血小板衍生生长因子—BB对血管内皮细胞、平滑肌细胞及成纤维细胞增殖的影响

目的 观察血小板衍生生长因子—BB对培养的人血管内皮细胞、兔平滑肌细胞和人成纤维细胞增殖的影响。方法 采用培养的人脐静脉血管内皮细胞、兔动脉血管平滑肌细胞和人血管成纤维细胞，应用^3H—TdR掺入方法，观察血小板衍生生长因子—BB对三种细胞DNA合成的影响。结果 血小板衍生生长因子—BB可促进处于静止状态的三种细胞DNA的合成，并呈现出明显的浓度依赖关系，在30ng／ml的浓度时成纤维细胞DNA的合成达到高峰，在40ng／ml的浓度时内皮细胞、平滑肌细胞DNA的合成达到高峰。成纤维细胞、平滑肌细胞分别在PDCF-BB作用24h和36h年到DNA合成的高峰，内皮细胞在48h时DNA合成量最高。结论 血小板衍生生长因子—BB可明显促进培养的人脐静脉血管内皮细胞、兔动脉血管平滑肌细胞和人血管成纤维细胞的增殖。     

Platelets and Granulocytes, in Particular the Neutrophils, Form Important Compartments for Circulating Vascular Endothelial Growth Factor

The measurement of circulating vascular endothelial growth factor (VEGF) levels as a prognostic factor will gain increasing relevance in the diagnosis and evaluation of treatment in cancer patients. Angiogenesis is an absolute requirement in tumour growth and metastatic disease. In the present study data are presented which indicate that circulating VEGF mainly resides in peripheral blood cells. In 15 healthy volunteers we demonstrated that approximately 34% of the circulating VEGF resides in platelets and approximately 11% in patients with cancer ( n = 4). An important part namely 58% in healthy volunteers and 69% in patients with cancer of the total circulating VEGF is contained in granulocytes, particular in the neutrophils, as confirmed by fluorescence-activated cell sorting (FACS). Also an increased VEGF level per granulocyte is found in patients with cancer (77 microg VEGF/l) compared with the healthy volunteers (164 microg VEGF/l). In contrast only 2% was present in plasma. The biological significance of platelet- or granulocyte-derived VEGF is not yet known. Liberation of VEGF from these compartments could well be of importance for tumour angiogenesis. Therefore, future studies on the clinical value of circulating VEGF as a prognostic factor in cancer patients should include measurements of VEGF in peripheral blood cells.     

Role of Vascular Endothelial Growth Factor in Pulmonary Endothelial Cell Injury by Exercise Challenge in Asthmatic Patients

This study was designed to examine the role of vascular endothelial growth factor (VEGF) in pulmonary endothelial cell injury by exercise in asthmatics. Post-exercise circulating thrombomodulin (TM) levels were significantly increased in asthmatics. Moreover, the increase in TM levels with exercise was significantly correlated with VEGF level in induced sputum from asthmatics (r = 0.80, p = 0.0007). After inhaled steroid therapy, post-exercise TM levels were significantly decreased, but the increase in TM levels with exercise was also correlated with VEGF level (r = 0.60, p = 0.01). Thus, pulmonary endothelial cells stimulated by VEGF in asthmatic airways may be sensitive to exercise challenge.     

Role of Vascular Endothelial Growth Factor in Pulmonary Endothelial Cell Injury by Exercise Challenge in Asthmatic Patients

This study was designed to examine the role of vascular endothelial growth factor (VEGF) in pulmonary endothelial cell injury by exercise in asthmatics. Post-exercise circulating thrombomodulin (TM) levels were significantly increased in asthmatics. Moreover, the increase in TM levels with exercise was significantly correlated with VEGF level in induced sputum from asthmatics (r = 0.80, p = 0.0007). After inhaled steroid therapy, post-exercise TM levels were significantly decreased, but the increase in TM levels with exercise was also correlated with VEGF level (r = 0.60, p = 0.01). Thus, pulmonary endothelial cells stimulated by VEGF in asthmatic airways may be sensitive to exercise challenge.     

