高糖诱发氧化应激导致血管内皮细胞凋亡机制研究

糖尿病是一组由于遗传因素和环境因素共同作用而导致的临床综合征,其血管并发症是导致糖尿病患者致死和致残的主要原因。血管内皮细胞凋亡在糖尿病血管并发症的发生发展过程中起重要作用。高糖能诱发氧化应激,通过一系列复杂的信号通路,如NF-κB信号通路、JNK信号通路、PI3K/AKT信号通路等直接或间接引起血管内皮细胞凋亡。改善高糖诱导的细胞凋亡可能成为治疗糖尿病及其各种并发症的一种有效途径。     

葛根素对缺氧性血管内皮细胞凋亡的保护作用

目的 研究葛根素(puerarin)对缺氧性血管内皮细胞凋亡的影响。方法 用NaCN合并无糖培养基造成培养的牛主动脉内皮细胞(BAECs)缺氧;用台盼蓝染色、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)、流式细胞仪计数和Hoechst 33342荧光染色法观察细胞受损和凋亡情况;用免疫细胞化学染色法观察细胞内Caspase-3的表达情况。结果NaCN合并无糖培养基可引起血管内皮细胞凋亡。葛根素可显著减少缺氧性内皮细胞凋亡,并对Caspase-3的表达有明显抑制作用。结论葛根素对缺氧条件下的血管内皮细胞有保护作用,此作用至少部分通过抑制Caspase-3的表达而实现。     

共轭亚油酸对血管内皮细胞的保护作用

目的探讨多不饱和脂肪酸共轭亚油酸(CLA)对血管内皮细胞(VEC)的保护作用。方法应用MTT比色法测定CLA对血管内皮细胞存活率的影响;用绿色荧光蛋白标记的膜联蛋白/碘化丙啶双染色法检测细胞凋亡和死亡。结果CLA 2μmol·L-1处理细胞72h,细胞存活率为126.3%;在浓度小于5μmol·L-1时呈剂量和时间性依赖。CLA能部分阻断软脂酸(PA)和硬脂酸(SA)诱导的VEC死亡。结论CLA能部分抵抗饱和脂肪酸PA、SA诱导的血管内皮细胞死亡,对血管内皮细胞的存活率有重要意义。     

线粒体凋亡通路在棕榈酸诱导的血管内皮细胞凋亡中的作用

目的探讨线粒体凋亡通路在棕榈酸诱导的人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)凋亡中的作用。方法不同浓度棕榈酸(0.1、0.2、0.4、0.8 mmol·L~(-1))作用HUVECs(0、12、24、48 h),MTT法检测HUVECs增殖能力;免疫荧光法检测HUVECs细胞内活性氧(reactive oxygen species,ROS)的表达水平;免疫印迹法检测HUVECs中AIF、Cyt-C、cleaved caspase-3、Bcl-2、Bax等线粒体凋亡通路蛋白的表达水平;TUNEL法检测细胞内凋亡水平。结果 0.4 mmol·L~(-1)棕榈酸作用HUVECs 24 h时,细胞增殖率明显降低;AIF、Cyt-C、cleaved caspase-3、Bax/Bcl-2的水平明显升高(P<0.05),细胞凋亡水平明显增高(P<0.05);线粒体通透性抑制剂环孢素A (ciclosporine A, CsA)预处理组HUVECs凋亡水平明显下调(P<0.05)。结论棕榈酸所致血管内皮细胞损伤的发病机制可能与线粒体凋亡相关通路密切关联,此研究对脂毒性心肌损伤的防治有重要意义。     

