist GW501516 increased the hepatic expression of Vldlr. Similarly, exposure of human Huh-7 hepatocytes to GW501516 increased the expression of VLDLR and triglyceride accumulation, the latter being prevented by VLDLR knockdown. Finally, treatment with another PPARβ/δ agonist increased VLDLR levels in the liver of wild-type mice, but not PPARβ/δ-deficient mice, confirming the regulation of hepatic VLDLR by this nuclear receptor. Our results suggest that upregulation of hepatic VLDLR by PPARβ/δ agonists might contribute to the hypolipidemic effect of these drugs by increasing lipoprotein delivery to the liver. Overall, these findings provide new effects by which PPARβ/δ regulate VLDLR levels and may influence serum triglyceride levels and NAFLD development.

Roles of Adipose Tissue VLDL-VLDLR Axis in Energy Metabolism

Elevated plasma triglyceride-rich lipoprotein, such as very low-density lipoprotein (VLDL), is considered a risk factor for prevalence of obesity, type 2 diabetes mellitus, and atherosclerosis. VLDL receptor (VLDLR), one of the lipoprotein receptor family proteins, is involved in clearance of circulating VLDL. In obese animals, adipose tissue macrophages (ATMs) contain high level of lipid metabolites, which seems to be linked to pro-inflammatory responses. However, it has not been thoroughly understood whether lipid uptake via VLDLR in macrophages is involved in obesity-induced inflammatory responses and insulin resistance. In this study, we demonstrate that elevated VLDLR in ATMs could accelerate adipose tissue inflammation and glucose intolerance in obesity. Expression of VLDLR was increased in ATMs from obese mice. In VLDL-treated macrophages, the level of ceramides was elevated through VLDLR, which potentiates pro-inflammatory responses and augments M1-like macrophage polarization. Moreover, adoptive transfer of VLDLR knockout bone marrow to wild type mice relieved adipose tissue inflammation and improved insulin resistance in diet-induced obesity. Taken together, these findings suggest that increased VLDL-VLDLR axis in ATMs would aggravate adipose tissue inflammation and insulin resistance in obesity. Disclosure J. Kim: None. K. Shin: None. I. Hwang: None. S. Choe: None.

Decreased expression of VLDLR is inversely correlated with miR-200c in human colorectal cancer.

Colorectal cancer (CRC) is one of the most common cancers and has a high rate of morbidity and mortality worldwide. Very-low-density-lipoprotein receptor (VLDLR), a member of the low-density-lipoprotein receptor (LDLR) superfamily, is a multifunctional receptor that regulates cellular signaling by binding numerous ligands. Several studies reported the altered expression of VLDLR and suggested that VLDLR may play a critical role in tumor development by affecting cell proliferation and metastasis. However, the function of VLDLR and regulation of its expression by miRNAs have not been investigated in CRC. In the present study, we investigated the expression of VLDLR in CRC patients and found it to be significantly decreased in tumors in comparison with paired adjacent non-tumor tissues. Moreover, VLDLR over-expression inhibited the proliferation and migration of CRC cells. We also found that VLDLR expression was negatively regulated by miR-200c in CRC cells and that their expression levels were inversely correlated in CRC patients. These data suggest that VLDLR down-regulation mediated by the increased expression of miR-200c may be involved in the development of CRC.

Abstract 411: Obesity Alters the Expression of Very Low Density Lipoprotein Receptor (vldlr) Expression in Peripheral Blood Monocytes: Possible Links Between Vldlr Expression and Monocyte Phenotypes