软脉化斑汤对高血压合并动脉粥样硬化大鼠血管内皮细胞的保护作用及机制研究

目的研究软脉化斑汤对高血压合并动脉粥样硬化大鼠血管内皮细胞的保护作用及相关机制。方法选取8周龄雄性自发性高血压大鼠20只,随机分为模型组和软脉化斑汤组,每组10只。另选取8周龄WKY雄性大鼠10只作为对照组。检测不同时间点大鼠血压变化,苏木素-伊红(HE)染色分析血管组织病理形态;测定大鼠血清三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平,酶联免疫吸附试验(ELISA)法测定白介素-6(IL-6)、超敏C反应蛋白(hs-CRP)和肿瘤坏死因子-α(TNF-α)水平;同时测定超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化能力(T-AOC);免疫印迹法(Western Blot)测定血管内皮细胞血管细胞黏附分子-1(VCAM-1)及G蛋白信号调节因子16(RGS16)表达水平。结果干预4周、8周软脉化斑汤组大鼠血压低于模型组(P<0.05);与模型组比较,软脉化斑汤组大鼠TG、TC、LDL-C水平降低(P<0.05),HDL-C水平升高(P<0.05),IL-6、hs-CRP、TNF-α水平降低(P<0.05);与模型组比较,软脉化斑汤组大鼠SOD水平、T-AOC均升高(P<0.05),MDA水平下降(P<0.05);与模型组比较,软脉化斑汤组大鼠血管内皮细胞VCAM-1、RGS16水平下降(P<0.05)。结论软脉化斑汤可有效改善动脉粥样硬化大鼠血压及血脂代谢水平,保护血管内皮细胞损伤,其机制可能与抑制炎症反应、氧化应激及下调VCAM-1、RGS16蛋白表达有关。     

血管内皮细胞生长因子与类风湿关节炎发病机制及其治疗的相关性

血管内皮细胞生长因子(VEGF)及其受体以调控类风湿关节炎血管形成为最主要的特点。他对滑膜炎症的发展及滑膜血管翳的形成具有十分重要的作用。其在类风湿关节炎患者滑液和血清中的表达,与疾病严重程度呈相关性。因此,以VEGF及其受体为靶点,将成为治疗类风湿关节炎的新途径。     

保利尔胶囊对冠心病患者血管内皮功能和C-反应蛋白水平的影响

目的 观察保利尔胶囊对冠心病患者血管内皮功能和C-反应蛋白(CRP)的影响.方法 选择冠心病患者87例,随机分为治疗组和对照组,对照组采用常规西药治疗;治疗组在常规西药治疗的基础上加用保利尔胶囊4粒/次,3次/日,服用1个月.观察两组治疗前后血管内皮依赖性舒张功能(EDD)、血浆内皮素-1(ET-1)、一氧化氮(NO)和CRP水平等指标的变化.结果 治疗后,治疗组ET-1均明显下降(P＜0.05),NO明显升高(P＜0.05),但治疗组ET-1、NO较对照组变化更明显(P＜0.05).治疗后的肱动脉EDD,治疗组为(9.67±3.10)％较对照组(4.32±2.78)％明显改善(P＜0.05);而肱动脉内皮非依赖性血管舒张功能(NMD)也得到明显改善(P＜0.05).结论 保利尔胶囊可使血浆ET-1水平明显下降,NO水平明显升高,EDD、NMD明显改善,CRP明显降低,提示保利尔胶囊有显著保护血管内皮功能、抑制炎症和改善临床症状的作用.     

天香丹对冠心病病人血管内皮生长因子及相关因子的影响

目的探讨天香丹对稳定型心绞痛病人血清血管内皮生长因子(VEGF)、一氧化氮(NO)以及一氧化氮合酶(NOS)含量的影响。方法99例冠心病稳定型心绞痛病人随机分为3组,治疗组36例,在西医基础治疗上加天香丹18g/d;对照Ⅰ组34例,在西医基础治疗上加通心络胶囊每日12粒,对照Ⅱ组29例,西医基础治疗基础上加硝酸酯类,28d为1个疗程,观察病人治疗前后心绞痛发作频率和持续时间、心电图的变化,并观察血清VEGF,NO以及NOS含量的变化。结果治疗组与对照Ⅱ组心绞痛发作次数比较,差异有统计学意义(P<0.05)。治疗组、对照Ⅰ组和对照Ⅱ组的心电图总有效率分别为83.33%,70.59%和68.97%,差异无统计学意义(P>0.05)。治疗组治疗后血清NO和VEGF含量与对照Ⅱ组比较,差异均有统计学意义(P<0.01)。结论天香丹可提高血清VEGF,NO以及NO的含量,促进冠状动脉侧支循环的建立,改善心肌缺氧状态,从而达到治疗冠心病的目的。     

