血凝素样氧化低密度脂蛋白对单核巨噬细胞源泡沫细胞基质金属蛋白酶-9的调控作用

目的观察氧化低密度脂蛋白(ox-LDL)对人单核巨噬细胞源泡沫细胞分泌的基质金属蛋白酶-9(MMP-9)的表达和活性的影响,以及此过程中血凝素样氧化低密度脂蛋白受体-1(LOX-1)的调控作用。方法体外原代培养人单核细胞来源的巨噬细胞,传至2～6代用于实验。依不同浓度ox-LDL(20、40、80mg/L)作用24h,以及ox-LDL40mg/L作用1、12、24h分组,另设LOX-1受体阻断剂多聚肌苷酸(PolyI)干预组。反转录-聚合酶链反应(RT-PCR)测定LOX-1和MMP-9mRNA表达,酶谱法分析MMP-9的活性。结果 ox-LDL作用后,LOX-1和MMP-9mRNA表达升高,MMP-9的分泌及活性升高(P<0.05),且诱导作用呈浓度-时间依赖性;多聚肌苷酸抑制后,MMP-9mRNA表达及活性明显下降(P<0.05)。结论 LOX-1作为ox-LDL的特异性受体,介导ox-LDL诱导的上调单核巨噬细胞LOX-1、MMP-9表达及活性的作用。     

氧化低密度脂蛋白诱导体外培养的巨噬细胞CRP的表达

目的:证实作为动脉粥样硬化病变中重要的炎细胞成分之一的巨噬细胞可表达CRP,并受氧化低密度脂蛋白(OX-LDL)的调节。方法:以不同浓度的OX-LDL处理外周血单核细胞(PBMC)来源的巨噬细胞和THP-1巨噬细胞,应用酶联免疫吸附实验(ELISA)测定浓缩的上清液中CRP的含量,并应用反转录聚合酶链反应(RT-PCR)测定CRP在mRNA水平上的变化。结果:来源于PBMC的巨噬细胞可表达CRP,其浓度相对较低,为(0.45±0.37)μg/L,OX-LDL以浓度依赖性方式显著增加CRP的生成,100mg/LOX-LDL组CRP增至(8.12±0.43)μg/L,但后加入的LPS可降低相应浓度的OX-LDL所诱导的CRP的表达,50mg/LOX-LDL 1μg/LLPS组与50mg/LOX-LDL组比较,CRP的表达有显著性差异(P<0.01)。以THP-1巨噬细胞为材料的RT-PCR也进一步证实以上结果。结论:巨噬细胞可产生CRP并受OX-LDL的调节,局部生成的CRP直接参与动脉粥样硬化的形成。     

巨噬细胞对低密度脂蛋白的修饰和PGE2抗AS作用的实验研究

目的研究巨噬细胞对低密度脂蛋白(LDL)的修饰和前列腺素E2(PGE2)对动脉粥样硬化形成的抑制作用。方法以巨噬细胞(Mp)作用LDL,并设PGE2(20mg/L)组,检测每组过氧化脂质含量,谷胱甘肽过氧化物酶活性,及形态计量学测定细胞变化和含脂量。结果细胞修饰组的脂质过氧化物含量高于给PGE2组,而谷胱甘肽过氧化物酶活性显著低于给药组。作用24小时的修饰组细胞面积明显增大,摄脂增加,均分别显著高于给药组和细胞对照组。结论提示Mp能够氧化修饰LDL并能摄取脂质,导致泡沫细胞形成。大剂量PGE2有抗LDL氧化修饰,抑制Mp细胞摄脂,和泡沫细胞形成,从而抑制动脉粥样硬化发生的作用。     

2型糖尿病动脉粥样硬化的早期检验方法

2型糖尿病（T2DM）动脉粥样硬化（AS）是多因素所致．除高血压、高血糖外，血脂异常如高低密度脂蛋白、高甘油三酯和低高密度脂蛋白等也是重要的致病原因。近年来大量研究显示氧化低密度脂蛋白（OX—LDL）在动脉粥样硬化发生发展中起着重要作用。本研究目的是探讨OX—LDL在T2DM AS发生发展中的作用，建立一早期诊断T2DM有无AS的方法。     

三百棒多糖的单糖组成分析及对巨噬细胞泡沫化的作用研究

目的 研究三百棒多糖(Toddalia asiatica Lam polysaccharides,TALP)的单糖组成及TALP对巨噬细胞泡沫化形成的影响,初步探讨TALP抗动脉粥样硬化的可能机制.方法 采用水提醇沉法提取多糖,除蛋白后干燥,得粗多糖,利用Sephadex-G200纯化三百棒粗多糖,得TALP组分1(T...     

