SR-BI Expression Research Articles

Phaleria macrocarpa Boerl. (Thymelaeaceae) leaves increase SR-BI expression and reduce cholesterol levels in rats fed a high cholesterol diet.

In vitro and in vivo studies of the activity of Phaleria macrocarpa Boerl (Thymelaeaceae) leaves against the therapeutic target for hypercholesterolemia were done using the HDL receptor (SR-BI) and hypercholesterolemia-induced Sprague Dawley rats. The in vitro study showed that the active fraction (CF6) obtained from the ethyl acetate extract (EMD) and its component 2',6',4-trihydroxy-4'-methoxybenzophenone increased the SR-BI expression by 95% and 60%, respectively. The in vivo study has proven the effect of EMD at 0.5 g/kgbw dosage in reducing the total cholesterol level by 224.9% and increasing the HDL cholesterol level by 157% compared to the cholesterol group. In the toxicity study, serum glutamate oxalate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) activity were observed to be at normal levels. The liver histology also proved no toxicity and abnormalities in any of the treatment groups, so it can be categorized as non-toxic to the rat liver. The findings taken together show that P. macrocarpa leaves are safe and suitable as an alternative control and prevention treatment for hypercholesterolemia in Sprague Dawley rats.

A Western-fed diet increases plasma HDL and LDL-cholesterol levels in apoD-/- mice.

ObjectivePlasma apolipoprotein (apo)D, a ubiquitously expressed protein that binds small hydrophobic ligands, is found mainly on HDL particles. According to studies of human genetics and lipid disorders, plasma apoD levels positively correlate with HDL-cholesterol and apoAI levels. Thus, we tested the hypothesis that apoD was a regulator of HDL metabolism.Methods & ResultsWe compared the plasma lipid and lipoprotein profiles of wild-type (WT) C57BL/6 mice with apoD−/− mice on a C57BL/6 background after receiving a high fat-high cholesterol diet for 12 weeks. ApoD−/− mice had higher HDL-cholesterol levels (61±13-apoD−/− vs. 52±10-WT-males; 37±11-apoD−/− vs. 22±2 WT-female) than WT mice with sex-specific changes in total plasma levels of cholesterol and other lipids. Compared to WT, the HDL of apoD−/− mice showed an increase in large, lipid-rich HDL particles and according to size various quantities and sizes of LDL particles. Plasma levels of lecithin:cholesterol acyltransferase in the control and apoD−/− mice were not different, however, plasma phospholipid transfer protein activity was modestly elevated (+10%) only in male apoD−/− mice. An in vivo HDL metabolism experiment with isolated Western-fed apoD−/− HDL particles showed that female apoD−/− mice had a 36% decrease in the fractional catabolic rate of HDL cholesteryl ester. Hepatic SR-BI and LDLR protein levels were significantly decreased; accordingly, LDL-cholesterol and apoB levels were increased in female mice.ConclusionIn the context of a high fat-high cholesterol diet, apoD deficiency in female mice is associated with increases in both plasma HDL and LDL-cholesterol levels, reflecting changes in expression of SR-BI and LDL receptors, which may impact diet-induced atherosclerosis.

Mipu1 overexpression protects macrophages from oxLDL-induced foam cell formation and cell apoptosis.

Mipu1 (myocardial ischemic preconditioning upregulated protein 1) is a novel N-terminal Kruppel-associated box (KRAB)/C2H2 zinc finger superfamily protein, that displays a powerful effect in protecting H9c2 cells from oxidative stress-induced cell apoptosis. The present study aims to investigate the effect of Mipu1 overexpression on oxidized low-density lipoprotein (oxLDL)-induced foam cell formation, cell apoptosis, and its possible mechanisms. New Zealand healthy rabbits were used to establish atherosclerosis model, and serum levels of triglycerides, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were detected by an automatic biochemical analyzer. Sudan IV staining was used to detect atherosclerotic lesions. The RAW264.7 macrophage cell line was selected as the experimental material. Oil red O staining, high-performance liquid chromatography, and Dil-labeled lipoprotein were used to detect cholesterol accumulation qualitatively and quantitatively, respectively. Flow cytometry was used to determine cell apoptosis. Real-time quantitative polymerase chain reaction (PCR) was used to detect the mRNA expression of the main proteins that are associated with the transport of cholesterol, such as ABCA1, ABCG1, SR-BI, and CD36. Western blot analysis was used to detect the protein expression of Mipu1. There were atherosclerotic lesions in the high-fat diet group with Sudan IV staining. High-fat diet decreased Mipu1 expression and increased CD36 expression significantly at the 10th week compared with standard-diet rabbits. Mipu1 overexpression decreased oxLDL-induced cholesterol accumulation, oxLDL uptake, cell apoptosis, and cleaved caspase-3. Mipu1 overexpression inhibited the oxLDL-induced CD36 mRNA and protein expression, but it did not significantly inhibit the mRNA expression of ABCA1, ABCG1, and SR-BI. Mipu1 overexpression inhibits oxLDL-induced foam cell formation and cell apoptosis. Mipu1 overexpression reduces the lipid intake of macrophages and might be associated with the downregulation of CD36 expression in the presence of oxLDL.