麝香保心丸对H<,2>O<,2>诱导人脐静脉内皮细胞增殖及氧化应激的影响

目的 麝香保心丸时H<,2>O<,2>诱导人脐静脉内皮细胞(HUVEC)增殖活性、氧化因子丙二醛(MDA)、超氧化物歧化酶(SOD)及烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶亚基gp91、p22 mRNA表达的影响.方法 体外胶原酶消化法培养人脐静脉内皮细胞;溴脱氧尿嘧啶核苷(BrdU)插入法测定细胞增殖活性;RT-PCR法测定NADPH氧化酶亚基gp91、p22 mRNA的表达;比色法检测细胞上清液MDA、SOD含量.结果 与对照组相比,H<,2>O<,2>组细胞增殖活性明显降低(P<0.01).与对照组相比,1 g麝香保心丸药物血清组细胞增殖活性明显增高(P<0.01);与对照组相比,H<,2>O<,2>组人脐静脉内皮细胞上清液MDA含量显著升高,SOD活性明显降低(P<0.01),而麝香保心丸药物血清呈浓度依赖性抑制H<,2>O<,2>诱导HUVEC细胞上清液MDA水平升高及SOD活性降低,1 g药物血清组上清液MDA含量显著低于H<,2>O<,2>组(P<0.01),SOD活性明显高于H<,2>O<,2>组(P<0.05);RT-PCR结果显示:与对照组相比,H<,2>O<,2>组p22 mRNA表达水平明显升高,对照血清组p22 mRNA表达进一步增加;与对照血清组相比,麝香保心丸1 g、2 g药物血清组p22 mRNA的表达显著降低;麝香保心丸药物血清对H<,2>O<,2>诱导HUVEC细胞NADPH氧化酶gp91 mRNA表达无显著影响.结论 麝香保心丸保护血管内皮细胞可能与抑制H<,2>O<,2>诱导人脐静脉内皮细胞氧化应激有关.     

全蝎纯化液对凝血酶诱导血管内皮细胞释放NO＼TFPI及表达TF的影响

目的探讨全蝎纯化液对凝血酶诱导血管内皮细胞释放一氧化氮（N0）、组织因子途径抑制物（TFPI）及表达组织因子（TF）的影响。方法对传代培养的人脐静脉细胞（HUVEC）（2代～3代）分别给予不同的处理,取上清液测定N0、TFPI,取细胞冻融液测TF活性。结果与空白对照组比较,凝血酶组能明显抑制血管内皮细胞释放TFPI（P〈0．01）,与凝血酶纽比较,全蝎纯化液各剂量组能拮抗TFPI的降低（P〈0．05）。凝血酶促进血管内皮细胞表达TF,较对照组明显增高（P〈0．01）,全蝎纯化各剂量组较凝血酶组则明显降低（P〈0．01）。全蝎纯化液各剂量组促进血管内皮细胞释放N0,较对照组、凝血酶组明显增高（P〈0．01）。结论全蝎纯化液能抑制凝血酶诱导血管内皮细胞表达TF,可拮抗凝血酶抑制TFPI释放,促进血管内皮细胞释放N0,提示全蝎纯化液对凝血酶诱导血管内皮细胞的损伤具有保护作用。     

Autocrine Role of Interleukin-8 in Induction of Endothelial Cell Proliferation,Survival, Migration and MMP-2 Production and Angiogenesis

Interleukin-8 (IL-8/CXCL8), a paracrine angiogenic factor, modulates multiple biologic functions in CXCR1 and CXCR2 expressing endothelial cells. Several reports suggest that inflammation, infection, cellular stress and tumor presence regulate IL-8 production in endothelial cells. In the present study, we test the hypothesis that IL-8 regulates multiple biological effects in endothelial cells in an autocrine manner. We examined the autocrine role of IL-8 in regulating angiogenesis by using a neutralizing antibody to IL-8, CXCR1 or CXCR2 in human vein umbilical endothelial cell (HUVEC) and human dermal microvascular endothelial cell (HMEC). Neutralizing antibody to IL-8, CXCR1 or CXCR2 inhibited endothelial cell proliferation, and MMP-2 production as compared to cells cultured with medium alone or control antibody. In addition, we observed that the number of apoptotic cells was significantly higher in anti-IL-8, anti-CXCR1 and anti-CXCR2 treated endothelial cells, which coincided with decreased survival-associated gene expression. We observed reduced migration of endothelial cells treated with anti-IL-8 and anti-CXCR2 antibody, but not anti-CXCR1 antibody as compared to controls. Further, we observed an inhibition of capillary tube formation and neovascularization following treatment with anti-IL-8, anti-CXCR1 and anti-CXCR2 antibodies. Together these data suggest that IL-8 functions as an important autocrine growth and angiogenic factor in regulating multiple biological activities in endothelial cells.     