瑞舒伐他汀联合氯沙坦对氧化型低密度脂蛋白诱导的人单核巨噬细胞肝X受体表达的影响

目的探讨瑞舒伐他汀、氯沙坦联合对氧化低密度脂蛋白(ox-LDL)诱导的人单核-巨噬细胞肝X受体(LXR)表达的影响及可能的机制。方法分离并培养人单核细胞,转化为巨噬细胞。据实验要求设置空白对照组、ox-LDL模型对照组、氯沙坦组、瑞舒伐他汀组、瑞舒伐他汀联合氯沙坦组,反转录聚合酶链反应(RTPCR)法测定各组LXR的mRNA的表达。结果与空白对照组比较,ox-LDL模型对照组LXRmRNA表达明显降低(P<0.05);氯沙坦组与ox-LDL模型对照组比较,LXRmRNA表达显著增加(P<0.05);瑞舒伐他汀组与OX-LDL模型对照组比较,LXR mRNA表达显著增加(P<0.05);瑞舒伐他汀联合氯沙坦组与ox-LDL模型对照组比较,LXR mRNA表达显著增加(P<0.05),且高于单药组(P<0.05)。结论瑞舒伐他汀、氯沙坦均可上调LXR的表达,而瑞舒伐他汀与氯沙坦以适宜剂量联合后,LXR的表达增高比单独用药更为显著。     

苯扎贝特对内皮细胞LOX-1及抗凋亡基因Bcl-2影响

目的研究苯扎贝特对氧化型低密度脂蛋白(ox-LDL)作用的人脐静脉内皮细胞(HUVECs)血凝素氧化型低密度脂蛋白受体-1(LOX-1)及抗凋亡基因Bcl-2的影响。方法体外培养HUVECs,反转录聚合酶链反应(RT-PCR)观察各组LOX-1及Bcl-2 mRNA的变化,Western-blot方法观察各组Bcl-2蛋白的变化。结果ox-LDL组与正常对照组比较LOX-1表达显著增加(P<0.05),抗凋亡基因Bcl-2表达降低(P<0.05),苯扎贝特组LOX-1 mRNA表达减少(P<0.05),Bcl-2 mRNA和蛋白表达增加(P<0.05)且呈浓度效应依赖关系。结论苯扎贝特可通过下调LOX-1的表达,上调抗凋亡基因Bcl-2表达从而抑制ox-LDL引起的内皮细胞凋亡。     

PCSK9抑制剂调脂作用的研究进展

前蛋白转化酶枯草杆菌蛋白酶9型(PCSK9)与血脂代谢关系密切,其通过竞争性结合低密度脂蛋白受体(LDLR),导致低密度脂蛋白(LDL)降解减少,引发以低密度脂蛋白胆固醇(LDL-C)水平升高为特点的脂代谢紊乱.PCSK9抑制剂可通过抑制PCSK9与LDLR结合、抑制PCSK9表达发挥调脂作用,降低LDL-C水平.目前...     

中药抗巨噬细胞泡沫化的研究进展

单核细胞分化的巨噬细胞吞噬大量的氧化低密度脂蛋白（oxLDL）,造成脂质聚集形成泡沫细胞,这一过程称为巨噬细胞泡沫化。巨噬细胞泡沫化是动脉粥样硬化早期病变的关键环节。oxLDL的过度摄取、过多胆固醇酯的聚集导致巨噬细胞中形成脂滴,是巨噬细胞泡沫化的主要机制。本文主要从中药调控胆固醇摄取及胆固醇逆转运的相关机制进行概述,总结近年来中药抗巨噬细胞泡沫化的相关研究进展,为中药抗动脉粥样硬化的进一步研究提供参考。     

疫苗接种预防动脉粥样硬化

多项研究证明,在动脉粥样硬化形成中涉及天然和获得性免疫应答,包括促进和拮抗动脉粥样硬化形成的免疫应答.在动脉粥样硬化实验和人体模型中,已确认了诱导免疫应答的共同自身抗原,如氧化型低密度脂蛋白、β2糖蛋白1和热休克蛋白60.已证明氧化型低密度脂蛋白相关抗原能诱导动脉粥样硬化获得性免疫应答.通过热休克蛋白65/60和β2糖蛋白1激活黏膜免疫来诱导免疫耐受也能预防动脉粥样硬化.最近已鉴定出低密度脂蛋白载脂蛋白B-100上的特异性免疫活性表位,早期实验证明,特异性载脂蛋白B-100相关抗原主动免疫是预防动脉粥样硬化的新型免疫调节策略.     

