同型半胱氨酸诱导内皮细胞凋亡的研究进展

近年的研究发现，血浆同型半胱氨酸(homocysteine，HCY)水平升高是心肌梗死、脑梗死等心脑血管疾病以及外周阻塞性血管疾病的一个独立危险因素。动物实验发现，HCY可以通过损伤血管内皮细胞，启动粥样斑块和血栓形成。在粥样斑块内存在明显的内皮细胞凋亡。体外内皮细胞培养也显示HCY能诱导其凋亡，提示HCY可能通过内皮细胞凋亡而促进动脉粥样硬化。目前关于HCY引起内皮细胞凋亡的机制尚不明确，作者就HCY的代谢、HCY与内皮细胞凋亡的关系以及细胞凋亡的产生机制综述如下。     

上调SIRT1减少同型半胱氨酸诱导的血管内皮细胞凋亡

目的 研究同型半胱氨酸处理对血管内皮细胞中SIRT1表达的影响,并探讨上调血管内皮细胞中SIRT1表达对同型半胱氨酸诱导的血管内皮细胞凋亡的影响.方法 建立同型半胱氨酸诱导的血管内皮细胞凋亡模型,通过CCK8试剂盒检测细胞活性,通过流式细胞仪检测细胞凋亡,通过qPCR法检测mRNA表达和通过免疫印记法检测蛋白表达.结果...     

葛根素对同型半胱氨酸损伤内皮细胞的影响

目的探讨葛根素对同型半胱氨酸损伤血管内皮细胞的保护作用。方法建立同型半胱氨酸（homocysteine，Hey）诱导的人脐静脉内皮细胞株（ECV304细胞）损伤模型，通过四甲基偶氮唑盐比色法（MTT）检测不同浓度鹞根素对ECV304细胞增殖的影响。结果HCY明显抑制ECV304细胞增殖，葛根素浓度为10-500μ/ml时，细胞活力较模捌组有明显改善；其中当葛根素浓度为100μg／ml时细胞活力最好（P〈0．001）；但与正常组比较仍有显著性差异（P〈0．001）。当葛根索浓度为20mg／ml时，细胞活力较模型组低。结论葛根索在一定浓度范围对内皮细胞有保护作用。     

miR-221通过细胞周期蛋白D1介导同型半胱氨酸诱导人冠状动脉内皮细胞损伤

目的 研究miR-221通过细胞周期蛋白D1介导同型半胱氨酸(Hcy)诱导人冠状动脉内皮细胞(HCAEC)损伤.方法 培养HCAEC并分为4组,对照组用无血清培养基处理,Hcy组用含有1 mmol/LHcy的培养基处理,Hcy+NC(阴性对照)组转染NC抑制物后用含有1 mmol/L的培养基处理,Hcy+miR-221...     

同型半胱氨酸对内皮细胞及脂质过氧化的影响

为观察同型半胱氨酸对内皮细胞,内皮细胞一氧化氮系统及脂质过氧化的影响,在培养兔主动脉内皮细胞中加入兔天然低密度脂蛋白后分为七组;对照组不加任何药物,叶酸组;四氢喋呤ＢＨ４）组,一氧化氮供体药ＳＩＮ－１组,同型半胱氨酸组;同型半胱氨酸加叶酸组和同型半胱氨酸加四氢喋呤组。     