Objective: Very low density lipoprotein receptor (VLDLR) is a member of the low-density lipoprotein receptor family that binds multiple ligands including TG-rich lipoproteins. The role of VLDLR in lipid uptake had been demonstrated in cell culture and animal models, but little is known about the effects of obesity on VLDLR expression in human. Accordingly, we examined the impact of obesity on VLDLR expression in peripheral blood monocytes and explored possible links with monocyte inflammatory markers and circulating lipids. Subjects and measurements: Blood was collected from a total of 69 subjects including 23 normal-weight (body mass index: BMI &lt; 25 kg/m 2 ) controls and 46 obese (BMI &gt; 30 kg/m 2 ) subjects. Blood lipids, glucose and hormones were measured. Monocytes were isolated from buffy coats using Ficoll density centrifugation followed by magnetic separation of CD14-positive cells with magnetic beads. Expression of VLDLR, and pro- and ant-inflammatory markers of monocytes was examined by western blotting and quantitative qPCR. The uptake of labeled VLDL (DiI-VLDL) was also assessed in monocyte cultures. Results: Insulin, and total and VLDL triglycerides were increased in plasma of obese subjects. In addition, VLDLR protein and mRNA levels were significantly (p&lt; 0.05) higher (~ 40%) in obese compared to lean subjects. Monocytes of obese subjects exhibited higher expression of IL-6 and TNF-α (+29 to 43%), but lower expression of IL-10 and CD163 (-30 to 47%). The levels of monocyte VLDLR was positively correlated with blood triglycerides and insulin, and negatively with adiponectin. The uptake of labeled VLDL, assessed by fluorescence microscope, was higher in monocytes of obese subjects. Conclusions: Our findings indicate that obesity upregulates the expression of VLDLR and pro-inflammatory markers in peripheral blood monocytes. The uptake VLDL was also increased in monocyte of obese subjects suggesting that obesity-induced VLDLR expression may explain the changes of VLDL uptake and monocyte phenotype.

Hepatitis C virus utilizes VLDLR as a novel entry pathway

Various host factors are involved in the cellular entry of hepatitis C virus (HCV). In addition to the factors previously reported, we discovered that the very-low-density lipoprotein receptor (VLDLR) mediates HCV entry independent of CD81. Culturing Huh7.5 cells under hypoxic conditions significantly increased HCV entry as a result of the expression of VLDLR, which was not expressed under normoxic conditions in this cell line. Ectopic VLDLR expression conferred susceptibility to HCV entry of CD81-deficient Huh7.5 cells. Additionally, VLDLR-mediated HCV entry was not affected by the knockdown of cellular factors known to act as HCV receptors or HCV entry factors. Because VLDLR is expressed in primary human hepatocytes, our results suggest that VLDLR functions in vivo as an HCV receptor independent of canonical CD81-mediated HCV entry.

Activation of PPAR-δ induces microRNA-100 and decreases the uptake of very low-density lipoprotein in endothelial cells.

Increased level of very low-density lipoprotein (VLDL) is a key feature of the metabolic syndrome and is associated with cardiovascular diseases. PPAR-δ agonists play a protective role in lipid metabolism and vascular function. In this study, we aimed to investigate the role of PPAR-δ in the uptake of VLDL in endothelial cells and its underlying mechanism(s). Uptake of VLDL in HUVECs was assessed by Dil-fluorescent labelling of VLDL. Levels of VLDL receptor mRNA and microRNA (miR-100) were detected by quantitative PCR. The target genes of miR-100 were predicted using bioinformatics analysis. 3'-Untranslated region (3'-UTR) luciferase reporter and Argonaute 1 pull-down assays were used to validate the target of miR-100. PPAR-δ agonist GW501516 decreased uptake of VLDL and expression of VLDL receptor at mRNA and protein levels. GW501516 inhibited the luciferase reporter activity of the 3'-UTR of VLDL receptor. VLDL receptor was a direct target of miR-100. miR-100 was significantly increased by GW501516 in HUVECs. Transfection of a miR-100 mimic decreased the mRNA and protein levels of VLDL receptor and uptake of VLDL. Furthermore, a miR-100 inhibitor abolished the inhibitory effect of PPAR-δ on VLDL receptor expression and VLDL uptake. In endothelial cells, activation of PPAR-δ decreased VLDL receptor expression and VLDL uptake via the induction of miR-100. These results provided a novel mechanism for the vascular protective effect of PPAR-δ agonists.