外源性羟自由基诱导血管内皮细胞凋亡及卡托普利的干预作用

目的 观察外源性氧自由基（OFR）衍生的羟自由基（OH^-)诱导培养的VECs凋亡及卡托普利（Captopril,Cap)的干预作用，探讨血管内皮细胞（VECs)凋亡（Apoptosis)与心血管疾病的关系。方法 （1）采用改良的Jaffe法，进行人脐静脉血管内皮细胞（HUVECs)原代、传代培养；（2）取4－6代HUVECs，无血清条件下分别为10^-5、10^-4、10^-3、10^-2、10^-1mol/L羟自由基（OH^-)共孵育，观察细胞存活率、贴壁率、形态学、DNA凝胶电泳、DNA片段率及培养液一氧化氮（NO）含量变化；（3）观察10^-3mol/L OH^-诱导HUVECs凋亡及10^-7、10^-6、10^-5mol/L Cap干预后上述参数的变化。结果 （1）正常培养HUVECs存活率＞95％，贴壁率＞90％；（2）外源性OH－明显降HUVECs存活率、贴壁率，且呈剂量依赖性；10^-5、10^-4、10^-3mol/L OH^-诱导HUVECs凋亡，DNA“梯形”以10^-3mol/L OH^-作用8－24h显著，DNA片段率、凋亡细胞百分率明显升高；而10^-2、10^-1mol/L OH^-可诱导HUVECs坏死；（3）10^-7、10^-6、10^-5mol/L Cap干预明显提高OH^-处理的HUVECs存活率、贴壁率，降低其凋亡细胞百分率、DNA片段率，使DNA“梯形”渐消失，NO分泌减少渐逆转。结论 （1）采用改良的Jaffe法，可以建立HUVECs体外模型；（2）外源性OH^-在较低浓度时可诱导培养HUVECs的典型凋亡；（3）Cap能减轻或抑制OH^-诱导培养HUVECs的凋亡，恢复其NO分泌。     

出血性蛇毒诱导血管内皮细胞凋亡的研究进展

出血性蛇毒中含有多种能专一性诱导血管内皮细胞凋亡的组分,如金属蛋白酶/解离素蛋白家族和L-氨基酸氧化酶。本文介绍了诱导血管内皮细胞凋亡的出血性蛇毒组分以及相关的分子机制。     

牛磺酸与细胞凋亡

牛磺酸(Taurinet)是动物体内的一种含硫氨基酸,它广泛分布于生物体内各组织、器官,主要以游离状态存在于组织间液和细胞内液中,具有广泛的生物学作用。它主要存在于心肌细胞,对心肌细胞、神经细胞、肾脏细胞、平滑肌细胞、肿瘤细胞等细胞的凋亡都有重要的影响,进而能对细胞有一定的保护作用。     

氧化应激在乙醇诱导内皮细胞凋亡中的作用

目的 观察乙醇诱导的内皮细胞凋亡及氧化应激所起的作用.方法 体外培养人内皮细胞株EA.hy926,实验分对照组、实验组（ 0.6% 乙醇）、N-乙酰半胱氨酸（NAC）组（0.6% 乙醇＋ 10 mmol&#8226;L^-1 NAC）,各组以相应药物孵育12 h,采用四甲基偶氮唑蓝法检测细胞抑制率,流式细胞仪检测细胞活性氧（ROS）和凋亡率,相应试剂盒测定超氧化物歧化酶（SOD）活力及丙二醛（MDA）水平.结果与对照组相比,实验组内皮细胞增殖受到明显抑制,SOD活力从(40.8±2.9 )U&#8226;mg^-1蛋白降至（29.2±4.0） U&#8226;mg^-1蛋白、MDA水平由（46.5±3.8） nmol&#8226;mg^-1蛋白升至（89.5±5.6） nmol&#8226;mg^-1蛋白、ROS荧光强度从（93.69±7.58）升高到（162.30±18.85）,凋亡率由（0.49± 0.16）% 增至（5.31±0.54）% (均P＜0.01);而NAC组凋亡率、ROS和MDA水平较实验组均降低（P＜0.01),SOD活力显著恢复.结论 乙醇引起的氧化应激在其诱导的内皮细胞凋亡中起重要作用.     