Liver growth factor induces testicular regeneration in EDS-treated rats and increases protein levels of class B scavenger receptors.

The aim of the present work was to determine the effects of liver growth factor (LGF) on the regeneration process of rat testes after chemical castration induced by ethane dimethanesulfonate (EDS) by analyzing some of the most relevant proteins involved in cholesterol metabolism, such as hormone sensitive lipase (HSL), 3β-hydroxysteroid dehydrogenase (3β-HSD), scavenger receptor SR-BI, and other components of the SR family that could contribute to the recovery of steroidogenesis and spermatogenesis in the testis. Sixty male rats were randomized to nontreated (controls) and LGF-treated, EDS-treated, and EDS + LGF-treated groups. Testes were obtained on days 10 (T1), 21 (T2), and 35 (T3) after EDS treatment, embedded in paraffin, and analyzed by immunohistochemistry and Western blot. LGF improved the recovery of the seminiferous epithelia, the appearance of the mature pattern of Leydig cell interstitial distribution, and the expression of mature SR-BI. Moreover, LGF treatment resulted in partial recovery of HSL expression in Leydig cells and spermatogonia. No changes in serum testosterone were observed in control or LGF-treated rats, but in EDS-castrated animals LGF treatment induced a progressive increase in serum testosterone levels and 3β-HSD expression. Based on the pivotal role of SR-BI in the uptake of cholesteryl esters from HDL, it is suggested that the observed effects of LGF would facilitate the provision of cholesterol for sperm cell growth and Leydig cell recovery.

GW25-e0743 Effects of Astragalus Polysaccharides on the expression of SR-BI, LXRα in RAW264.7 macrophage-derived foam cells

GW25-e0743 Effects of Astragalus Polysaccharides on the expression of SR-BI, LXRα in RAW264.7 macrophage-derived foam cells

Original article. Patterns of microglial innate immune responses elicited by amyloid β1–42and lipopolysaccharide: the similarities of the differences

AbstractBackground: As part of their innate immune response to changes in the central nervous system environment, normally quiescent microglia become activated and increase expression of pattern recognition receptors, scavenger receptors, and production of inflammatory cytokines, proteinases, reactive oxygen species (ROS), and free radicals. These molecules have been implicated in the pathogenesis and progression of several neurodegenerative disorders including Alzheimer disease (AD).Objective: We compared patterns of microglial innate immune responses elicited by nonfibrillar amyloid β peptide (nfAβ1-42) to those elicited by lipopolysaccharide (LPS).Methods: Murine BV-2 microglial cells were exposed to either nfAβ1-42or LPS for 12 h. Then, total RNA from each condition was isolated and expression levels of Toll-like receptor (TLR)-4, scavenger receptor class A (SRMARCO) and class B (SR-BI), CD36, and matrix metalloproteinase (MMP)-9 were determined by reverse transcription-quantitative real-time polymerase chain reaction. The amount of hydrogen peroxide (H2O2) and nitric oxide (NO) in the cell-free supernatant at 24 h were determined using 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red) and Griess reagent, respectively.Results: nfAβ1-42and LPS significantly increased expression of TLR-4, SR-MARCO, CD36, and MMP-9 and production of H2O2 and NO in BV-2 microglial cells compared with that of unstimulated cells. However, expression of SR-BI was significantly induced only when the cells were exposed to nfAβ1-42.Conclusion: These findings indicate that pattern of microglial innate immune responses elicited by nfAβ1-42overlap with that elicited by LPS and suggest a specific role of microglial SR-BI expression in AD pathogenesis.