Endoplasmic reticulum stress induces hepatic steatosis via increased expression of the hepatic very low‐density lipoprotein receptor

Recent evidence suggests that obese animals exhibit increased endoplasmic reticulum (ER) stress in the liver and adipose tissue. Although ER stress is closely associated with lipid homeostasis, it is largely unknown how ER stress contributes to hepatic steatosis. In this study, we demonstrate that the induction of ER stress stimulates hepatic steatosis through increased expression of the hepatic very low-density lipoprotein receptor (VLDLR). Among the unfolded protein response sensors, the protein kinase RNA-like ER kinase-activating transcription factor 4 signaling pathway was required for hepatic VLDLR up-regulation. In primary hepatocytes, ER stress-dependent VLDLR expression induced intracellular triglyceride accumulation in the presence of very low-density lipoprotein. Moreover, ER stress-dependent hepatic steatosis was diminished in the livers of VLDLR-deficient and apolipoprotein E-deficient mice compared with wild-type mice. In addition, the VLDLR-deficient mice exhibited decreased hepatic steatosis upon high-fat diet feeding. These data suggest that ER stress-dependent expression of hepatic VLDLR leads to hepatic steatosis by increasing lipoprotein delivery to the liver, which might be a novel mechanism explaining ER stress-induced hepatic steatosis.

Low density lipoprotein receptor–related protein 1 is upregulated in epicardial fat from type 2 diabetes mellitus patients and correlates with glucose and triglyceride plasma levels

Lipoprotein receptor expression plays a crucial role in the pathophysiology of adipose tissue in in vivo models of diabetes. However, there are no studies in diabetic patients. The aims of this study were to analyze (a) low-density lipoprotein receptor-related protein 1 (LRP1) and very low-density lipoprotein receptor (VLDLR) expression in epicardial and subcutaneous fat from type 2 diabetes mellitus compared with nondiabetic patients and (b) the possible correlation between the expression of these receptors and plasmatic parameters. Adipose tissue biopsy samples were obtained from diabetic (n = 54) and nondiabetic patients (n = 22) undergoing cardiac surgery before the initiation of cardiopulmonary bypass. Adipose LRP1 and VLDLR expression was analyzed at mRNA level by real-time PCR and at protein level by Western blot analysis. Adipose samples were also subjected to lipid extraction, and fat cholesterol ester, triglyceride, and free cholesterol contents were analyzed by thin-layer chromatography. LRP1 expression was higher in epicardial fat from diabetic compared with nondiabetic patients (mRNA 17.63 ± 11.37 versus 7.01 ± 4.86; P = 0.02; protein 11.23 ± 7.23 versus 6.75 ± 5.02, P = 0.04). VLDLR expression was also higher in epicardial fat from diabetic patients but only at mRNA level (231.25 ± 207.57 versus 56.64 ± 45.64, P = 0.02). No differences were found in the expression of LRP1 or VLDLR in the subcutaneous fat from diabetic compared with nondiabetic patients. Epicardial LRP1 and VLDLR mRNA overexpression positively correlated with plasma triglyceride levels (R(2) = 0.50, P = 0.01 and R(2) = 0.44, P = 0.03, respectively) and epicardial LRP1 also correlated with plasma glucose levels (R(2) = 0.33, P = 0.03). These results suggest that epicardial overexpression of certain lipoprotein receptors such as LRP1 and VLDLR expression may play a key role in the alterations of lipid metabolism associated with type 2 diabetes mellitus.

P3-17-01: ApoE and Its Receptors (LRP8, VLDLR) Function as Growth Signals for Triple-Negative Breast Cancer and Represent a Novel Therapeutic Target.

