共查询到20条相似文献，搜索用时 17 毫秒
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AIM:To study the characteristic of liver X receptor alpha (LXRα), it’s target gene expression and cholesterol efflux from human macrophages in coronary atherosclerotic disease (CAD) patients. METHODS:Human monocyte-derived macrophages from CAD patients and controls were collected. Before being detected apoA-I-mediated human monocyte-derived macrophage cholesterol efflux and LXRα and mRNA expression of its target gene, the macrophages were induced with or without TO-901317. 〖JP2〗RESULTS:Compared with control normal macrophage, the mRNA levels of LXRα and its target gene expression were changed, and the macrophage cholesterol efflux was decreased in CAD patients. After stimulationwith TO-901317, the reactive capacity of LXR was also decreased from human monocyte-derived macrophage of CAD patients. CONCLUSION:The changes of cholesterol efflux and some gene expression in macrophages may be the pathogenetic cause atherosclerosis, and macrophage LXR activity may offer potential therapeutic benefit in the treatment of CAD. 相似文献
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AIM To explore the effects of nicotinic acid (NA) on lysosomal free cholesterol efflux in macrophages and its underlying mechanism. METHODS Macrophages induced from human monocytic leukemia cell line THP-1 by phorbol myristate acetate served as the cell model. Laser scanning confocal microscopy was applied to observe the effects of NA on lysosomal free cholesterol efflux in macrophages loaded with oxidized low-density lipoprotein (oxLDL). The influences of nicotinic acid adenine dinucleotide phosphate (NAADP) antagonist Ned-19, Ca2+ chelator BAPTA, liver X receptor α (LXRα ) siRNA and Niemann-Pick C1 protein (NPC1 ) siRNA on NA effects were also evaluated. RT-qPCR and Western blot were conducted to evaluate the influence of NA, Ned-19 and BAPTA on LXRα mRNA and NPC1 protein expression. RESULTS NA dose-dependently promoted lysosomal free cholesterol efflux in macrophages. This effect was markedly inhibited by Ned-19 and BAPTA. NA increased NPC1 protein and LXRα mRNA expression. These effects were also attenuated by Ned-19 and BAPTA remarkably. LXRα siRNA significantly inhibited the promoting effect of NA on NPC1 protein expression. Silencing of LXRα and NPC1 with siRNA remarkably abolished the effect of NA on lysosomal free cholesterol efflux. CONCLUSION NA promotes lysosomal free cholesterol efflux in macrophages. This effect may be mediated by the increased production of NAADP, which subsequently promotes Ca2+ release through lysosomal transient receptor potential mucolipin 1 (TRPML1) channel and finally up-regulates NPC1 protein expression via LXRα. 相似文献
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AIM: To explore the expression and activation of matrix metalloproteinase-9 (MMP-9) from monocyte-derived macrophages induced by tumor necrosis factor-α (TNF-α) and to investigate its association with progression of joint damage in patients with rheumatoid arthritis. METHODS: TNF-α and MMP-9 in serum and synovial fluid from patients with early RA and controls were tested with a double-antibody enzyme-linked immunosorbent assay. The correlation between MMP-9 and Larsen score over the first 12 months was analyzed. THP-1 cells differentiated by the treatment with TPA were stimulated with increasing concentration of TNF-α for 24 h in vitro. The protein expression of MMP-9 was determined by Western blotting. The activity of MMP-9 was measured by gelatinolytic zymography. Boyden chamber-matrigel in vitro invasion assay was used to detect the invasive capacity. RESULTS: The levels of TNF-α and MMP-9 in serum and synovial fluid of RA patients were significantly higher than those in controls (P<0.05). Serum and synovial fluid levels of MMP-9 correlated significantly with Larsen score (r=0.37 and 0.32, P<0.01). The MMP-9 activity and invasive ability of co-cultured THP-1 cells with TNF-α and TPA were higher than those of non-TNF-α treatment. CONCLUSION: TNF-α upregulates MMP-9 activation and promotes infiltration of monocyte-derived macrophages, indicating that TNF-α play an important role in the pathogenesis of RA. 相似文献