同型半胱氨酸与冠状动脉病变支数关系的探讨

目的探讨血浆同型半胱氨酸(Homocysteine,Hcy)水平与冠状动脉病变支数的关系.方法94例按冠状动脉造影结果分为冠状动脉狭窄(CAS)组(72例)和冠状动脉正常(CAN)组(22例),并根据冠状动脉病变程度进一步将CAS组分为1支病变组(32例),2支病变组(27例),3支病变组(13例),其中非重度狭窄者(38例),重度狭窄者(34例).用高效液相色谱法测定血浆Hcy水平.结果CAS组血浆Hcy水平显著高于CAN组(P＜0.002).0、1、2、3支血管病变者血浆Hcy浓度呈逐级增高趋势,分别为(8.25±3.57)μmol/L、(11.19±4.87)μmol/L、(12.00±5.77)μmol/L、(15.20±5.29)μmol/L(P＜0.01),多重比较3、2、1支与0支及3支与1支病变组,差异有显著性(P＜0.05～0.01),1支与 2支及2支与3支病变组比较差异无显著性(P＞0.05).重度狭窄者血浆Hcy水平显著高于非重度狭窄者,差异有显著性(P＜0.02).结论Hcy水平升高与冠心病密切相关,冠状动脉病变支数越多,血浆Hcy水平越高.     

血清同型半胱氨酸水平与冠状动脉狭窄程度的相关性研究

目的探讨血清同型半胱氨酸(Hcy)水平与冠状动脉狭窄程度的相关性。方法选取2013-04~2014-12该院心内科收治的84例疑似冠状动脉硬化性心脏病(CHD)患者为研究对象,均行冠脉造影检查和Hcy检测,分析CHD与血清Hcy之间的关系。结果非CHD组Hcy水平均低于不同程度冠状动脉狭窄组,差异有统计学意义(P0.05),Hcy水平随着冠状动脉狭窄程度的升高而升高,差异均有统计学意义(P0.05)。血清Hcy水平与冠状动脉狭窄程度积分呈正相关(r=0.835),差异有统计学意义(P0.05)。结论血清Hcy水平与冠状动脉狭窄程度呈正相关,可作为临床预测CHD严重程度及预后的一个重要指标。     

同型半胱氨酸诱导内皮细胞凋亡及舒降之的拮抗机制

目的：探讨同型半胱氨酸（Hcy）是否通过caspase途径诱导内皮细胞凋亡,以及辛伐他汀是否可以通过调节内源性凋亡抑制蛋白c—IAP发挥其拮抗效应。方法：Hcy、辛伐他汀或两者联合处理人脐静脉内皮细胞（HUVEC）24h后,用An—nexinV染色加流式细胞技术及末端脱氧核苷酸转移酶原位标记技术了解细胞凋亡状态;RT—PCR和蛋白质印迹技术检测caspase3、c—IAP1和c—IAP2的mRNA和蛋白质水平。结果：0．5mmol／L和3mmol／L的Hcy均可导致HUVEC凋亡,Hcy浓度高时凋亡细胞数较多,并伴有caspase3表达和活化增强,以及c—IAP1和c—IAP2的mRNA和蛋白质水平降低,辛伐他汀可以上调c—IAP1和c—IAP2的表达。结论：Hcy诱导HUVEC凋亡,此种作用可能涉及caspase3相关途径。辛伐他汀可促进C—IAP系统的表达,从而部分桔抗Hcy的作用。     

同型半胱氨酸诱导内皮细胞凋亡及舒降之的拮抗机制

目的:探讨同型半胱氨酸(Hcy)是否通过caspase途径诱导内皮细胞凋亡,以及辛伐他汀是否可以通过调节内源性凋亡抑制蛋白c-IAP发挥其拮抗效应。方法:Hcy、辛伐他汀或两者联合处理人脐静脉内皮细胞(HUVEC)24h后,用An-nexinV染色加流式细胞技术及末端脱氧核苷酸转移酶原位标记技术了解细胞凋亡状态;RT-PCR和蛋白质印迹技术检测caspase3、c-IAP1和c-IAP2的mRNA和蛋白质水平。结果:0.5mmol/L和3mmol/L的Hcy均可导致HUVEC凋亡,Hcy浓度高时凋亡细胞数较多,并伴有caspase3表达和活化增强,以及c-IAP1和c-IAP2的mRNA和蛋白质水平降低,辛伐他汀可以上调c-IAP1和c-IAP2的表达。结论:Hcy诱导HUVEC凋亡,此种作用可能涉及caspase3相关途径。辛伐他汀可促进c-IAP系统的表达,从而部分拮抗Hcy的作用。     