Differences in the Effect of Angiotensin-converting Enzyme Inhibitors on the Rate of Endothelial Cell Apoptosis: In Vitro and In Vivo Studies

Aim An imbalance between endothelial apoptosis and regeneration is one of the initiating events in atherosclerosis. Angiotensin-converting enzyme (ACE) inhibition corrects the endothelial dysfunction observed in coronary artery disease, and this could be the consequence of a reduction in the rate of endothelial apoptosis. The aim of this study was to investigate the effect of different ACE inhibitors on endothelial apoptosis. Methods We examined the effect of five ACE inhibitors (enalapril, perindopril, quinapril, ramipril, and trandolapril) on the rate of endothelial apoptosis, either in vitro in human umbilical vein endothelial cells (HUVECs), using a serum deprivation method to induce apoptosis, or in vivo in rats, inducing apoptosis via endotoxic shock with Escherichia coli lipopolysaccharides (LPS). Results We were unable to detect any significant effect of ACE inhibition on the rate of in vitro endothelial apoptosis at concentrations ranging from 5 × 10⁻⁸ to 10⁻⁶ M. In contrast, chronic in vivo administration of ACE inhibitors to rats at dosages that had similar hypotensive effects reduced the rate of LPS-induced apoptosis significantly for perindopril (P < 0.001) and nonsignificantly for the other ACE inhibitors. The order of potency of the ACE inhibitors tested was perindopril > ramipril ≫ quinapril = trandolapril = enalapril, with significant differences between perindopril and quinapril (P < 0.01), trandolapril (P < 0.001), and enalapril (P < 0.001). The difference between perindopril and ramipril did not reach significance. Conclusion Our experiments suggest differences between ACE inhibitors in terms of inhibition of endothelial apoptosis in vivo.     

养心通脉方含药血清预处理内皮祖细胞对急性心肌梗死兔心肌bFGF、VEGF水平的影响

目的观察养心通脉方含药血清预处理内皮祖细胞(EPCs)对急性心肌梗死(AMI)兔心肌血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)水平的影响。方法从35只雄性SPF级兔中随机选取25只建立急性心肌梗死兔模型,随机分为模型组、中药EPCs心肌注射组、中药EPCs耳缘静脉注射组、EPCs心肌注射组、EPCs耳缘静脉注射组,每组5只,另设正常组和假手术组,各5只,通过移植干预后14 d取梗死心肌区及缺血心肌区组织,荧光定量聚合酶链式反应(PCR)检测bFGF、VEGF的mRNA表达,蛋白质印迹法(Western Blot)检测bFGF、VEGF蛋白表达。结果在梗死心肌区及缺血心肌区,中药EPCs心肌注射组和中药EPCs耳缘静脉注射组bFGF、VEGF的mRNA及蛋白表达明显高于假手术组、模型组、EPCs心肌注射组、EPCs耳缘静脉注射组,差异均有统计学意义(P<0.05),但中药EPCs心肌注射组bFGF、VEGF的mRNA及蛋白表达与中药EPCs耳缘静脉注射组比较差异均无统计学意义(P>0.05),EPCs心肌注射组bFGF、VEGF的mRNA及蛋白表达与EPCs耳缘静脉注射组比较差异均无统计学意义(P>0.05)。结论养心通脉方含药血清预处理EPCs,能够明显增加损伤心肌bFGF、VEGF表达,但耳缘静脉注射与心肌注射疗效相当。     

Two cycles of the PS-341/bortezomib, adriamycin, and dexamethasone combination followed by autologous hematopoietic cell transplantation in newly diagnosed multiple myeloma patients

Increased bone marrow angiogenesis is seen in several hematological malignancies, including acute myeloid leukemia (AML). We used a co‐culture assay of endothelial and vascular smooth muscle cells (vSMC) to investigate the effects of AML‐conditioned medium on capillary networks. We investigated primary AML cells derived from 44 unselected patients and observed that for a large subset of patients, the constitutive cytokine release by the leukemic cells stimulated endothelial cell organization into capillary‐like networks, while there were only minor or no effects for other patients. We analyzed the constitutive AML cell release of 31 cytokines for all the patients and performed a hierarchical cluster analysis of the cytokine profile which identified two major patient subsets that differed in their ability to enhance capillary‐like networks; increased capillary‐like networks was then associated with high constitutive release of several cytokines and especially high levels of several pro‐angiogenic chemokines. Significantly increased network formation was not seen for any of the 11 acute lymphoblastic leukemia patients investigated. The cytokine response by activated normal T cells inhibited endothelial network formation in our in vitro model of angiogenesis and activated normal monocytes had only a minor influence on tube formation. Our study shows that AML‐derived cytokines can induce the organization of endothelial cells into vessel‐like structures.