牛磺酸对原代培养大鼠肝细胞的促凋亡作用研究

目的探索分离培养原代正常大鼠肝细胞的条件和方法,研究牛磺酸对原代培养的正常大鼠肝细胞的促凋亡作用。方法使用经典的两步灌注法分离原代肝实质细胞后贴壁培养,记录细胞生长状况,台盼蓝拒染法测定细胞活率、绘制生长曲线。经用不同浓度的牛磺酸培养24 h后,用TUNEL法测定牛磺酸对原代培养大鼠肝细胞致凋亡作用,测定细胞凋亡率(AI)。结果两步灌注分离取得的肝细胞活率>70%,贴壁生长良好,从生长曲线可看出培养第3天进入对数生长期;TUNEL实验3个给药组AI分别为:3.71%,29.72%,42.56%。结论两步灌注法是一种细胞产量高,损伤小,纯度高的分离原代正常大鼠肝细胞的方法;高浓度的牛磺酸对体外培养肝细胞有一定致凋亡作用。     

牛磺酸对原代培养大鼠肝细胞的促凋亡作用研究

目的探索分离培养原代正常大鼠肝细胞的条件和方法,研究牛磺酸对原代培养的正常大鼠肝细胞的促凋亡作用。方法使用经典的两步灌注法分离原代肝实质细胞后贴壁培养,记录细胞生长状况,台盼蓝拒染法测定细胞活率、绘制生长曲线。经用不同浓度的牛磺酸培养24 h后,用TUNEL法测定牛磺酸对原代培养大鼠肝细胞致凋亡作用,测定细胞凋亡率(AI)。结果两步灌注分离取得的肝细胞活率>70%,贴壁生长良好,从生长曲线可看出培养第3天进入对数生长期;TUNEL实验3个给药组AI分别为:3.71%,29.72%,42.56%。结论两步灌注法是一种细胞产量高,损伤小,纯度高的分离原代正常大鼠肝细胞的方法;高浓度的牛磺酸对体外培养肝细胞有一定致凋亡作用。     

Taurine antagonized oxidative stress injury induced by homocysteine in rat vascular smooth muscle cells

INTRODUCTION Homocysteine is athiol-containing amino acid thatis formed when methionine was converted to cysteine.Plasma homocysteine concentration may increase invarious pathophysiological conditions, including defi-ciency of vitamins such as folic acid, cyanocobalamin,and pyridoxal phosphate etc[1]. Incubation of humanmegakaryocytic cell line DAMI with homocysteine in-hibited respiratory rate and activities of cytochrome coxidase III/ATPase 6, 8 and other respiratory chain en-zymes i…     

Taurine prevents acrylonitrile-induced oxidative stress in rat brain

Acrylonitrile (ACN) is a volatile, toxic liquid used as a monomer in the manufacture of synthetic rubber, styrene plastics, acrylic fiber and adhesives. ACN is a potent neurotoxin and a carcinogen, which produces tumors in rats, particularly gliomas of the brain. A role for free radical-mediated lipid peroxidation in the toxicity of ACN has been suggested. We examined the ability of taurine, an antioxidant amino acid, to attenuate ACN-induced alterations in lipid peroxidation, cellular DNA fragmentation, GSH, vitamin C and vitamin E levels in blood and brain of rats. Rats were administered with ACN at a concentration of 100 ppm in drinking water and sacrificed after 14 and 28 days. The level of lipid peroxidation and the enzymatic and non-enzymatic antioxidants were assayed. The obtained data were compared with those obtained from ACN rats co-treated with taurine for 14 and 28 days. It was observed that taurine treatment counteracted the oxidative stress induced by ACN by reducing the levels of peroxidation, and enhancing the activities of enzymatic and non-enzymatic antioxidants.     

Single-wall carbon nanotubes induce oxidative stress in rat aortic endothelial cells

Oxidative stress is a major factor contributing to endothelial cell damage. Single-wall carbon nanotubes (SWCNTs) have oxidative properties; however, the oxidative effects of SWCNTs on endothelial cells are not fully understood. In the present study, we investigated the effects of oxidative stress induced by SWCNTs on rat aortic endothelial cells (RAECs). Various markers of cellular damage were assessed, such as biochemical and ES immunity indexes, and DNA and protein damage. Our findings suggest that RAEC endured oxidative damage following SWCNT exposure. Specifically, after SWCNTs exposure, non-enzymatic antioxidant glutathione was activated prior to superoxide dismutase activation in order to defend against oxidative stress. Additionally, it was found that as SWCNT concentration increased, so did the stress protein, heme oxygenase-1 (HO-1), expression levels. These changes may induce RAEC damage, and result in many serious diseases.     