Effect of IL-10 on formation of foam cell induced by ox-LDL

Atherosclerosis is a chronic disease that causes various cardiovascular complications. It has been realized that cellular and humoral immunity plays crucial roles in atherogenic lesion formation. In this study the effects of lipopolysaccharide (LPS) and interleukin-10 (IL-10) on the formation of foam cells during the early stages of atherosclerosis have been investigated. Macrophage was induced by phorbol myristate acetate (PMA) treatment on THP-1 cells. The cells were further stimulated by ox-LDL, ox-LDL plus LPS, ox-LDL plus IL-10 and LPS. By using an oil red O staining technique, the formation of foam cells was evaluated by lipid granules formation in the cells. The ratio of foam cell formation was increased from (9.77 ± 1.70)% to (16.27 ± 2.27)% after 24 h stimulation with ox-LDL, and the increase was observed with incubating time. The foam cells were significantly increased in the presence of LPS in a dose-dependent manner. The maximum increase of about 40% was observed. However, the significant elevation by LPS was abrogated when IL-10 was added. These results indicated that IL-10 can effectively prevent the formation of foam cells induced by ox-LDL with or without LPS. This study demonstrates that ox-LDL can cause foam cell formation from macrophages in vitro. LPS can significantly accelerate this event. IL-10, an anti-inflammatory cytokine, can inhibit the effect of ox-LDL and LPS. These results indicate that inflammatory effects in blood vessels can speed up foam cell formation. The inhibitive effect of IL-10 is an important factor for delaying atherosclerosis processes. __________ Translated from Journal of Tongji University (Medical Science), 2007, 28(1): 1–5 [译自: 同济大学学报(医学版)]     

Protein expressions in macrophage-derived foam cells：comparative analysis by two-dimensional gel electrophoresis

目的:研究氧化低密度脂蛋白诱导的U937泡沫细胞中蛋白质组的差异表达。方法:泡沫细胞模型由人类单核细胞系白血病U937细胞经氧化低密度脂蛋白诱导而成,将对照U937细胞与U937泡沫细胞的蛋白提取物分别进行双向凝胶电泳(2-DE)分离,凝胶经银染后,于PDquest 2-DE图像分析软件中进行图像处理,利用ExPASy蛋白质组学网站中U937细胞的蛋白表达谱信息,对所得的U937泡沫细胞2-DE图谱作进一步鉴定。结果:通过对2-DE蛋白图谱的分析,共150点与参考胶相匹配(匹配率:75％),与U937细胞图谱相比,泡沫细胞中有37个蛋白点表达量有显著性变化(P<0.05),与对照U937细胞相比,在U937泡沫细胞中28个蛋白点表达量上调,9个点表达量下调,其中,8个U937细胞中表达的蛋白点,在U937泡沫细胞中未检测到;而11个在泡沫细胞中表达的蛋白点,未在对照U937细胞中表达。结论:本研究首次建立经氧化低密度脂蛋白诱导的U937泡沫细胞的蛋白差异表达谱,从而提供了对动脉粥样硬化病理过程中巨噬源性泡沫细胞功能性分析研究的新思路。     

Ginsenoside metabolite IH 901 reduced foam cell formation in vitro

Objective To explore the roles of IH 901 on the formation of foam cell sourced from rat peritoneal macrophage.Methods The cultured rat peritoneal macrophages were incubated with ox-LDL to get foam cell as model group.Different doses of IH 901 were added into the culture medium(10 μM,25 μM,50 μM).Foam cell was stained by oil red O.The content of total cholesterol and cholesterol ester were examined to show the formation of foam cells.And the concentrations of TNF-α and IL-1 in culture supernatant were detected.The expression of NF-κB,perilipin and CD36 were measured by western blotting.Results Compared with model group,the formation of foam cell was significantly reduced by IH 901 treatment(50 μM,25 μM),while the amount of living cells of 50 μM group was lessened compared with other groups.Compared with model group,the contents of total cholesterol,cholesterol ester and CE/TC ratio were significantly reduced in IH 901 50 μM and 25 μM groups(P<0.05).Meanwhile,there was obvious difference among IH 901 groups.The concentrations of TNF-α and IL-1 in IH 901 high and middle dose groups were reduced remarkable compared with model group.The changes of expression of NF-κB,perilipin and CD36 showed the similar tendency of the results above.There were no noticeable difference between model group and IH 901 low dose group.Conclusions IH 901 could reduce the formation of foam cell sourced from rat peritoneal macrophage dose dependently.The mechanism may depend on the effects of IH 901 reducing the inflammation reaction of macrophage and the expression of perilipin.     