Chronic unpredictable mild stress combined with a high-fat diets aggravates atherosclerosis in rats.

BackgroundDepression and high-fat diet are both known as independent risk factors for atherosclerosis and other cardiovascular diseases, suggesting the interaction of psychological and physiological factors in the development of these diseases. The liver is a crucial organ that facilitate lipid metabolism especially in reverse cholesterol transport (RCT), while according to the theory of Traditional Chinese Medicine, depression as a kind of psychological stress has an influence on hepatic function. So there seem to be some links between depression and lipid metabolic disorders.MethodsTo investigate these links, we separately treated rats with chronic unpredictable mild stress (CMS) and/or a high-fat diet (HD) to evaluate the development of atherosclerosis and the expression of hepatic ABCG8, ABCG5, SR-BI, CYP7A1, LXRα, and LCAT which were associated with reverse cholesterol transport.ResultsThis study provided evidence that high-fat diet greatly decreased these genes expression related to RCT while chronic stress alone tended to promote RCT. Chronic unpredictable mild stress combined with a high-fat diet attenuated RCT and aggravated atherogenesis.ConclusionsThese observations suggested that chronic psychological stress alone is virtually propitious to lipid metabolism, however when under a condition of high-fat diet, it deteriorated atherosclerotic plague and did harm to RCT.

Abstract 32: Liver-Specific Transgenic Expression of Cholesteryl Ester Hydrolase Reduces Atherosclerosis in LDLR-/- Mice

Liver plays a central role in the final elimination of cholesterol from the body either as bile acids or as free cholesterol (FC) and lipoprotein cholesterol is the major source of total biliary cholesterol. HDL is the major lipoprotein responsible for removal and transport of cholesterol mainly as cholesteryl esters (CE) from the peripheral tissues to the liver. While HDL-FC is rapidly secreted into bile, the fate of HDL-CE remains unclear. We have earlier demonstrated the role of human cholesteryl ester hydrolase (CEH, CES1) in hepatic hydrolysis of HDL-CE and increasing bile acid synthesis, a process dependent on SR-BI expression. In the present study, we examined the hypothesis that by enhancing the elimination of HDL-CE into bile/feces liver-specific transgenic expression of CEH will be anti-atherogenic. Increased CEH expression in the liver led to significant increase in flux of HDL-CE to bile acids. In LDLR-/- background, this enhanced elimination of cholesterol led to attenuation of diet-induced atherosclerosis in LDLR-/-LCEH2 mice (Figure) with a consistent increase in fecal sterol secretion primarily as bile acids. Taken together with the observed reduction in atherosclerosis by increasing macrophage CEH-mediated cholesterol efflux, these studies establish CEH as an important regulator in enhancing cholesterol elimination and also as an anti-atherogenic target.

Abstract 669: SR-BI in Bone Marrow--Derived Cells Protects Mice From Diet-Induced Coronary Artery Atherosclerosis and Myocardial Infarction

Background: SR-BI deficient, apolipoprotein E hypomorphic mice develop diet induced occlusive coronary artery atherosclerosis, myocardial infarction and early death. Methods: To test the role of SR-BI in bone marrow derived cells, SR-BI-null; apoE- hypomorphic mice were transplanted with SR-BI +/+ apoE-hypomorphic, or control, autologous SR-BI-null; apoE-hypomorphic bone marrow. Four weeks later, mice were fed a high-fat, high-cholesterol, cholate-containing diet to induce coronary artery atherosclerosis. Results: Mice transplanted with autologous bone marrow developed extensive aortic atherosclerosis and severe occlusive coronary artery atherosclerosis after 4 weeks of feeding. This was accompanied by myocardial fibrosis and increased heart weights. In contrast, restoration of SR-BI expression in bone marrow derived-cells reduced diet induced aortic and coronary artery atherosclerosis, myocardial fibrosis and the increase in heart weights in SR-BI-null; apoE-hypomorphic mice. Restoration of SR-BI in bone marrow derived cells did not, however, affect steady state lipoprotein cholesterol levels, but reduced plasma levels of IL-6. Monocytes from SR-BI-null mice exhibited a greater capacity to bind to VCAM-1 and ICAM-1 than those from SR-BI +/+ mice. Furthermore, restoration of SR-BI expression in bone marrow derived cells attenuated monocyte recruitment into atherosclerotic plaques in mice fed high fat, high cholesterol cholate containing diet. Conclusion: These data demonstrate directly that SR-BI in bone marrow-derived cells protects against both aortic and coronary artery atherosclerosis.