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AIM:To investigate the role of apolipoprotein E(ApoE) in cholesterol efflux mediated by ATP-binding cassette transporter A1(ABCA1) and ATP-binding cassette transporter G1(ABCG1). METHODS:RAW 264.7 cells were seeded in either 6-well or 24-well plates, and then incubated with 20 mg/L low-density lipoprotein receptor gene knockout(LDLr-/-) mouse lipoprotein 20 mg/L ApoE gene knockout(ApoE-/-) mouse lipoprotein or culture medium alone. The changes of intracellular lipid content were measured by transmission electron microscopy and enzymatic colorimetric method. The cholesterol efflux was determined by liquid scintillation. The mRNA and protein levels of ABCA1 and ABCG1 were detected by real-time PCR and Western blotting, respectively. RESULTS:The ApoE-/- mouse lipoprotein increased the content of intracellular cholesterol ester by 60% compared with the control cells. In addition, ApoE-/- mouse lipoprotein treatment decreased the cholesterol efflux to apolipoprotein A-I(ApoA-I) and high-density lipoprotein(HDL) compared with LDLr-/- mouse lipoprotein treatment. ApoE-/- mouse lipoprotein treatment inhibited the mRNA and protein levels of ABCA1 and ABCG1 compared with LDLr-/- mouse lipoprotein treatment. CONCLUSION:Apolipoprotein E plays an important role in the cholesterol efflux of macrophages, which is associated with its regulatory effect on the expression of ABCA1 and ABCG1. 相似文献
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AIM:To study the action of ATP binding cassette transporter(ABC) A 1 on cholesterol efflux in THP-1 macrophage-derived foam cells.METHODS:After exposure of the cultured THP-1 macrophage-derived foam cells to 22(R)-hydroxycholesterol and 4, 4'-diisothiocyanostilbene-2, 2'-disulfonic acid (DIDS) at different concentration for 24 hours, cholesterol efflux and ABCA1 mRNA level were determined by FJ-2107P type liquid scintillator and reverse trancriptase-polymerase chaim reaction(RT-PCR), respectively.RESULTS:Oxidized LDL promoted cholesterol efflux in THP-1 macrophages and 22(R)-hydroxycholesterol increased cholesterol efflux in THP-1 macrophage-derived foam cells in a dose-dependent manner and DIDS inhibited cholesterol efflux in THP-1 macrophage-derived foam cells in a dose-dependent manner. Exposure of the cultured THP-1 macrophage-derived foam cells to 22(R)-hydroxycholesterol and DIDS at different concentration for 24 hours, resulted in increase and decrease in the expression of ABCA1 mRNA in THP-1 macrophage-derived foam cells in a dose-dependent manner, respectively.CONCLUSION:ABCA1 playes an important role in cholesterol efflux in THP-1 macrophage-derived foam cells. 相似文献
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AIM: To investigate the regulation of ghrelin on the expression of ATP-binding cassette transporter A1 and G1 (ABCA1/ABCG1)during the foam cell formation. METHODS: The human monocytic leukemia cell line (THP-1)was chosen in our study. The differentiation of THP-1 cells into macrophages was induced by using phorbol myristate acetate (PMA). Macrophages were then incubated with oxidized LDL (ox-LDL)to generate foam cells. Ghrelin of different concentrations were treated at different time points during foam cell formation. The ABCA1/ABCG1 protein and mRNA levels were detected by Western blotting and RT-PCR. The effect of variance of cholesterol content was measured by zymochemistry via-fluorospectrophotometer. RESULTS: Ghrelin reduced the content of lipid droplet in foam cells, and increased the efflux of intracellular cholesterol significantly. Ghrelin increased ABCA1 protein mass and mRNA level in dose-dependent manner. The changes of ABCG1 protein and mRNA level were the same as ABCA1. CONCLUSION: Ghrelin interfere atherosclerosis by up-regulating the expression of ABCA1 and ABCG1. 相似文献
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MA Wang-ge ZHOU Dong WANG Li-jun LIU Yan CHEN Xiao-li GUO Ning ZHOU Juan 《园艺学报》2018,34(12):2127-2131