Anti-atherosclerotic potential of gossypetin via inhibiting LDL oxidation and foam cell formation

Gossypetin, a flavone originally isolated from Hibiscus species, has been shown to possess antioxidant, antimicrobial, and antimutagenic activities. Here, we investigated the mechanism(s) underlying the anti-atherosclerotic potential of gossypetin. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) scavenging activity assay showed that the addition of > 50 μM of gossypetin could scavenge over 50% of DPPH radicals. The inhibitory effects of gossypetin on the lipid and protein oxidation of LDL were defined by thiobarbituric acid reactive substance (TBARS) assay, the relative electrophoretic mobility (REM) of oxidized LDL (ox-LDL), and fragmentation of apoB in the Cu^2 +-induced oxidation of LDL. Gossypetin showed potential in reducing ox-LDL-induced foam cell formation and intracellular lipid accumulation, and uptake ability of macrophages under non-cytotoxic concentrations. Molecular data showed that these influences of gossypetin might be mediated via peroxisome proliferator-activated receptor α (PPARα)/liver-X receptor α (LXRα)/ATP-binding cassette transporter A1 (ABCA1) and PPARγ/scavenger receptor CD36 pathways, as demonstrated by the transfection of PPARα siRNA or PPARγ expression vector. Our data implied that gossypetin regulated the PPAR signals, which in turn led to stimulation of cholesterol removal from macrophages and delay atherosclerosis. These results suggested that gossypetin potentially could be developed as an anti-atherosclerotic agent.     

巨噬细胞对低密度脂蛋白的修饰和PGE2抗AS作用的实验研究

目的：研究巨噬细胞对低密度脂蛋白（LDL）的修饰和前列腺素E2（PGE2）对动脉粥样硬化形成的抑制作用。方法：以巨噬细胞（Mp）作用LDL,并没PGE2（20mg/L组）,检测每组过氧化脂质含量,谷胱甘肽过氧化物酶活性,及形态计量学测定细胞变化和含脂量。结果：细胞修饰组的脂质过氧化物含量高于给PGE2组,而谷胱甘肽过氧化物酶活性显著低于给药组。作用24小时的修饰组细胞面积明显增大,摄取增加,均分别显著高于给药组和细胞对照组。结论：提示Mp能够氧化修饰LDL并能摄取脂质,导致泡沫细胞形成。大剂量PGE2有抗LDL氧化修饰,抑制Mp细胞摄脂,和泡沫细胞形成,从而抑制动脉样硬化发生的作用。     

不稳定性心绞痛患者血管内皮功能及普罗布考干预研究

目的:观察普罗布考对不稳定性心绞痛患者内皮依赖舒张功能、血氧化低密度脂蛋白(ox-LDL)和高敏C反应蛋白(hs-CRP)水平的影响。方法:将53例不稳定性心绞痛患者随机分为普罗布考组和常规治疗组,治疗前、治疗3个月后应用高频超声测肱动脉内皮依赖性舒张功能(FMD),并观察外周血ox-LDL及hs-CRP等指标变化。结果:治疗后普罗布考组FMD较治疗前及常规治疗组治疗后均明显升高(P<0.01),血浆ox-LDL及hs-CRP水平较治疗前及常规治疗组治疗后均明显降低(P<0.01或P<0.05)。普罗布考组治疗后血浆ox-LDL与FMD呈负相关(r=-0.651,P<0.01)。结论:普罗布考具有较强的抗氧化及抗炎症作用并可改善不稳定心绞痛患者内皮依赖舒张功能。     

Exendin-4 decreases liver inflammation and atherosclerosis development simultaneously by reducing macrophage infiltration

Background and Purpose

The aetiology of inflammation in the liver and vessel wall, leading to non-alcoholic steatohepatitis (NASH) and atherosclerosis, respectively, shares common mechanisms including macrophage infiltration. To treat both disorders simultaneously, it is highly important to tackle the inflammatory status. Exendin-4, a glucagon-like peptide-1 (GLP-1) receptor agonist, reduces hepatic steatosis and has been suggested to reduce atherosclerosis; however, its effects on liver inflammation are underexplored. Here, we tested the hypothesis that exendin-4 reduces inflammation in both the liver and vessel wall, and investigated the common underlying mechanism.