Inhibition of mTOR down-regulates scavenger receptor, class B, type I (SR-BI) expression, reduces endothelial cell migration and impairs nitric oxide production

The mammalian target of rapamycin (mTOR) inhibiting drug rapamycin (Sirolimus) has severe side effects in patients including hyperlipidemia, an established risk factor for atherosclerosis. Recently, it was shown that rapamycin decreases hepatic LDL receptor (LDL-R) expression, which likely contributes to hypercholesterolemia. Scavenger receptor, class B, type I (SR-BI) is the major HDL receptor and consequently regulating HDL-cholesterol levels and the athero-protective effects of HDL. By using the mTOR inhibitor rapamycin, we show that SR-BI is down-regulated in human umbilical vein endothelial cells (HUVECs). This reduction of SR-BI protein as well as mRNA levels by about 50% did not alter HDL particle uptake or HDL-derived lipid transfer. However, rapamycin reduced HDL-induced activation of eNOS and stimulation of endothelial cell migration. The effects on cell migration could be counteracted by SR-BI overexpression, indicating that decreased SR-BI expression is in part responsible for the rapamycin-induced effects. We demonstrate that inhibition of mTOR leads to endothelial cell dysfunction and decreased SR-BI expression, which may contribute to atherogenesis during rapamycin treatment.

GABA and Topiramate Inhibit the Formation of Human Macrophage-Derived Foam Cells by Modulating Cholesterol-Metabolism-Associated Molecules

Aims: γ-aminobutyric acid (GABA), the principal inhibitory neurotransmitter, acts on GABA receptors to play an important role in the modulation of macrophage functions. The present study examined the effects of GABA and a GABA receptor agonist on modulating cholesterol-metabolism-associated molecules in human monocyte-derived macrophages (HMDMs). Methods: ORO stain, HPLC, qRT-PCR, Western blot and EMSA were carried out using HMDMs exposed to ox-LDL with or without GABAergic agents as the experimental model. Results: GABA and topiramate reduced the percentage of cholesterol ester in lipid-laden HMDMs by down-regulating SR-A, CD36 and LOX-1 expression and up-regulating ABCA1, ABCG1 and SR-BI expression in lipid-laden HMDMs. The production of TNF-a was decreased in GABA-and topiramate-treated lipid-laden HMDMs, and levels of interleukin (IL)-6 did not change. The activation of two signaling pathways, p38MAPK and NF-γB, was repressed by GABA and topiramate in lipid-laden HMDMs. Conclusion: GABA and topiramate inhibit the formation of human macrophage-derived foam cells and may be a possibility for macrophage targeted therapy of atherosclerotic lesions.

Abstract 18570: Intronic SCARB1 (Scavenger Receptor Class B Type I, SR-BI) rs10846744 Variant Mediates Functional Effect by Enhancer Activity

Objective: We previously reported the significant association of SCARB1 intronic rs10846744 in reducing odds ratio for incident coronary heart disease (CHD) in participants of the Multi-Ethnic Study of Atherosclerosis. Using in vitro and ex vivo approaches, we sought to define the underlining molecular mechanism for this clinical association. Methods and results: Using publicly available genome databases, a bioinformatic screen revealed that rs10846744 resided in a region exhibiting DNase I hypersensitivity and H3K4me1 chromatin methylation markers. Gel shift assays revealed significantly lower binding of nuclear proteins to the probe expressing the risk C allele for CHD ( p <0.001). Yeast-One Hybrid screening revealed NR2F2 as a candidate transcription factor (TF), while bioinformatic screens (HaploReg v2) predicted binding of multiple TFs such as AP2[[Unable to Display Character: 𝛄]], Max, ERα, FOXA1, GATA3 and p300 to this region. Using RNA Seq, we confirmed that lymphocytes isolated from hyperalphalipoproteinemic (HALP) carriers of the risk C allele had significantly higher NR2F2 expression (6.3-fold upregulated, p <0.001). Interestingly, under basal conditions rs10846744 did not significantly regulate SR-BI RNA and protein expression in these lymphocytes. Global metabolic profiling from plasma samples of HALP carriers of the risk C allele had significantly higher markers of oxidative stress (42% higher cysteinyl-glycine, oxidized, p =0.0136, q =0.2437) and lower levels of lysolipids (46% lower, p =0.0043, q =0.1605). Conclusion: The intronic rs10846744 variant resides within a functionally active regulatory region of SCARB1 , without directly regulating SR-BI expression itself. Our results confirm the importance of dissecting the pathway by which this region influences CHD.