川芎嗪对脂多糖诱导人冠状动脉内皮细胞炎症损伤的保护作用及机制

目的观察川芎嗪(LIG)对脂多糖(LPS)诱导的人冠状动脉内皮细胞(HCAEC)炎症损伤的影响并探讨其可能的机制。方法用1、10、100μmol/L LIG预处理HCAEC 12 h,再加入1 mg/L LPS共同处理12 h。HCAEC的细胞活力通过噻唑蓝法检测; HCAEC的凋亡率通过流式细胞术检测; HCAEC细胞培养液上清中白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的水平通过酶联免疫吸附法检测; HCAEC细胞中三磷酸腺苷结合盒转运体A1(ABCA1)蛋白的表达通过Western blot法检测。结果 LPS组细胞的活力较对照组显著降低,HCAEC细胞凋亡率和培养液上清中IL-6和TNF-α的水平较对照组显著增加,细胞中ABCA1蛋白表达较对照组显著下调(均P0. 05)。LIG(10、100μmol/L)+LPS组细胞活力较LPS组显著增加,细胞凋亡率和培养液上清中IL-6和TNF-α的水平较LPS组显著降低,细胞中ABCA1蛋白表达较LPS组显著上调(均P0. 05)。结论 LIG抑制LPS诱导的HCAEC的炎症损伤,其机制可能与LIG上调ABCA1的表达有关。     

The association of homocysteine and coronary artery disease

Hyperhomocysteinemia has been associated with increased risk of atherosclerosis and myocardial infarction by a number of prospective case-control studies. A variety of genetic mutations, nutritional deficiencies, disease states, and drugs can elevate homocysteine concentrations. Treatment with folic acid with or without B-complex vitamins effectively lowers homocysteine levels. Whether therapy corresponds with decreased risk of coronary events is unknown, but may be promising. This article reviews the biochemistry of homocysteine metabolism, pathogeneisis, and etiology of hyperhomocysteinemia, along with its association with coronary artery disease, screening, and treatment.     

内皮脂肪酶检测在冠状动脉粥样硬化性心脏病患者中的意义

目的结合血清高敏C反应蛋白（high—sensitivityC—reactionprotein,hs．CRP）,探讨血清内皮脂肪酶（endotheliallipase,EL）与冠状动脉粥样硬化性心脏病（冠心病）及冠状动脉病变程度的关系。方法对入选对象行冠状动脉造影,同步进行心电图、外周血EL、hs—CRP、肌酸激酶同工酶、心肌肌钙蛋白I参数检测和常规身体检查后设正常对照组（30例）和冠心病组（85例）;再将冠心病组分为稳定型心绞痛亚组（39例）、急性冠脉综合征亚组（46例）;同时也将85例冠心病患者分为冠状动脉单支病变（34例）亚组与多支病变（51例）亚组。比较组间外周血EL、hs．CRP浓度差异,评价两者在冠心病患者中的相关性及其与冠状动脉病变程度的相关性。结果冠心病组血清EL、hs—CRP浓度显著高于正常对照组,差异有统计学意义[EL：（762．91±72．62）μg／Lvs（368．23＋42．14）μg／L,P〈0．05：hs—CRP：（12．76±3．08）mg／Lvs（2．34±1．53）mg／L,P〈0．05]。急性冠脉综合征组血清EL及hs．CRP浓度显著高于稳定型心绞痛组,差异有统计学意义[（856．44±36．85）μg／Lvs（478．26±24．53）μg／L,P〈0．05;（21．89±4．36）mg／Lvs（5．76±6．21）mg／L,P〈0．05]。冠心病组中,51例冠状动脉多支病变患者血清EL浓度明显高于34例单支病变患者,差异有统计学意义[（817．98±51．53）μg／Lvs（705．42±34．61）μg／L,P〈0．05]。所有患者血清EL浓度与hs．CRP浓度呈正相关（r=0．327,P=0．02）。结论EL在预测冠心病中有重要价值,它们在冠心病外周血中浓度的升高可以提示冠状动脉粥样斑块的不稳定,且在预测冠状动脉病变程度方面也有一定作用。     