Abstract Triple-negative breast cancers (TNBC) lack the expression of estrogen, progesterone, and the human epidermal growth factor 2 receptors and have limited treatment options. We hypothesized that genes that are frequently amplified or overexpressed in these cancers are functionally important for the growth and survival of triple-negative breast cancer. Materials/Methods: We identified genes overexpressed in TNBC in 3 different human breast cancer gene expression data sets (n=294, n=286, n=198). We assessed the functional importance of the consistently overexpressed genes using siRNA screen on 18 breast cancer cell lines (10 ER-, 8 ER+) in vitro. We targeted each gene with 4 different siRNA constructs separately and each experiment was performed in triplicate. The genes with the greatest TNBC-specific inhibitory effect after siRNA down regulation were selected for further mechanistic and signal transduction studies. Results: 684 genes showed consistent and highly significant overexpression in TNBC compared to receptor-positive cancers in all 3 data sets. siRNA suppression of 161 of these genes inhibited cell growth significantly more in the ER-negative compared to ER-positive cells by at least 1 of the 4 siRNAs, 27 genes showed similar effect with 2 or more siRNAs and for 2 genes 3 of the 4 siRNAs showed preferential growth inhibition in ER-negative cells. These two genes were VLDLR (very low-density lipoprotein receptor) and LRP8 (low density lipoprotein receptor-related protein 8). We validated the siRNA screen results and confirmed down regulation of the mRNA and protein levels for VLDLR and LRP8 in 4 different ER-negative cell lines and showed that siRNA inhibition can be rescued by co-transfection of the receptor genes. Reanalysis of gene expression data also indicated expression of the LRP8 and VLDLR ligands, Reelin or ApoE, in both breast cancer tissues and in cell lines. We next demonstrated that exposure to Reelin and ApoE stimulates the growth of ER-negative cells in vitro. The stimulatory effect of ApoE was isoform dependent ApoE2 (Cys112/Cys158;) has the lowest receptor binding affinity and showed no growth stimulation, ApoE3 (Cys112/ Arg158) had modest 50–60% growth stimulation and ApoE4 (Arg112/ Arg158) had the greatest stimulatory effect (300-400%). Suppression of the expression of either LRP8 or VLDLR or exposure to RAP (an inhibitor of ligand binding to LRP8 and VLDLR) abolished this ligand-induced proliferation. ApoE4 stimulation results in the transcriptional upregulation of genes involved in proliferation, metabolism, and inflammatory signaling pathways. ApoE4 stimulation increases expression of proteins involved in MAPK/ERK pathway, DNA damage repair, and inflammation. Conclusions: We show that theApoE - LRP8/VLDLR ligand receptor system is overexpressed in human TNBC. We also demonstrated that this receptor system mediates a strong growth promoting and survival function in TNBC cells in vitro. Interestingly, allelic imbalance favoring ApoE4 expression (E3/E4 or E4/E4) has been linked to higher risk of developing early onset breast cancer, which is primarily TNBC. We propose that inhibitors of LRP8/VLDLR signaling may be clinically useful therapeutic or preventive agents for TNBC. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-17-01.

Retinoic Acid-inducible Gene I-inducible miR-23b Inhibits Infections by Minor Group Rhinoviruses through Down-regulation of the Very Low Density Lipoprotein Receptor

In mammals, viral infections are detected by innate immune receptors, including Toll-like receptor and retinoic acid inducible gene I (RIG-I)-like receptor (RLR), which activate the type I interferon (IFN) system. IFN essentially activates genes encoding antiviral proteins that inhibit various steps of viral replication as well as facilitate the subsequent activation of acquired immune responses. In this study, we investigated the expression of non-coding RNA upon viral infection or RLR activation. Using a microarray, we identified several microRNAs (miRNA) specifically induced to express by RLR signaling. As suggested by Bioinformatics (miRBase Target Data base), one of the RLR-inducible miRNAs, miR-23b, actually knocked down the expression of very low density lipoprotein receptor (VLDLR) and LDLR-related protein 5 (LRP5). Transfection of miR-23b specifically inhibited infection of rhinovirus 1B (RV1B), which utilizes the low density lipoprotein receptor (LDLR) family for viral entry. Conversely, introduction of anti-miRNA-23b enhanced the viral yield. Knockdown experiments using small interfering RNA (siRNA) revealed that VLDLR, but not LRP5, is critical for an efficient infection by RV1B. Furthermore, experiments with the transfection of infectious viral RNA revealed that miR-23b did not affect post-entry viral replication. Our results strongly suggest that RIG-I signaling results in the inhibitions of infections of RV1B through the miR-23b-mediated down-regulation of its receptor VLDLR.