Experimental Approach

Female APOE*3-Leiden.CETP mice, a model with human-like lipoprotein metabolism, were fed a cholesterol-containing Western-type diet for 5 weeks to induce atherosclerosis and subsequently treated for 4 weeks with exendin-4.

Key Results

Exendin-4 modestly improved dyslipidaemia, but markedly decreased atherosclerotic lesion severity and area (−33%), accompanied by a reduction in monocyte adhesion to the vessel wall (−42%) and macrophage content in the plaque (−44%). Furthermore, exendin-4 reduced hepatic lipid content and inflammation as well as hepatic CD68⁺ (−18%) and F4/80⁺ (−25%) macrophage content. This was accompanied by less monocyte recruitment from the circulation as the Mac-1⁺ macrophage content was decreased (−36%). Finally, exendin-4 reduced hepatic chemokine expression in vivo and suppressed oxidized low-density lipoprotein accumulation in peritoneal macrophages in vitro, effects dependent on the GLP-1 receptor.

Conclusions and Implications

Exendin-4 reduces inflammation in both the liver and vessel wall by reducing macrophage recruitment and activation. These data suggest that exendin-4 could be a valuable strategy to treat NASH and atherosclerosis simultaneously.     

脂蛋白对培养巨噬细胞表达噬巨细胞移动抑制因子的实验研究

目的:探讨不同脂蛋白对培养的人巨噬细胞表达巨噬细胞移动抑制因子基因和蛋白水平的影响。方法:2 8SC人巨噬细胞株用含有1×10 5U·L-1青霉素、10 0mg·L-1链霉素和10 ?S的RPMI 16 40培养基于37℃,5 %的CO2 中培养。以每孔1×10 4个细胞接种于6孔板中,每孔2ml培养液,加入终浓度为15 0mg·L-1的不同脂蛋白,于37℃共同孵育2 4h收集细胞和培养介质。采用RT PCR和ELISA分别测定MIFmRNA和蛋白表达水平。结果:巨噬细胞有MIF表达,低密度脂蛋白、氧化型低密度脂蛋白、胆固醇和甘油三脂组诱导巨噬细胞MIFmRNA和蛋白表达水平,试验组较正常对照组升高(P <0 .0 5 ) ;而高密度脂蛋白和极低密度脂蛋白组诱导巨噬细胞MIFmRNA和蛋白表达水平,试验组与正常对照组变化相近(P >0 .0 5 )。结论:不同脂质对巨噬细胞MIFmRNA和蛋白表达作用不同。MIF可能参与了低密度脂蛋白、氧化型低密度脂蛋白、胆固醇和甘油三脂所致的动脉粥样硬化进程。     

Modulation of hepatic and extrahepatic LDL receptors: Involvement in the progression of atherosclerosis

Extensive investigation over the past 2 decades has led to a far better understanding of the role of lipoproteins in the etiology and progression of atherosclerosis. Paramount to this understanding has been the elucidation of how cells recognize these macromolecular complexes and metabolize the lipids and proteins they transport. Because of their elevation in plasma of many individuals with premature coronary heart disease, low-density lipoproteins (LDL) have received particular attention. As primary transporters of cholesterol to cells in mammalian tissues, LDL particles are removed from blood via specific cell surface receptors that facilitate their uptake. Once internalized and exposed to lysosomal enzymes, LDL particles are degraded, “freeing” their cholesterol to regulate cellular cholesterol synthesis and modulate the number of LDL receptors on the cell membrane. This high-affinity, high-capacity process has been termed the LDL receptor pathway and is functional in most tissues of the body, especially in liver where over 50%percnt; of the total body LDL degradation occurs. Liver is the target organ of bile acid sequestrants (cholestyramine/colestipol) and HMG-CoA reductase inhibitors (compactin/mevinolin), both of which induce increased expression of LDL receptors by affecting hepatic cholesterol availability. Unlike liver, the arterial wall under normal conditions is not very active at degrading LDL via the classical LDL receptor pathway. However, when hypercholesterolemia occurs, especially when there is an LDL-receptor deficiency, the arterial wall removes 20--fold more LDL than normal and with a greater fractional catabolic rate. Thus other endocytotic pathways exist by which LDL can enter cells of the arterial wall. These pathways are not as actuely regulated as the classical LDL receptor pathway, and, as a result, the arterial wall cells, particularly smooth muscle cells and resident macrophages, accumulate cholesteryl esters. In this brief review, the importance of these receptors to the etiology and progression of atherosclerosis is discussed in detail and sites of possible pharmacological intervention are identified.