Insulin and its analogue glargine do not affect viability and proliferation of human coronary artery endothelial and smooth muscle cells

Aims/hypothesis Present guidelines for the treatment of type 2 diabetes recommend HbA₁c values of less than 7%. As beta cell function worsens during progress of the disease, insulin therapy is often necessary to achieve this ambitious goal. However, due to peripheral insulin resistance, many patients need rather high insulin dosages. In the light of the extremely high cardiovascular risk of diabetic patients, it is important to determine whether high concentrations of insulin or its frequently used analogues are harmful to the cardiovascular system. We therefore investigated the modulatory effects of regular human insulin and its analogue glargine on proliferation and apoptosis of human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs).Methods Cells were treated with regular human insulin or insulin glargine. Proliferation was determined by [³H]thymidine incorporation and by flow cytometric analysis of Ki-67 expression. Apoptosis was assessed by flow cytometry (cell cycle analysis and annexin V staining) and determination of caspase-3 activity.Results HCAECs and HCASMCs treated with regular human insulin or insulin glargine did not show significant increases in DNA synthesis or Ki-67 expression. Administration of regular human insulin or insulin glargine did not modulate the extent of apoptotic events. No influence of insulin on lipoapoptotic vascular cell death could be detected.Conclusions/interpretation Taken together, neither regular human insulin nor insulin glargine influences growth and apoptosis of human coronary artery cells in vitro. Our data do not suggest that regular human insulin or insulin glargine promote atherosclerosis through mechanisms affecting the cellularity of human coronary arteries.     

Ghrelin对同型半胱氨酸诱导大鼠心肌微血管内皮细胞损伤和炎性反应的保护作用

目的探讨Ghrelin对同型半胱氨酸(Hcy)诱导大鼠心肌微血管内皮细胞(CMECs)损伤和炎性反应的保护作用及作用机制。方法取雄性SD大鼠心肌细胞培养鉴定后,分别以不同浓度的Hcy、不同浓度Ghrelin孵育24 h,采用MTT法检测细胞活力。另选细胞随机分为对照组、Hcy组(0.25 mmol/L)、Ghrelin组(100 ng/L)和混合组,Hoechst染色计算细胞凋亡率,NO试剂盒检测NO的分泌活性,ELISA检测白细胞介素6(IL-6)、细胞间黏附分子1(ICAM-1)的分泌,免疫印迹法检测NF-κB蛋白的表达。结果不同浓度Hcy细胞活性较不同浓度Ghrelin明显降低(P0.05,P0.01)。与对照组比较,Hcy组NO的分泌活性显著降低,细胞凋亡率显著上升,IL-6和ICAM-1的分泌显著增加,NF-κB的表达显著增加,差异有统计学意义(P0.05,P0.01)。与Hcy组比较,混合组可以明显抑制上述指标(P0.05)。结论 Hcy可诱发大鼠CMECs的损伤和炎性反应,而Ghrelin预处理对Hcy诱导的损伤及炎性反应有明显的抑制作用,其保护作用机制可能与抑制NF-κB信号通路有关。     

Statin therapy in patients with coronary artery disease improves the impaired endothelial progenitor cell differentiation into cardiomyogenic cells

