天然及氧化修饰脂蛋白对人动脉平滑肌细胞原癌基因…

动脉平滑肌细胞（ＳＭＣ）的增殖在动脉粥样硬化（ＡＳ）的形成过程中极其重要。我们在建立人主动脉ＳＭＣ体外培养方法的基础上,观察了ＬＤＬ,ＶＬＤＬ及ＨＤＬ和相应的氧化修饰型脂蛋白对培养人ＳＭＣ ｓｉｓ,ｊｕｎ,Ｈ－ｒａｓ原癌基因及Ｒｂ抗癌基因转录表达的影响。结果表明：（１）ＨＤＬ对ＳＭＣｓｉｓ,ｊｕｎ,ｒａｓ基因表达无影响;（２）ＬＤＬ和ＶＬＤＬ有使这些基因表达增加的趋势;（３）ｏｘ－ＬＤＬ,ｏｘ－Ｖ     

天然及氧化修饰脂蛋白对人动脉平滑肌细胞原癌基因表达的影响

动脉平滑肌细胞（ＳＭＣ）的增殖在动脉粥样硬化（ＡＳ）的形成过程中极其重要。我们在建立人主动脉ＳＭＣ体外培养方法的基础上,观察了ＬＤＬ,ＶＬＤＬ及ＨＤＬ和相应的氧化修饰型脂蛋白对培养人ＳＭＣｓｉｓ,ｊｕｎ,Ｈ－ｒａｓ原癌基因及Ｒｂ抗癌基因转录表达的影响。结果表明：（１）ＨＤＬ对ＳＭＣｓｉｓ,ｊｕｎ,ｒａｓ基因表达无影响;（２）ＬＤＬ和ＶＬＤＬ有使这些基因表达增加的趋势;（３）ｏｘ－ＬＤＬ,ｏｘ－ＶＬＤＬ和ｏｘ－ＨＤＬ具有使ＳＭＣｓｉｓ,ｊｕｎ,和ｒａｓ基因表达显著增强的作用（Ｐ＜０．０１）,且其作用较相应的天然脂蛋白大（Ｐ＜０．０１）;（４）天然和氧化修饰型脂蛋白对Ｒｂ基因表达均无影响。据上述结果推测：ＬＤＬ,ＶＬＤＬ,ｏｘ－ＬＤＬ,ｏｘ－ＶＬＤＬ和ｏｘ－ＨＤＬ的致ＡＳ作用可能与刺激ＳＭＣｓｉｓ,ｊｕｎ和ｒａｓ原癌基因表达增加有关。     

天然和氧化修饰型LDL,VLDL及HDL对培养人动脉平滑肌细胞原癌…

动脉平滑肌细胞（ＳＭＣ）是动脉粥样硬化（ＡＳ）斑块中的主要细胞,它的增殖在ＡＳ形成过程中极其重要,脂蛋白和氧化修饰型脂蛋白对ＳＭＣ增殖的影响以及ＳＭＣ增殖与原异常表达的关系是当前ＡＳ发病机制研究的热点之一,我们在建立人主动脉ＳＭＣ体外培养方法的基础上,观察了ＬＤＬ,ＶＬＤＬ及ＨＤＬ和相应的氧化修饰型脂蛋白对培养人ＳＭＣｆｏｓ,ｍｙｃ,ｅｒｂ－Ｂ原癌基因转录表达的影响,结果表明：（１）ＨＤＬ对ＳＭＣ     

氧化修饰高密度脂蛋白对培养人动脉平滑肌细胞细胞周期蛋白D_1基因表达的影响

动脉平滑肌细胞（ｓｍ ｏｏｔｈ ｍ ｕｓｃｌｅ ｃｅｌｌ,ＳＭＣ）是动脉粥样硬化（ａｔｈｅｒｏｓｃｌｅｒｏｓｉｓ,ＡＳ）斑块中的主要细胞,它的增殖在ＡＳ形成过程中极其重要．利用体外培养的人主动脉ＳＭＣ,观察了天然高密度脂蛋白（ｎａｔｉｖｅ ｈｉｇｈ ｄｅｎｓｉｔｙ ｌｉｐｏｐｒｏｔｅｉｎ,Ｎ－ＨＤＬ）及氧化修饰ＨＤＬ（ｏｘｉｄｉｚｅｄ ＨＤＬ,ＯＸ－ＨＤＬ）对培养人主动脉ＳＭＣ ｃｙｃｌｉｎ Ｄ１（细胞周期蛋白Ｄ１）基因转录表达的影响．结果表明：（１）Ｎ－ＨＤＬ对ＳＭＣｃｙｃｌｉｎ Ｄ１基因表达无影响（Ｐ＞ ０．０５）;（２）ＯＸ－ＨＤＬ使ＳＭＣｃｙｃｌｉｎ Ｄ１基因表达显著增强（Ｐ＜０．０１）,其表达量随时间（２、１２、２４ ｈ）延长而增加．上述结果表明,ＯＸ－ＨＤＬ的致ＡＳ作用可能与其刺激ＳＭＣｃｙｃｌｉｎ Ｄ１基因表达增加有关．     