AIM:To investigate the effects of interleukin-17A (IL-17A) on the expression of adenosine triphosphate binding cassette transporter A1 (ABCA1) in RAW264.7 macrophages. METHODS:Mouse RAW264.7 macrophages were treated with IL-17A at different concentrations for 6 or 24 h, or treated with IL-17A at the same concentration for different time. The expression of ABCA1 at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. Cholesterol efflux to apolipoprotein A1 (ApoA-1) was evaluated by NBD-cholesterol method. Lipid accumulation in the cells was evaluated by Oil Red O staining. RESULTS:Compared with control group, IL-17A increased the expression of ABCA1 at protein level in the RAW264.7 cells significantly (P<0.05), but had no effect on the mRNA expression of ABCA1. In addition, cholesterol efflux to ApoA-1 was increased and lipid accumulation in the RAW264.7 cells was decreased obviously after treatment with IL-17A. CONCLUSION:IL-17A increases the protein expression of ABCA1 but not at mRNA level in the RAW264.7 macrophages, which may be correlated with its anti-atherosclerosis effect. 相似文献
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AIM: To investigate the perturbative effects of inflammatory stress on cholesterol efflux in human kidney mesangial cells (HMCs) and the relation to peroxisome proliferators activated receptor-γ (PPARγ)-1iver X activated receptor-α (LXRα)-and ATP-binding cassette transporter A1 (ABCA1) pathway. METHODS: HMCs were cultured and divided into control group (incubated with serum free medium), high lipid group , inflammatory stress group or combination treatment group . The mRNA and protein levels of PPARγ, LXRα,ABCA1 were examined by real-time polymerase chain reaction (PCR) and Western blotting. cholesterol assay was performed to evaluate the efflux of cholesterol by liquid scintillation counter. Oil red O staining was used to evaluate lipid droplet accumulation in the cells. Intracellular cholesterol level was measured by enzymic assay. RESULTS: : LDL increased the expression of PPARγ, LXRα and ABCA1 at mRNA and protein levels in HMCs, while TNF-α reduced the expression of these genes at mRNA and protein levels. The cholesterol efflux was increased after LDL loading. However, inflammatory stress inhibited cholesterol efflux in the absence or presence of LDL loading. Oil red O staining and quantitative analysis showed that LDL loading increased the intracellular cholesterol level in HMCs and inflammatory stress further exacerbated the lipid accumulation. CONCLUSION: Inflammatory cytokine reduces cholesterol efflux by inhibiting the expression of PPARγ, LXRα and ABCA1, thereby causing lipid accumulation in HMCs. 相似文献
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YANG Zhi-hong GONG Wei CHEN Feng-ling ZHOU Wen-bai ZHANG Shuo LI Lian-xi LIANG Wen-chang YANG Ye-hong HU Ren-ming 《园艺学报》2008,24(10):2029-2032
AIM: To study the effect of astragalus polysaccharides (Aps) on cholesterol efflux in THP-1 macrophage-derived foam cells. METHODS: After exposed to Aps at different doses, cholesterol efflux and ABCA1 protein levels in cultured THP-1 macrophage-derived foam cells were determined by a γ counter and flow cytometry, respectively. RESULTS: Aps increased cholesterol efflux in THP-1 macrophage-derived foam cells with dose dependent pattern and resulted in an increase in the expression of ABCA1 protein in THP-1 macrophage-derived foam cells. CONCLUSION: The increase in cholesterol efflux by Aps might be related to the up-regulation of ABCA1. 相似文献