SR-BI in Bone Marrow Derived Cells Protects Mice from Diet Induced Coronary Artery Atherosclerosis and Myocardial Infarction

SR-BI deficient mice that are also hypomorphic for apolipoprotein E expression develop diet induced occlusive coronary artery atherosclerosis, myocardial infarction and early death. To test the role of SR-BI in bone marrow derived cells, we used bone marrow transplantation to generate SR-BI-null; apoE-hypomorphic mice in which SR-BI expression was restored solely in bone marrow derived cells. SR-BI-null; apoE-hypomorphic mice were transplanted with SR-BI+/+apoE-hypomorphic, or control, autologous SR-BI-null; apoE-hypomorphic bone marrow. Four weeks later, mice were fed a high-fat, high-cholesterol, cholate-containing diet to induce coronary artery atherosclerosis. Mice transplanted with autologous bone marrow developed extensive aortic atherosclerosis and severe occlusive coronary artery atherosclerosis after 4 weeks of feeding. This was accompanied by myocardial fibrosis and increased heart weights. In contrast, restoration of SR-BI expression in bone marrow derived-cells reduced diet induced aortic and coronary artery atherosclerosis, myocardial fibrosis and the increase in heart weights in SR-BI-null; apoE-hypomorphic mice. Restoration of SR-BI in bone marrow derived cells did not, however, affect steady state lipoprotein cholesterol levels, but did reduce plasma levels of IL-6. Monocytes from SR-BI-null mice exhibited a greater capacity to bind to VCAM-1 and ICAM-1 than those from SR-BI+/+ mice. Furthermore, restoration of SR-BI expression in bone marrow derived cells attenuated monocyte recruitment into atherosclerotic plaques in mice fed high fat, high cholesterol cholate containing diet. These data demonstrate directly that SR-BI in bone marrow-derived cells protects against both aortic and CA atherosclerosis.

GW24-e0042 Effects of AngII and Ang-(1-7) on the expression of SR-BI, ABCA1 and cholesterol efflux in THP-1 derived foam cells

ObjectivesTo investigate the effects of angiotensinII (AngII) and angiotensin-(1-7) [Ang-(1-7)] on scavenger receptor class B type1 (SR-BI), ATP-binding cassette transporter A1 (ABCA1) and cholesterol efflux in THP-1 derived foam cells.MethodsHuman...

GW24-e0058 Effects of SR-BI Deficiency and Hypercholesterolemia on Erythropoiesis

ObjectivesScavenger receptor BI (SR-BI) is a key regulator of high density lipoprotein (HDL) metabolism. Recent studies showed that mice deficient in SR-BI exhibit impaired erythropoiesis and it has been proposed...

Challenges in Using Cultured Primary Rodent Hepatocytes or Cell Lines to Study Hepatic HDL Receptor SR-BI Regulation by Its Cytoplasmic Adaptor PDZK1

BackgroundPDZK1 is a four PDZ-domain containing cytoplasmic protein that binds to a variety of membrane proteins via their C-termini and can influence the abundance, localization and/or function of its target proteins. One of these targets in hepatocytes in vivo is the HDL receptor SR-BI. Normal hepatic expression of SR-BI protein requires PDZK1 - <5% of normal hepatic SR-BI is seen in the livers of PDZK1 knockout mice. Progress has been made in identifying features of PDZK1 required to control hepatic SR-BI in vivo using hepatic expression of wild-type and mutant forms of PDZK1 in wild-type and PDZK1 KO transgenic mice. Such in vivo studies are time consuming and expensive, and cannot readily be used to explore many features of the underlying molecular and cellular mechanisms.Methodology/Principal FindingsHere we have explored the potential to use either primary rodent hepatocytes in culture using 2D collagen gels with newly developed optimized conditions or PDZK1/SR-BI co-transfected cultured cell lines (COS, HEK293) for such studies. SR-BI and PDZK1 protein and mRNA expression levels fell rapidly in primary hepatocyte cultures, indicating this system does not adequately mimic hepatocytes in vivo for analysis of the PDZK1 dependence of SR-BI. Although PDZK1 did alter SR-BI protein expression in the cell lines, its influence was independent of SR-BI’s C-terminus, and thus is not likely to occur via the same mechanism as that which occurs in hepatocytes in vivo.Conclusions/SignificanceCaution must be exercised in using primary hepatocytes or cultured cell lines when studying the mechanism underlying the regulation of hepatic SR-BI by PDZK1. It may be possible to use SR-BI and PDZK1 expression as sensitive markers for the in vivo-like state of hepatocytes to further improve primary hepatocyte cell culture conditions.