Single-wall carbon nanotubes induce oxidative stress in rat aortic endothelial cells

Oxidative stress is a major factor contributing to endothelial cell damage. Single-wall carbon nanotubes (SWCNTs) have oxidative properties; however, the oxidative effects of SWCNTs on endothelial cells are not fully understood. In the present study, we investigated the effects of oxidative stress induced by SWCNTs on rat aortic endothelial cells (RAECs). Various markers of cellular damage were assessed, such as biochemical and ES immunity indexes, and DNA and protein damage. Our findings suggest that RAEC endured oxidative damage following SWCNT exposure. Specifically, after SWCNTs exposure, non-enzymatic antioxidant glutathione was activated prior to superoxide dismutase activation in order to defend against oxidative stress. Additionally, it was found that as SWCNT concentration increased, so did the stress protein, heme oxygenase-1 (HO-1), expression levels. These changes may induce RAEC damage, and result in many serious diseases.     

Taurine reverses endosulfan-induced oxidative stress and apoptosis in adult rat testis

The present study was aimed to investigate the mechanistic aspect of endosulfan toxicity and its protection by taurine in rat testes. Pre-treatment with taurine (100 mg/kg/day) significantly reversed the decrease in testes weight, and the reduction in sperm count, motility, viability and daily sperm production in endosulfan (5 mg/kg/day)-treated rats. Sperm chromatin integrity and epididymal L-carnitine were markedly decreased by endosulfan treatment. Endosulfan significantly decreased the level of serum testosterone and testicular 3β-HSD, 17β-HSD, G6PDH and LDH-X. Sperm Δψm and mitochondrial cytochrome c content were significantly decreased after endosulfan. Testicular caspases-3, -8 and -9 activities were significantly increased but taurine showed significant protection from endosulfan-induced apoptosis. Oxidative stress was induced by endosulfan treatment as evidenced by increased H₂O₂ level and LPO and decreased the antioxidant enzymes SOD, CAT and GPx activities and GSH content. These alterations were effectively prevented by taurine pre-treatment.In conclusion, endosulfan decreases rat testes weight, and inhibits spermatogenesis and steroidogenesis. It induces oxidative stress and apoptosis by possible mechanisms of both mitochondria and non-mitochondria pathways. These data provide insight into the mode of action of endosulfan-induced toxicity and the beneficial role provided by taurine to counteract endosulfan-induced oxidative stress and apoptosis in rat testis.     

血管内皮细胞线粒体氧化应激与动脉粥样硬化

血管内皮细胞损伤是动脉粥样硬化病理过程的起始环节。线粒体氧化应激与血管内皮细胞功能密切相关,线粒体氧化应激通过诱导线粒体自噬、一氧化氮生成减少、炎症反应、细胞代谢失衡和凋亡,导致血管内皮细胞的功能障碍。同时,血管内皮细胞也通过调控线粒体氧化应激维持自身稳态。本文旨在综述动脉粥样硬化病理过程中线粒体氧化应激诱发血管内皮细胞损伤的主要分子信号通路,为后续研究两者间的分子机制提供参考。     

Hydrogen sulfide protects against high-glucose-induced apoptosis in endothelial cells