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AIM:To study whether astragaloside affects the expression of ATP binding cassette transporter A1 (ABCA1) by regulating miR-33a and promotes the outflow of cholesterol in macrophages. METHODS:In the in vivo experiments, HE staining was used to detect the pathological damage of the cross section of aorta in the mice. The expression of ABCA1 at mRNA and protein levels in mouse aorta was determined by real-time PCR and Western blot. In the in vitro experiments, THP-1 macrophage-derived foam cells were established and then treated with astragaloside-containing serum. Real-time PCR was used to detect the expression of miR-33a. The cells were randomly divided into blank serum group, astragaloside serum group and astragaloside serum+miR-33a mimic group. The expression of ABCA1 at mRNA and protein levels was determined by real-time PCR and Western blot. Oil red O staining and high-performance liquid chromatography were used to detect intracellular lipid content. The method of[3H] incorporation was used to detect intracellular cholesterol outflow. RESULTS:In vivo experiments showed that the blood vessels of the mice in astragaloside group were structurally normal, with neat arrangement, localized small calcified particles, mild lesions, small plaques, reduced foam cells and li-pid, and basically complete elastic plates, indicating that the pathological changes were significantly lighter than those in model group. Compared with model group, the expression of miR-33a in the aorta of the mice in astragaloside group was decreased and the relative expression of ABCA1 at mRNA and protein levels was increased (P<0.05). In vitro experiments showed that astragaloside significantly up-regulated the expression of ABCA1 at mRNA and protein levels, but this effect was inhibited by the transfection of miR-33 mimic without affecting the cell viability. Astragaloside reduced the lipid accumulation in the cells, but this effect was attenuated by miR-33 mimic. Astragaloside reduced intracellular cholesterol accumulation in relation to its promotion of intracellular cholesterol efflux, and the transfection of miR-33a mimic in the cells inhibited cholesterol efflux. CONCLUSION:Astragaloside inhibits the production of miR-33a to increase the expression of ABCA1 and promote the outflow of cholesterol in macrophages. This may be one of the molecular mechanisms of astragaloside in preventing atherosclerosis. 相似文献
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AIM:To investigate the effect of cholesterol metabolite 27-hydroxycholesterol (27-OHC) on the proliferation of lung cancer cells. METHODS:Human lung cancer A549 cells were treated with 27-OHC at different concentrations (0, 0.3125, 0.625, 1.25, 2.5, 5 and 10 μmol/L) for 24~48 h. The cell viability, cell cycle, cell prolife-ration, the intracellular cholesterol levels and cholesterol metabolism-related molecule expression were subsequently assessed by CCK-8 assay, flow cytometry, EdU staining, tissue total cholesterol detection kit, real-time PCR and Western blot. RESULTS:27-OHC decreased the viability of the A549 cells in a dose-and time-dependent manner (P<0.01) and inhibited the cell proliferation (P<0.05). The expression of typical liver X receptor (LXR) downstream target proteins including ATP-binding cassette transporter A1 (ABCA1), low-density lipoprotein receptor (LDLR), and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CR) were modulated, which promoted the efflux of intracellular cholesterol, and reduced cholesterol influx and de novo synthesis, resulting in decreased intracellular cholesterol levels and cell viability. Furthermore, the inhibitory effect of 27-OHC on A549 cell viability was significantly attenuated after the LXR pathway was partially blocked by 5 μmol/L GSK2033 treatment (P<0.05). CONCLUSION:27-OHC inhibits A549 cell prolife-ration via activation of LXR signaling pathway. 相似文献
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AIM: To study the protective effect of puerarin on the atherosclerosis of RAW264.7-derived foam cells. METHODS: The model of foam cells was established by incubating the RAW264.7 cells with ox-LDL. The cholesterol uptake was evaluated by a DiI-ox-LDL binding assay. The ability of cholesterol efflux of the RAW264.7-derived foam cells was detected by cholesterol efflux assay. The protein levels of LC3Ⅱ, P62, CD36, ABCA1, LAL and p-AMPK were determined by Western blot. RESULTS: Puerarin treatment reduced the cholesterol uptake capacity and enhanced the cholesterol efflux rate. The protein levels of LC3Ⅱ, ABCA1 and LAL in puerarin group were higher than that in ox-LDL group, while the protein levels of P62 and CD36 were obviously decreased, and those in rapamycin treatment group had the same change as puerarin group. The protein levels of LC3Ⅱ, ABCA1 and LAL were obviously decreased and the protein level of p-AMPK was increased after co-treated with 3-MA. CONCLUSION: Puerarin promotes LAL and ABCA1-mediated cholesterol efflux in ox-LDL-treated RAW264.7 macrophages, which might enhance autophagy through AMPK-dependent pathway for cholesterol efflux regulation, and reduce the uptake of lipids by CD36 negative regulation. 相似文献