氧化修饰使HDL促动脉平滑肌细胞胆固醇流出减少

为了研究氧化修饰对高密度脂蛋白（ＨＤＬ）转运细胞胆固醇地＾３Ｈ－胆固醇负荷的培养人动脉平滑肌细胞（ＳＭＣ）分别与天然ＨＤＬ及Ｃｕ＾２＋ａｋｇ ＨＯＣｌ氧化修饰的ＨＤＬ在３７℃温育不同时间后,分别测定细胞＾３Ｈ－胆固醇清除率。结果发现,温育２４ｈ后,经Ｃｕ＾２＋或ＨＯＬｌ氧修饰后的ＨＤＬ其细胞胆因醇清除率分别较天然ＨＤＬ下降了３０．０％和４３．１％（ｐ〈０．０１）。结果还发现,Ｃｕ＾２＋或ＨＯＣｌ氧     

氧化修饰HDL剌激培养人主动脉平滑肌细胞增殖

平滑肌细胞（smooth muscle cell, SMC）增殖在动脉粥样硬化（atherosclerosis, AS）形成中起着重要作用.氧化修饰HDL（oxidized HDL, OX-HDL）可刺激 ³H-TdR掺入培养人动脉SMC的DNA,促进SMC增殖.以四甲基偶氮唑盐(MTT)法直接观察OX-HDL对培养人动脉SMC增殖细胞数的影响.结果显示,天然HDL（native HDL N-HDL）对SMC增殖没有影响,而OX-HDL则显著刺激SMC增殖（P＜0.01）,N-HDL显著抑制OX-LDL刺激SMC增殖作用（P＜0.01）,而OX-HDL则显著增加OX-LDL刺激SMC增殖作用（P＜0.01）     

氧化修饰HDL刺激培养人主动脉平滑肌细胞增殖

平滑肌细胞（ｓｍｏｏｔｈ ｍｕｓｃｌｅ ｃｅｌｌ,ＳＭＣ）增殖在动脉粥样硬化（ａｔｈｅｒｏｓｃｌｅｒｏｓｉｓ,ＡＳ）形成中起着重要作用。氧化修饰ＨＤＬ（ｏｘｉｄｉｘｅｄ ＨＤＬ,ＯＸ－ＨＤＬ）可刺激＾３Ｈ－ＴｄＲ掺入培养人动脉ＳＭＣ的ＤＮＡ,促进ＳＭＣ增殖。以四甲工偶氮唑盐９ＭＴＴ）法直接观察ＯＸ－ＨＤＬ对培养人动脉ＳＭＣ增殖细胞数的影响。结果显示,天然ＨＤＬ（ｎａｔｉｖｅ ＨＤＬ Ｎ－ＨＤＬ）对     

天然和氧化修饰型LDL、VLDL及HDL对培养人动脉平滑肌细胞原癌基因fos及myc表达的影响

动脉平滑肌细胞（ＳＭＣ）是动脉粥样硬化（ＡＳ）斑块中的主要细胞,它的增殖在ＡＳ形成过程中极其重要。脂蛋白和氧化修饰型脂蛋白对ＳＭＣ增殖的影响以及ＳＭＣ增殖与原癌基因异常表达的关系是当前ＡＳ发病机制研究的热点之一。我们在建立人主动脉ＳＭＣ体外培养方法的基础上,观察了ＬＤＬ,ＶＬＤＬ及ＨＤＬ和相应的氧化修饰型脂蛋白对培养人ＳＭＣｆｏｓ,ｍｙｃ,ｅｒｂ－Ｂ原癌基因转录表达的影响。结果表明：①ＨＤＬ对ＳＭＣｆｏｓ,ｍｙｃ基因表达无影响;②ＬＤＬ和ＶＬＤＬ有使这些基因表达增加的趋势,但与对照比较差异不显著（Ｐ＞０．０５）;③ＯＸ－ＶＬＤＬ,ＯＸ－ＶＬＤＬ和ＯＸ－ＨＤＬ有使ＳＭＣｆｏｓ,ｍｙｃ基因表达显著增强的作用（Ｐ＜０．０１）,且其作用较相应的天然脂蛋白大（Ｐ＜０．０１）．上述结果说明：ＬＤＬ,ＶＬＤＬ,ＯＸ－ＬＤＬ,ＯＸ－ＶＬＤＬ和０Ｘ－ＨＤＬ的致ＡＳ作用可能与刺激ＳＭＣｆｏｓ和ｍｙｃ癌基因表达增加有关。     