ApoER2 and VLDLR in the developing human telencephalon

The Reelin-Dab1 signaling pathway plays a crucial role in regulating the migration and position of cortical neurons during the development of the cerebral cortex. Mutation in Reelin may result in severe developmental disorders such as autosomal recessive lissencephaly. Apolipoprotein E receptor type-2 (ApoER2) and very low-density lipoprotein receptor (VLDLR) are canonical receptors of Reelin, through which extracellular Reelin activates the intracellular adapter, Disabled1(Dab1), and subsequently interacts with other molecules. Although it is widely accepted that ApoER2 and VLDLR are indispensable components of the Reelin signaling pathway, little is known of their expression pattern in the laminated developing human brain. Here, we collected 18 cases of human fetal brains of 6–18 gestational weeks (GW) old and examined the expression of ApoER2 and VLDLR in the their telencephalon using immunocytochemical staining. We found that both receptors were absent in the preplate (PP) and the earliest stage of the cortical plate (CP). In later stages of CP development, ApoER2 was expressed earlier than VLDLR in the migrating neurons. Thus, the Reelin-Dab1 signaling pathway may not be involved in the formation of the preplate and deep layers of the CP. Instead, the pathway may act on neurons that are destined to form the more superficial layers of the CP. In addition, the pathway required ApoER2 only rather than both ApoER2 and VLDLR at the initiation of activity.

Abstract LB-219: ApoE and its' receptors (LRP8, VLDLR) function as growth signals for triple-negative breast cancer and represents novel therapeutic targets

Abstract Background: Triple-negative breast cancers (TNBC) lack the expression of estrogen, progesterone, and the human epidermal growth factor 2 receptors and have limited treatment options. We hypothesized that genes that are frequently amplified or overexpressed in these cancers are functionally important for the growth and survival of triple-negative breast cancer. Materials/Methods: We identified genes overexpressed in TNBC in 3 different human breast cancer gene expression data sets (n=294, n=286, n=198). We assessed the functional importance of the consistently overexpressed genes using siRNA screen on 18 breast cancer cell lines (10 ER−, 8 ER+). We targeted each gene with 4 different siRNAs separately and each experiment was performed in triplicate. The genes with the greatest TNBC-specific inhibitory effect after siRNA down regulation were selected for further mechanistic studies. Results: 684 genes showed consistent and highly significant overexpression in TNBC compared to receptor-positive cancers in all 3 data sets (11% of these mapped to DNA segments with frequent copy number gains). 27 genes showed similar effect with 2 or more siRNAs and for 2 genes 3 of the 4 siRNAs showed preferential growth inhibition in ER-negative cells. These two genes were VLDLR (very low-density lipoprotein receptor) and LRP8 (low density lipoprotein receptor-related protein 8). We validated the siRNA screen results and confirmed down regulation of the mRNA and protein levels for VLDLR and LRP8 in 4 different ER-negative cell lines and showed that siRNA inhibition can be rescued by co-transfection of the receptor genes. Reanalysis of gene expression data also indicated expression of the LRP8 and VLDLR ligands, Reelin or ApoE, in both breast cancer tissues and in cell lines. We next demonstrated that exposure to Reelin and ApoE stimulates the growth of ER-negative cells in vitro. The stimulatory effect of ApoE was isoform dependent; ApoE2 (Cys112/Cys158) has the lowest receptor binding affinity and showed no growth stimulation, ApoE3 (Cys112/Arg158) had modest 50–60% growth stimulation and ApoE4 (Arg112/Arg158) had the greatest stimulatory effect (300–400%). Suppression of the expression of either LRP8 or VLDLR or exposure to RAP (an inhibitor of ligand binding to LRP8 and VLDLR) abolished this ligand-induced proliferation. Conclusions: We show that the ApoE -LRP8/VLDLR ligand receptor system is overexpressed in human TNBC. We also demonstrated that this receptor system mediates a strong growth promoting and survival function in TNBC cells in vitro. Interestingly, allelic imbalance favoring ApoE4 expression (E3/E4 or E4/E4) has been linked to higher risk of developing early onset breast cancer, that is primarily TNBC. We propose that inhibitors of LRP8/VLDLR signaling may be clinically useful therapeutic agents for TNBC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-219. doi:10.1158/1538-7445.AM2011-LB-219