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SHAO Bo LI Ying WANG Xiao-yan ZHAI Xiao-tian TIAN Hua LIU Bo-yan ZHANG Ying SI Yan-hong QIN Shu-cun 《园艺学报》2000,36(10):1754-1760
AIM To verify whether Cordyceps militaris polysaccharide （CMPS） has the effect of promoting reverse cholesterol transport （RCT） in vitro and in vivo , and to explore the underlying mechanism. METHODS For in vivo experiments, RCT efficiency was detected in cholesterol ester transporter transgene （CETP-tg） mice by isotope tracer technique, and the plasma lipid levels were measured by enzyme method. For in vitro experiments, the residual lipid content after cholesterol efflux in RAW264.7 macrophage-derived foam cells was tested by oil red O staining and total cholesterol （TC） kit. Western blot was used to analyze the protein expression of the molecules involved in cholesterol transport, uptake and transformation in the foam cells and mice liver. RESULTS After 4 weeks of intragastric administration of CMPS, the concentrations of TC, low-density lipoprotein cholesterol （LDL-C） and high-density lipoprotein cholesterol （HDL-C） in the plasma of CETP-tg mice were reduced by 24%, 23% and 22%, respectively. RCT efficiency of CETP-tg mice was accelerated and the appearance of 3H-cholesterol tracer in plasma, bile, intestine and feces was significantly increased in CMPS group. Meanwhile, the expression levels of cholesterol receptors scavenger receptor B1 （SR-B1） and LDL receptor （LDLR）, and cholesterol converting rate-limiting enzyme cholesterol 7α-hydroxylase A1 （CYP7A1） were upregulated by 105%, 71% and 58% in the liver of CMPS group, respectively. The results of in vitro experiments showed that CMPS preincubation promoted cholesterol efflux, decreased intracellular lipid and TC levels, and up-regulated the expression of peroxisome proliferator-activated receptor γ （PPARγ）-liver X receptor α （LXRα）-ATP-binding cassette transporter A1 （ABCA1）/ABCG1 signaling pathway related proteins in macrophage-derived foam cells. CONCLUSION CMPS promotes excess cholesterol efflux from peripheral macrophage-derived foam cells and accelerates its discharge through liver pathway. PPARγ-LXRα-ABCA1/ABCG1 pathway may be involved in the mechanism. 相似文献
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AIMTo investigate the role of soluble Klotho protein in THP-1-derived foam cell formation. METHODSTHP-1 monocytes were induced into macrophages by treatment with 160 nmol/L phorbol myristate acetate for 48 h, and then were divided into 6 groups: negative control group (THP-1-derived macrophages), positive control group [THP-1-derived foam cells induced by oxidized low-density lipoprotein (ox-LDL) for 48 h], and 25, 50, 100 and 200 μg/L soluble Klotho protein groups (THP-1-derived macrophages pretreated with soluble Klotho protein at the indicat?ed concentraions for 2 h and then induced by ox-LDL for 48 h). Lipid droplets in cytoplasm were observed by oil red O staining. The cholesterol outflow rate was detected by scintillation counting technique. The content of intracellular total cholesterol, free cholesterol and cholesterol ester was detected by enzyme fluorescence analysis. The expression of acyl-coenzyme A:cholesterol acyltransferase 1 （ACAT1） and ATP-binding cassette transport?er A1 (ABCA1) at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. RESULTSOil red O staining and lipid mass quantification showed that THP-1-derived foam cell formation was dose-dependently suppressed by soluble Klotho protein. The cholesterol efflux rate of THP-1-derived foam cells was increased by soluble Klotho protein in a dose-dependent manner (P <0.05). In addition, soluble Klotho protein decreased the expression of ACAT1 and increased the expression of ABCA1 in a dose-dependent manner (P <0.05). CONCLUSION The soluble Klotho protein inhibits THP-1-derived foam cell formation in a dose-dependent manner by down-regulating the expression of ACAT1 and up-regulating the expression of ABCA1. 相似文献