血小板生长因子(PDGF-BB)刺激体外培养兔肺动脉平滑肌细胞DNA和蛋白质合成

应用细胞培养、３Ｈ－ＴｄＲ和３Ｈ－Ｌｅｕｃｉｎｅ掺入方法,观察血小板生长因子ＢＢ（Ｐｌａｔｅｌｅｔ－ｄｅｒｉｖｅｄＧｒｏｗｔｈＦａｃｔｏｒＢＢ）对体外培养兔肺动脉平滑肌细胞ＤＮＡ和蛋白质合成的影响。结果表明：（１）当ＰＤＧＦ－ＢＢ浓度为１０ｎｇ／ｍｌ时,３Ｈ－ＴｄＲ掺入值已较对照组显著增高（６２６２．５±４１２．９ｖｓ８３３．５±１２４．０,Ｐ＜０．０５）;当ＰＤＧＦ－ＢＢ浓度为２０ｎｇ／ｍｌ时,３Ｈ－Ｌｅｕｃｉｎｅ掺入值亦较对照线显著增高（１０２１２．８±６３８．３ｖｓ７３４０．３±１１９７．９,Ｐ＜０．０５）。（２）ＰＤＧＦ－ＢＢ浓度在５－２５ｎｇ／ｍｌ范围内,３Ｈ－ＴｄＲ,３Ｈ－Ｌｅｕｃｉｎｅ掺入值与剂量直线相关（ｒＤＮＡ＝０．９７,ｒｐｒｏｔ＝０．９０Ｐ＜０．０５）。说明ＰＤＧＦ－ＢＢ刺激体外培养兔肺动脉平滑肌细胞ＤＮＡ和蛋白质合成。     

胰岛素及cAMP对培养人动脉平滑肌细胞HDL受体功能的影响

对胰岛素ｃＡＭＰ对培养人动脉平滑肌细胞（ＳＭＣ）ＨＤＬ受体功能的影响进行了研究,结果发现：胰岛素使ＳＭＣＨＤＬ受体的结合容量Ｂｍａｘ即受体数目显著下降,而对ＳＭＣＨＤＬ受体的Ｋｄ值亲和力无影响;ｃＡＭｐ则ＳＭＣＨＤＬ受体亲和力增加,而对受体数目无影响。     

Ox-LDL和天然LDL诱导人动脉平滑肌细胞增殖中蛋白激酶A作用的初步研究

汪浩川等研究表明一定量Ｏｘ－ＬＤＬ能刺激培养人动脉ＳＭＣ细胞的增殖［１］,Ｄｅｊａｇｅｒ等采用交叉抑制实验证明兔ＳＭＣ细胞膜上有能结合Ｏｘ－ＬＤＬ的清道夫受体［２］,因此Ｏｘ－ＬＤＬ诱导培养人ＳＭＣ细胞增殖可能是Ｏｘ－ＬＤＬ作用于ＳＭＣ膜清道夫受体后...     

人动脉平滑肌细胞HDL受体酶联测定法

利用辣根过氧化物酶（HRP）与纯化的高密度脂蛋白（HDL）交联,并将人动脉平滑肌细胞（SMC）固定于酶标板上,成功地建立了人动脉平滑肌细胞HDL受体的酶联测定法．1材料与方法1．1材料辣根过氧化物酶（HRP,RZ=3.0,Sigma）;96孔酶标板（Sigma）;培养基DMEM（GIBCO）．1．2人动脉平滑肌细胞培养培养按本室汪浩川[1]方法体外培养．1．3去apoE-HDL3制备人血清去apoE-HDL3（d=1.20～1.175）按改进的磷钨酸钠-氯化镁沉淀密度梯度超速离心法制备[2]．然后按张林华等[3]一次性密度梯度超速离心分离法分离HDL．SDS-PAGE电泳鉴…     