Molecular cloning of the two very low-density lipoprotein receptor (VLDLR) subtypes in geese and the effect of overfeeding on their MRNA levels

The objectives of this study were to verify the existence of two subtypes of the very low-density lipoprotein receptor (VLDLR) gene in geese, to investigate the effect of overfeeding on the plasma concentration of triglycerides (TG) and the very low density lipoprotein (VLDL), the activity of lipoprotein lipase (LPL), and the mRNA level of VLDLR in Sichuan White geese and Landes geese. The results indicate that there are two subtypes of the VLDLR gene in geese, and that they share a high similarity with those of other species. The expression of VLDLR I and VLDLR II was found in both tissues examined. After overfeeding, the expression level of VLDLR I in adipose tissue showed about a onefold increase (P &lt; 0.05) in both breeds. Overfeeding induced a significant decrease of VLDLR I in skeletal muscle of both breeds, and a significant decrease of VLDLR II in Sichuan White geese (P &lt; 0.05), but an obvious increase of VLDLR II in Landes geese (P &lt; 0.05). In addition, overfeeding induced the increase of plasma VLDL, TG concentration and plasma LPL activity. It was concluded that VLDLR may participate in the metabolism of VLDL-TG by regulating the LPL-mediated TG hydrolysis in geese. Key words: Gene expression, geese, molecular cloning, overfeeding, very low-density lipoprotein receptor

Peroxisome Proliferator-activated Receptor γ Agonist Rosiglitazone Increases Expression of Very Low Density Lipoprotein Receptor Gene in Adipocytes

Apolipoprotein E (apoE) and its receptor, very low density lipoprotein receptor (VLDLR), are involved in fat accumulation in adipocytes. Here, we investigated the effect of a peroxisome proliferator-activated receptor (PPAR) gamma agonist, rosiglitazone, on regulation of VLDLR expression both in white adipose tissue (WAT) of obese mice and in cultured adipocytes. Furthermore, to determine whether rosiglitazone directly regulates transcription of the VLDLR gene, we carried out luciferase assay with a reporter gene containing mouse VLDLR promoter region, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay. Four-day treatment with rosiglitazone increased the expression of VLDLR in WAT of ob/ob mice. Moreover, rosiglitazone increased the expression of VLDLR in cultured adipocytes. The PPAR-responsive element (PPRE)-directed mutagenesis analyses revealed that the PPRE motif in the VLDLR promoter region plays a significant role in transcriptional activation of the VLDLR gene in adipocytes. In addition, electrophoretic mobility shift assay and chromatin immunoprecipitation assay demonstrated that endogenous PPARgamma directly binds to this functional PPRE motif in the VLDLR promoter region. We also investigated the effects of rosiglitazone on insulin sensitivity and lipid accumulation in both ob/ob mice and apoE-deficient ob/ob mice. Rosiglitazone ameliorated insulin sensitivity in both ob/ob mice and apoE-deficient ob/ob mice, possibly through decreasing the expression of monocyte chemoattractant protein-1 (MCP-1), increasing the expression of superoxide dismutase 1 (SOD1) in WAT, and increasing plasma adiponectin concentration. In ob/ob mice, body weight and WAT weight were significantly higher in the mice treated with rosiglitazone than those treated with vehicle. However, in apoE-deficient ob/ob mice, no significant difference in body weight or WAT weight was observed between the vehicle-treated group and the rosiglitazone-treated group. Moreover, rosiglitazone did not increase body weight and WAT weight in VLDLR-deficient mice. These findings indicate that rosiglitazone directly increases VLDLR expression, thereby enhancing apoE-VLDLR-dependent lipid accumulation in adipocytes.