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AIM To explore the anti-atherosclerotic mechanism of Wendan decoction based on formation of foam cells. METHODS The optimal concentrations of Wendan decoction without cytotoxity to cells were selected by MTT assay. After Wendan decoction treatment, the formation of foam cells was examined by oil red O staining. The cholesterol efflux, cholesterol level, free cholesterol level and cholesterol esterification rate were analyzed using cholesterol efflux assay, total cholesterol assay and free cholesterol assay. The expression levels of macrophage membrane proteins, including CD36, scavenger receptor class A （SR-A）, ATP-binding cassette transporter A1 （ABCA1） and scavenger receptor class B type I （SR-BI）, were quantified by Western blot. RESULTS The optimal concentrations of Wendan decoction without cytotoxity to the cells were 0~6 g/L. Wendan decoction at the concentrations of 1.5, 3 and 6 g/L were selected for the experiments. Wendan decoction at these concentrations inhibited the formation of foam cells induced by oxidized low-density lipoprotein （ox-LDL）, and reduced the accumulation of intracellular lipids in a concentration-dependent manner （P <0.05 or P <0.01）. Wendan decoction also reduced intracellular total cholesterol level, cholesterol ester level and cholesterol esterification rate （P <0.05 or P <0.01）, promoted efflux of intracellular cholesterol （P <0.01）, and decreased the protein level of CD36 in THP-1 cell-derived macrophages （P <0.01） in a concentration-dependent manner. Wendan decoction at the concentration of 6 g/L significantly reduced the protein level of SR-A in THP-1 cell-derived macrophages （P <0.05）. At the concentrations of 3 and 6 g/L, Wendan decoction significantly increased the protein levels of ABCA1 and SR-BI in THP-1 cell-derived macrophages （P <0.05 or P <0.01）. CONCLUSION Wendan decoction significantly inhibits ox-LDL-induced formation of foam cells by reducing cholesterol deposition and promoting cholesterol efflux, and its mechanism may be related to the down-regulation of CD36 and SR-A and the up-regulation of ABCA1 and SR-BI. 相似文献
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YU Si-yang WANG Yan ZENG Gao-feng LIU Yang XU Jian-qiang ZENG Meng-ya TANG Ye-hua ZENG Zhi-ying SHI Xiao-qiao CHEN Ying ZHAO Guo-jun 《园艺学报》2016,32(5):863-868
AIM: To explore the role of nucleotide-binding oligomerization domain-like receptor protein 1 (NLRP1) inflammasome in atorvastatin-induced reduction of interleukin-1β (IL-1β) and interleukin-18 (IL-18) releases from the THP-1 macrophages. METHODS: Lipopolysaccharide (LPS, 10 μg/L) was used to trigger the secretion of IL-1β and IL-18 in the THP-1 macrophages. The cells were incubated with different concentrations of atorvastatin (1, 10 and 20 μmol/L) for 24 h, or treated with 10 μmol/L atorvastatin for different time (12 h, 24 h and 48 h). NLRP1 siRNA was transfected into the THP-1 cells. The mRNA expression of NLRP1 inflammasome was detected by RT-PCR. The protein expression of NLRP1 inflammasome was determined by Western blot. The secretion of proinflammatory cytokines IL-1β and IL-18 was quantified by ELISA. RESULTS: Atorvastatin inhibited the mRNA and protein expression of NLRP1 inflammasome in the THP-1 macrophages in a dose- and time-dependent manner. Transfection of NLRP1 siRNA significantly decreased the protein expression of NLRP1 and promoted the suppressive effect of atorvastatin on IL-1β and IL-18 secretion in the THP-1 macrophages. CONCLUSION: Atorvastatin inhibits the production of IL-1β and IL-18 in the macrophages through decreasing NLRP1 inflammasome expression, possibly contributing to the anti-inflammatory effect of atorvastatin on atherosclerosis. 相似文献