Porphyromonas gingivalis lipopolysaccharide increases lipid accumulation by affecting CD36 and ATP-binding cassette transporter A1 in macrophages

Porphyromonas gingivalis lipopolysaccharide (P. gingivalis LPS) promotes macrophage-derived foam cell formation, however, the mechanisms are not well established. In macrophages, lipid uptake is mediated by scavenger receptors including SR-A and CD36, while the cholesterol efflux is mediated by ATP-binding cassette transporter G1 (ABCG1), ABCA1 and SR-BI. We further investigated the mechanisms underlying the dysregulation by P. gingivalis LPS of these regulators resulting in the promotion of lipid accumulation in THP-1-derived macrophages. Our results showed that P. gingivalis LPS exacerbated lipid accumulation in oxidized low-density lipoprotein (oxLDL)-treated macrophages. However, cholesterol efflux was inhibited by P. gingivalis LPS in THP-1-derived macrophages. In oxLDL-untreated macrophages, P. gingivalis LPS treatment caused an increase in CD36 mRNA and protein levels, and a decrease in ABCA1 mRNA and protein levels, while having no effect on SR-A, SR-BI or ABCG1 expression. Upregulation of CD36 by P. gingivalis LPS resulted from activation of c-Jun/AP-1, and this was confirmed by the inhibition of increased CD36 expression after AP-1 inhibition using SP600125. However, the decreased protein stability of ABCA1 by P. gingivalis LPS was a result of increased calpain activity. Moreover, small hairpin RNA (shRNA) targeting heme oxygenase-1 (HO-1) augmented the P. gingivalis LPS-induced atherogenic effects on the expression of c-Jun/AP-1, CD36, ABCA1 and calpain activity. Accordingly, P. gingivalis LPS-regulated promotion of lipid accumulation in foam cells was also exacerbated by HO-1 shRNA. These results indicate that P. gingivalis LPS confers a exacerbation effect on the formation of foam cells by a novel HO-1-dependent mediation of cholesterol efflux and lipid accumulation in macrophages.

Excess nitric oxide impairs liver X receptor α-ATP-binding cassette transporter A1-dependent cholesterol efflux in macrophage foam cells

Excess nitric oxide (NO) promotes the progression of atherosclerosis by increasing the oxidation of low-density lipoprotein (LDL) and inflammatory responses. However, little is known about the impact of NO and its underlying molecular mechanism on lipid metabolism of macrophage foam cells. In this study, Oil-red O staining, cholesterol and triglyceride assay, Dil-oxidized LDL (oxLDL) binding assay, cholesterol efflux assay, real-time RT-PCR and Western blot analysis were used for in vitro experiments. Apolipoprotein E-deficient (apoE(-/-) ) and apoE and inducible nitric oxide synthase-deficient (apoE(-/-) iNOS(-/-) ) mice were as our in vivo models. Treatment with S-nitroso-N-acetyl-D,L-penicillamine (SNAP), an NO donor, exacerbated oxLDL-induced cholesterol accumulation in macrophages, because of reduced efficacy of cholesterol efflux. In addition, SNAP decreased the protein level of ATP-binding cassette transporter A1 (ABCA1) without affecting scavenger receptor type A (SR-A), CD36, ABCG1, or SR-B1 levels. This SNAP-mediated downregulation of ABCA1 was mainly through the effect of NO but not peroxynitrite. Furthermore, the SNAP-downregulated ABCA1 was due to the decrease in the liver X receptor α (LXRα)-dependent transcriptional regulation. Moreover, genetic deletion of iNOS increased the serum capacity of reverse cholesterol efflux and protein expression of LXRα, ABCA1, and SR-BI in aortas and retarded atherosclerosis in apoE(-/-) mice. Our findings provide new insights in the pro-atherogenic effect of excess NO on cholesterol metabolism in macrophages.