Hydrogen sulfide (H2S) is the third endogenous gaseous mediator identified after nitric oxide and carbon monoxide. It has been demonstrated that H2S has protective effects on myocardial ischemia/reperfusion-induced cell apoptosis. To date, little is known about the role of H2S in the pathophysiology of diabetic vascular complications. In this study, we investigated the effects of sodium hydrosulfide on high-glucose-induced apoptosis of primary human umbilical vein endothelium cells. Exposure to high glucose (25 mmole/L) for 48 hours resulted in the induction of apoptosis by 41.6% ± 1.01%, which was attenuated by pretreatment with sodium hydrosulfide (50 μmole/L) for 30 minutes. Further investigation of the apoptotic mechanisms in the cells demonstrated that high glucose upregulated the ratio of Bax/Bcl-2 and activated caspase-3 and also increased the levels of reactive oxygen species and malondialdehyde while reducing superoxide dismutase activity. All the above responses could be prevented by pretreatment with 50 μmole/L of sodium hydrosulfide. These results indicated that the protective effects of H2S on endothelial cells in the condition of high glucose might involve an antioxidative stress mechanism.     

Quercetin attenuates vascular calcification by inhibiting oxidative stress and mitochondrial fission

Vascular calcification is a strong independent predictor of increased cardiovascular morbidity and mortality and has a high prevalence among patients with chronic kidney disease. The present study investigated the effects of quercetin on vascular calcification caused by oxidative stress and abnormal mitochondrial dynamics both in vitro and in vivo. Calcifying vascular smooth muscle cells (VSMCs) treated with inorganic phosphate (Pi) exhibited mitochondrial dysfunction, as demonstrated by decreased mitochondrial potential and ATP production. Disruption of mitochondrial structural integrity was also observed in a rat model of adenine-induced aortic calcification. Increased production of reactive oxygen species, enhanced expression and phosphorylation of Drp1, and excessive mitochondrial fragmentation were also observed in Pi-treated VSMCs. These effects were accompanied by mitochondria-dependent apoptotic events, including release of cytochrome c from the mitochondria into the cytosol and subsequent activation of caspase-3. Quercetin was shown to block Pi-induced apoptosis and calcification of VSMCs by inhibiting oxidative stress and decreasing mitochondrial fission by inhibiting the expression and phosphorylation of Drp1. Quercetin also significantly ameliorated adenine-induced aortic calcification in rats. In summary, our findings suggest that quercetin attenuates calcification by reducing apoptosis of VSMCs by blocking oxidative stress and inhibiting mitochondrial fission.     

卡维地洛对过氧化氢致血管内皮细胞氧化应激损伤的保护作用

目的观察卡维地洛(carved ilol)对过氧化氢(hydrogenperoxide,H2O2)致内皮细胞损伤及表面粘附分子表达的影响。方法采用H2O2作为外源性自由基生成系统,模拟内皮细胞的脂质过氧化损伤,建立离体培养的ECV-304细胞氧化应激损伤模型,观察卡维地洛对H2O2致内皮细胞损伤及表面粘附分子表达的影响。结果卡维地洛各浓度组均明显改善H2O2(1.0×10-6mol.L-1)所致ECV-304细胞形态学损伤,提高细胞生存率,降低LDH释放,并可使细胞内及细胞培养液中MDA含量降低,SOD活性升高,亦可下调ICAM-1蛋白及细胞内ICAM-1mRNA表达水平,上述作用随药物浓度增加呈增强趋势。结论卡维地洛可保护内皮细胞结构和功能的完整性,提高内皮细胞抗氧化能力,并从转录水平抑制脂质过氧化诱导的粘附分子表达增加,降低单核-内皮细胞粘附,有利于减少动脉粥样硬化的始动环节和早期事件的发生。     

姜黄素保护血管内皮细胞氧化应激损伤分子机制的研究进展

随着我国经济社会发展,国民生活方式的转变,心血管病的发病人数持续增加,心血管病已成为我国居民的首位死亡原因.冠心病是最常见的心血管病,血管内皮功能障碍是动脉粥样硬化的起因和最早改变,而氧化应激则是导致血管内皮功能不全和心肌损伤的最常见原因.姜黄素是一种提取于草本植物姜黄根茎的天然疏水性多酚,已被证实其可以通过调节基因表达或直接作用在抗肿瘤、抗增殖、抗炎、抗氧化等诸多方面发挥药理作用,该文旨在对姜黄素保护血管内皮细胞氧化应激损伤的最新分子机制作一综述.