Human endothelial progenitor cells (EPCs) can differentiate into cardiomyogenic cells in vitro. We tested the effects of statin therapy on the differentiation rate of EPCs from patients with coronary artery disease (CAD), who may benefit from autologous cell therapy.EPCs from 3 age-matched groups were tested: No CAD (n = 13), CAD patients with (n = 10) or without (n = 16) statin therapy. From 4 CAD patients, EPCs were tested before and after 4 weeks of therapy with 20 mg atorvastatin. After 6 days of co-culture with rat neonatal cardiomyocytes, EPC differentiation was quantified by immunostaining for -sarcomeric actinin flow cytometry analysis. After 6 days of co-culture, the percentage of -sarcomeric actinin–positive EPCs was significantly (p = 0.014) higher in EPCs from adults without CAD (8.07% ± 1.48% of EPCs) compared to EPCs from CAD patients without statin (3.56% ± 0.72%). Importantly, patients with statin therapy revealed significantly higher numbers of -sarcomeric actinin-positive EPCs (6.36% ± 0.69%, p = 0.01) compared to CAD patients without statin. In addition, statin therapy resulted in a significant (p = 0.017) increase of EPC differentiation in all 4 CAD patients investigated before and 4 weeks after statin therapy. The survival of EPCs did not differ between the different groups suggesting that the regulation of EPC differentiation is not secondary to altered EPC survival. In vitro, EPC treatment with 0.1 µM atorvastatin did not affect EPC differentiation (116.15% ± 49.11% of control).EPCs from patients with CAD display impaired differentiation into cardiomyogenic cells. This defect can be improved by in vivo, statin therapy.     

Impact of left ventricular ejection fraction on endothelial function in patients with coronary artery disease

BACKGROUND: Endothelial dysfunction is present in patients with coronary artery disease (CAD) or with congestive heart failure. HYPOTHESIS: This study was performed to evaluate the impact of systolic heart function on endothelial function in patients with CAD. METHODS: The study population consisted of 283 consecutive patients (mean age 59 years, 176 men) undergoing coronary angiography. Endothelial function was assessed by measuring flow-mediated vasodilation (FMD) of the brachial artery. RESULTS: Patients (n = 236) with an ejection fraction (EF) > or = 55% on routine echocardiogram were younger (mean age 58 vs. 62 years), showed a lower prevalence of diabetes (15 vs. 38%) and myocardial infarction (13 vs. 66%), and showed a higher FMD (4.8 +/- 2.4 vs. 4.0 +/- 2.0%, p < 0.05) than patients (n = 47) with an EF < 55%. The correlation coefficient between FMD/endothelial function and EF/systolic heart function was 0.149 (p < 0.02) in the overall study population. Multivariate analysis showed that of age, gender, frequency of diabetes mellitus, myocardial infarction, and CAD extent, EF was the only significant independent parameter correlating with FMD in patients with CAD. CONCLUSIONS: Compared with the other tested risk factors, EF surprisingly was the only significant independent parameter correlating with endothelial function in patients with CAD. Our results support the view that endothelial function is an independent prognostic factor in patients with CAD.     

冠心病患者外周血内皮祖细胞数量及生物学功能与肝细胞生长因子的关系

目的观察冠心病患者外周血内皮祖细胞（EPCs）数量及生物学功能的变化,进一步探讨肝细胞生长因子（HGF）对其影响,为临床应用HGF提供理论依据。方法收集50例非冠心病患者（对照组）、50例冠心病患者（冠心病组;每例分为HGF干预组和非HGF干预组）外周血,应用流式细胞仪和ELISA法分别检测各组CD133＋／CD34＋细胞的数量和HGF水平;采用密度梯度离心法分离培养各组外周血中EPCs,通过MTT法、Transwell迁移试验、黏附能力测定试验及PI—AnnexinV双重染色法来分别检测EPCs的增殖、迁移、黏附能力和凋亡水平。结果与对照组比较,冠心病组外周血中CD133＋／CD34＋细胞数量减少[（2．15±0．69）％1）S（5．26±1．16）％,P〈0．011,血浆中HGF浓度升高[（6．80±1．22）w（2．62±0．83）gg／L,P〈0．01],EPCs增殖、迁移、黏附等生物学功能减弱（P〈0．05）;HGF干预组EPCs增殖、迁移、黏附等生物学功能显著改善（P〈0．05）。各组细胞凋亡水平差异无统计学意义（P〉0．05）。结论外周血中CD133＋／CD34＋细胞数量和血浆中HGF水平的变化可能成为冠心病患者新的危险评估因素。