Nifedipine Enhances Cholesterol Efflux in RAW264.7 Macrophages

Studies have shown that nifedipine protects against atherosclerotic progression, but its underlying mechanisms remain unclear. In this study, we examined if nifedipine increases macrophage cholesterol efflux, a pathway known to inhibit atherogenesis. We evaluated the ability of different doses of nifedipine to affect cholesterol efflux in RAW264.7 macrophages and its relationship with mRNA and protein levels of several well-characterized proteins involved in cholesterol efflux, including ABCA1, ABCG1, SR-BI and LXRα, using quantitative real-time PCR, Western blotting, and siRNA techniques. Nifedipne at 1, 10, and 100 nmol/L increased apoA-I-mediated cholesterol efflux from 2.55 % to 5.65 %, 6.20 %, and 6.10 %, as well as HDL-mediated cholesterol efflux from 31.0 % to 42.5 %, 46.0 %, and 43.5 %, respectively, in RAW264.7 macrophages (p < 0.05), which was associated with increased mRNA expression levels of ABCA1, ABCG1, SR-BI, and LXRα (405 %, 381 %, 336 %; 890 %, 960 %, 1002 %; 285 %, 325 %, 336 %; 482 %, 445 %, 405 %, respectively, p < 0.05), and with increased protein levels of ABCA1, ABCG1, SR-BI, and LXRα (428 %, 492 %, 361 %; 288 %, 331 %, 365 %; 283 %, 320 %, 505 %; 581 %, 678 %, 608 %, respectively, p < 0.05). SiRNA-mediated silencing of LXRα revealed that LXRα was involved in these increases and the enhanced cholesterol efflux. Nifedipine may protect against atherosclerosis partly by promoting macrophage cholesterol efflux through the stimulation of LXRα-dependent expression of ABCA1, ABCG1, and SR-BI.

ASSA13-03-17 b3-Adrenoceptor Activation Improves the Formation of Atherosclerosis Plaque in Aged ApoE-Deficient Mice

Objective To study the effects of β 3 -adrenoceptor (β 3 -AR) activation on atherosclerotic plaque development in aged apolipoprotein E-deficient (ApoE -/- ) mice. Methods Male ApoE -/- mice were fed with a high-fat diet, starting on 10 weeks of age. Starting from 36 weeks of age, mice were randomly treated with atorvastatin (10 mg · kg -1 ·d -1 , by gavage), the β 3 -AR agonist BRL37344 (1.65 or 3.30 μg·kg -1 , twice a week, by intraperitoneal injection), the β3-AR antagonist SR52390A (50 μg·kg -1 , twice a week, by intraperitoneal injection), or saline vehicle (twice a week, by intraperitoneal injection) for 12 weeks (n = 10 per group). A group of wild-type C57BL/6J mice receiving normal diet was included as a healthy control. At the end of the 12-week treatment period, serum concentration of lipids were measured. Atherosclerotic plaque in the thoracic aorta was examined using Hematoxy-eosin staining. The extent of fibrosis in the plaque was examined using Masson staining. Western blot was uesed to determine the ApoA-I and SR-BI protein expression. Real-time quantitative PCR was used to determine β 3 -AR mRNA in the white adipose tissue and the ApoA-I, ApoA-II and SR-BI mRNA expression in liver tissue. Results High-fat diet in ApoE -/- mice led to hyperlipidemia and the area of and the extent of fibrosis in atherosclerotic plaque (P 3 -AR mRNA and SR-BI mRNA were lower, ApoA-I and ApoA-II mRNA were higher in ApoE -/- mice (P -/- mice receiving vehicle). Treatment with the β 3 -AR agonist BRL37344 decreased serum concentration of triglyceride, total cholesterol and non-high density lipoprotein cholesterol. The treatment increased high density lipoprotein cholesterol, downregulated of ApoA-I, ApoA-II and SR-BI expression and upregulated the β 3 -AR mRNA expression (P -/- mice receiving vehicle). Similar to atorvastatin, BRL37344 treatment dose-dependently reduced the plaque area and collagen content (P -/- mice receiving vehicle at a dose of 3.30 but not 1.65 μg·kg -1 ). Treatment with the β 3 -AR antagonist SR52390A did not affect any parameters (P > 0.05 for all measures vs. ApoE -/- mice receiving vehicle). Conclusions β 3 -AR agonist impeded the progression of atherosclerosis in ApoE -/- mice, possibly through improvement of lipid profile, ApoA-I, ApoA-II and SR-BI mRNA.