Modulation of DNA synthesis in aortic smooth muscle cells by dinitrotoluenes

Studies were conducted to determine if in vivo exposure to dinitrotoluenes (DNT), which is associated with circulatory disorders of atherosclerotic etiology in humans, is associated with alterations of vascular smooth muscle cells (SMC) consistent with the atherogenic process. Sprague-Dawley rats (150-180 g) were injected IP for 5 days/week for 8 weeks with 2,4- or 2,6-DNT (0.5, 5, or 10 mg/kg) or medium chain triglyceride (MCT) oil. Histopathologic evaluation of aortae from animals exposed to either isomer showed dysplasia and rearrangement of SMC at all doses tested. Reduced ³H-thymidine incorporation was observed in primary cultures of aortic SMC from DNT-exposed animals relative to vehicle controls. This inhibitory response was maintained for up to two passages in culture after which a significant increase in thymidine incorporation was observed. Exposure of SMC from naive animals to DNT in vitro (1–100 µM) did not alter the extent of thymidine incorporation in cycling or growth-arrested cultures. In contrast, exposure to 2,4- or 2,6-diaminotoluene (DAT) (1–100 µM), carcinogens which share toxic metabolic intermediates in common with DNT, inhibited replicative DNA synthesis and stimulated unscheduled DNA synthesis in cycling and growth-arrested cultures of SMC, respectively. Our results suggest that modulation of DNA synthesis in aortic SMC by DNT metabolites generated in vivo contribute to the development of vascular lesions.Abbreviation DAT diaminotuluene - tDNT technical grade dinitrotoluene - DNT dinitrotoluenes - HU hydroxyurea - IP intraperitoneal - LDH lactate dehydrogenase - MCT oil medium chain triglyceride - NPTC non-protein thiol content - RDS replicative DNA synthesis - SEM standard error of the mean - SMC smooth muscle cells - UDS unscheduled DNA synthesis     

人主动脉壁硫酸乙酰肝素蛋白聚糖对培养的人主动脉平滑机细胞合成蛋白聚糖的影响

从人正常胸主动脉分离硫酸乙酰肝素蛋白聚糖（ＨＳＰＧ）,观察其对体外培养的人主动脉平滑肌细胞（ＨＡＳＭＣ）合成ＰＧ的影响。ＨＡＳＭＣ在不加（对照）或加ＨＳＰＧ（１９μｇ醛酸／ｍｌ）的￣（３５）Ｓ－硫酸钠培养液中培养,以标记ＰＧ。继之,培养液及细胞层的４ｍｏｌ／Ｌ盐酸胍提取液中的ＰＧｓ经离子交换及凝胶过滤柱层析分离,发现加ＨＳＰＧ后,培养液中的ＨＳＰＧ,硫酸软骨素ＰＧ（ＣＳＰＧ）及硫酸皮肤素－硫酸软骨素ＰＧ（ＤＳＣＳＰＧ）均明显增高,而细胞层中仅ＨＳＰＧ和ＣＳＰＧ增高,且加ＨＳＰＧ后细胞层的ＤＳＣＳＰＧ分子大小有所不同,进一步分析ＤＳＣＳＰＧ中ＤＳ及ＣＳ含量发现加ＨＳＰＧ组ＨＡＳＭＣ细胞层中的ＤＳ％含量略低于对照组。结果提示ＨＳＰＧ可刺激ＨＡＳＭＣ的ＰＧ合成,其可能与血管壁修复及动脉壁脂质沉积有关。     

氧化修饰HDL对培养人主动脉平滑肌细胞胆固醇流出的影响

大量研究显示,高密度脂蛋白(highdensitylipoprotein,HDL)具有抗动脉粥样硬化(atherosclerosis,AS)作用.这是由于HDL能够促进外周组织如血管壁内皮细胞、平滑肌细胞(smoothmusclecell,SMC)及巨噬细胞储集的胆固醇流出,并将其转移到肝脏通过胆汁分泌而排出体外[1],这一过程...     

In vitro interactions of phenformin with serum lipoproteins and consequences thereof.

Turbidity developed when phenformin was added to human serum; this turbidity increased in a sigmoidal fashion with rising concentrations of phenformin (5–50 nmole/1). Centrifugation produced clearing of the solution, with collection of particulate matter on the surface of the sera.Extraction of control, and phenformin-treated sera with petroleum ether for 15 min. revealed that cholesterol and triglyceride were responsible for the turbidity. Different sera produced different turbidities with a given concentration of phenformin. No significant simple correlation existed between turbidity and serum cholesterol and/or triglyceride levels. The turbidities, produced by the addition of a constant concentration of phenformin to a series of diluted serum samples, were linearly related to the amount of serum present.The turbidities acquired by purified very-low density (VLDL), low-density (LDL), and high-density lipoprotein (HDL) fractions with phenformin were additive, and the turbidity of phenformin-treated serum was accounted for by these lipoprotein fractions. Serum free of lipoproteins did not become turbid when exposed to phenformin. Phenformin added to serum which had previously been delipidated, failed to produce turbidity. The turbidity produced by phenformin was reversible, because it could easily be cleared by dialysis.No significant differences in quantitative immunochemical reactivities were observed when control serum was compared with the subnatant of phenformin-treated serum, as determined by single radial immunodiffusion with LDL antibodies.These in vitro observations may be related to the in vivo hypolipidemic action of phenformin on hyperlipidemic subjects.