Overproduced interleukin 6 decreases blood lipid levels via upregulation of very-low-density lipoprotein receptor

BackgroundInterleukin 6 (IL6) blockade raises blood lipid levels in patients with rheumatoid arthritis.ObjectiveTo examine the influence of IL6 on lipid metabolism.MethodsVascular smooth muscle cells (VSMC) were cultured in the presence...

Expression of Scavenger Receptor-BI and Low-Density Lipoprotein Receptor and Differential Use of Lipoproteins to Support Early Steroidogenesis in Luteinizing Macaque Granulosa Cells

An ovulatory hCG stimulus to rhesus macaques undergoing controlled ovarian stimulation protocols results in a rapid and sustained increase in progesterone synthesis. The use of lipoproteins as a substrate for progesterone synthesis remains unclear, and the expression of lipoprotein receptors [very-low-density lipoprotein receptor (VLDLR), low-density lipoprotein receptor (LDLR), and scavenger receptor-BI (SR-BI)] soon after human chorionic gonadotropin (hCG) (<12 h) has not been characterized. This study investigated lipoprotein receptor expression and lipoprotein (VLDL, LDL, and HDL) support of steroidogenesis during luteinization of macaque granulosa cells. Granulosa cells were aspirated from rhesus monkeys undergoing controlled ovarian stimulation before or up to 24 h after an ovulatory hCG stimulus. The expression of VLDLR decreased within 3 h of hCG, whereas LDLR and SR-BI increased at 3 and 12 h, respectively. Granulosa cells isolated before hCG were cultured for 24 h in the presence of FSH or FSH plus hCG with or without VLDL, LDL, or HDL. Progesterone levels increased in the presence of hCG regardless of lipoprotein addition, although LDL, but not HDL, further augmented hCG-induced progesterone. Other cells were cultured with FSH or FSH plus hCG without an exogenous source of lipoprotein for 24 h, followed by an additional 24 h culture with or without lipoproteins. Cells treated with hCG in the absence of any lipoprotein were unable to maintain progesterone levels through 48 h, whereas LDL (but not HDL) sustained progesterone synthesis. These data suggest that an ovulatory stimulus rapidly mobilizes stored cholesterol esters for use as a progesterone substrate and that as these are depleted, new cholesterol esters are obtained through an LDLR- and/or SR-BI-mediated mechanism.

Glucose deprivation accelerates VLDL receptor-mediated TG-rich lipoprotein uptake by AMPK activation in skeletal muscle cells

Glucose and fatty acids are major energy sources in skeletal muscle. Very low-density lipoprotein receptor (VLDL-R), which is highly expressed in heart, skeletal muscle and adipose tissue, plays a crucial role in metabolism of triglyceride (TG)-rich lipoproteins. To explore energy switching between glucose and fatty acids, we studied expression of VLDL-R and lipoprotein uptake in rat L6 myoblasts. l-Glucose or d-glucose deprivation in the medium noticeably induced the AMPK (AMP-activated protein kinase) activation and VLDL-R expression. Dose-dependent induction of VLDL-R expression was observed when d-glucose was less than 4.2mM. The same phenomenon was also observed in rat primary skeletal myoblasts and cultured vascular smooth muscle cells. The uptake of β-VLDL but not LDL was accompanied by induction of VLDL-R expression. Our study suggests that the VLDL-R-mediated uptake of TG-rich lipoproteins might compensate for glucose shortfall through AMPK activation in skeletal muscle.