血管内皮细胞缺氧性损伤及药物干预作用的实验观察

缺血缺氧均能使血管内皮细胞受损。本研究采用新生儿脐静脉血管内皮细胞体外原代培养方法，并施加缺氧条件，观察细胞培养上清液和细胞匀浆中丙二醛（MDA）含量变化及中药川芎嗪对它的影响。结果显示：血管内皮细胞在缺氧环境中，上清液和细胞匀浆中MDA含量明显增加（P＜0.001），加入不同浓度川芎嗪后均可阻止上述现象的发生。实验结果表明川芎嗪可明显抑制自由基的产生及细胞的脂质过氧化作用，对血管内皮细胞抗缺氧损伤具有保护作用。     

三七皂苷对氧化低密度脂蛋白损伤血管内皮细胞保护作用的研究

目的:研究三七皂苷(PNS)对氧化低密度脂蛋白(Ox-LDL)损伤血管内皮细胞的保护作用。方法:体外培养人脐静脉内皮细胞(HUVEC),加入100mg·L-1Ox-LDL造成细胞损伤,同时加入不同浓度PNS进行干预。观察形态学变化;采用MTT法测定细胞活性;比色法检测细胞上清液中乳酸脱氢酶(LDH)的释放量;硝酸还原法和放射免疫法分别测定HUVEC培养上清液中一氧化氮(NO)、内皮素(ET)含量。结果:各剂量PNS可使作用Ox-LDL的HUVEC形态趋于正常。与正常对照组比较,HU-VEC经Ox-LDL作用后,细胞活性显著下降,LDH的释放量显著升高;300、100、30mg·L-1PNS显著升高细胞活性,抑制LDH释放。HUVEC经Ox-LDL作用后,与正常对照组比较,培养上清液中NO含量显著降低,ET含量显著升高;300、100mg·L-1PNS显著升高NO含量,300mg·L-1PNS显著降低ET含量。结论:PNS对Ox-LDL损伤的血管内皮细胞具有一定的保护作用,可调节内皮细胞活性物质NO、ET的分泌。     

川芎嗪对缺氧时视网膜血管内皮细胞HIF-1α、VEGF表达及细胞增殖的影响

目的研究川芎嗪对缺氧时体外培养的牛眼视网膜血管内皮细胞（BREC）VEGF表达及细胞增殖活性的影响,探讨川芎嗪治疗视网膜新生血管性疾病的药理机制及可行性。方法 CoCl2模拟缺氧处理培养的BREC细胞,用浓度分别为20、40、120μg/mL的川芎嗪分别加入细胞培养液中24h,采用ELISA法检测细胞培养上清液VEGF含量,免疫组织化学法和生物荧光法分别检测细胞PCNA的表达、细胞内ATP含量以分析细胞增殖能力,采用免疫组织化学法检测HIF-1α的表达。结果川芎嗪可减少缺氧引起的视网膜血管内皮细胞中PCNA的表达,降低细胞内ATP含量,降低培养上清液中的VEGF浓度,且呈剂量依赖性。结论川芎嗪可抑制缺氧引起的视网膜血管内皮细胞的增殖。其机制可能是通过抑制HIF-1α途径来实现的。     

前列地尔对兔内皮细胞缺氧复氧损伤后抗栓功能的影响

目的：探讨前列地尔（PGE1）对兔内皮细胞缺氧复氧损伤模型的抗栓功能影响。方法对体外培养、并建立兔内皮细胞缺氧复氧损伤的模型进行分组：模型组、低浓度PGE1组、中浓度PGE1组、高浓度PGE1组、正常组。对以上各组培养上清液中一氧化氮（NO）、超氧化物歧化酶（SOD）、丙二醛（MDA）、细胞内血管假性血友病因子（vWF）的含量或表达等进行测量。结果与正常组比较，模型组NO含量、SOD活性、vWF表达均有所下降， MDA含量增加，且差异具有统计学意义（P〈0.05）；而低、中、高浓度PGE1组与模型组比较，其NO含量、SOD活性、vWF表达均有所增加或增强， MDA含量降低，且差异具有统计学意义（P〈0.05）。结论 PGE1对机体缺氧复氧损伤之后的血管内皮细胞抗栓功能具有积极的保护作用。     

川芎嗪和尼莫地平对血管内皮细胞分泌ET和NO的影响

为探讨川芎嗪和尼莫地平在心脑血管疾病防治中的作用机制,我们采用内皮细胞体外培养的方法,观察尼莫地平和川芎嗪对内皮细胞分泌内皮素（ET）和一氧化氮（NO）的影响。实验结果发现,两药均能不同程度地抑制正常体外培养的内皮细胞产生内皮素,改变ET和NO的比例,从而起到防治心脑血管疾病的作用。     

金钠多与艾司洛尔对缺氧联合Ang-2所致血管内皮细胞分泌内皮素的保护作用

目的 比较金钠多与艾司洛尔对缺氧联合血管紧张素2(Ang-2)所致血管内皮细胞分泌内皮素(ET)的保护作用.方法 将培养的血管内皮细胞分为四组:A组缺氧前加入10-5mol/L的Ang-2;B组在A组基础上加入10mg/L的金钠多;C组在A组基础上加入5 mg/L的艾司洛尔;D组正常培养作对照.用放射免疫法测定作用前、作用后0.5、6、24h各组ET含量.结果 与D组比较,缺氧后0.5、6、24 h三个时间点A组的ET含量均增高(P<0.01),B、C两组ET比A组降低(P<0.01).用药后0.5 h,B组ET比C组降低(P<0.05),用药后6 h和24 h更明显(P<0.01).结论 金钠多和艾司洛尔对缺氧联合Ang-2对血管内皮细胞的损害均有保护作用,金钠多的保护作用更持久.     

Omapatrilat对AngⅡ致人血管内皮细胞损伤的保护作用研究

目的观察Omapatrilat对血管紧张素Ⅱ(AngⅡ)致人脐静脉内皮细胞(HUVECs)损伤的保护作用。方法体外培养HUVECs,随机分为4组:空白对照组,AngⅡ组,Omapatrilat组和AngⅡ+Omapatrilat组。分光光度计测定培养的HUVECs乳酸脱氢酶(LDH)的漏出,流式细胞仪技术检测细胞周期,硝酸还原酶法和放射免疫分析技术分别测定HUVECs上清液中一氧化氮(NO)和内皮素(ET1)含量。结果10-7mol·L-1AngⅡ可增加HUVECsLDH漏出和ET1释放,Omapatrilat(10-6mol·L-1)对此具有明显抑制作用,同时它可促进HUVECs增殖和NO释放。结论Omapatrilat可减弱AngⅡ导致的体外培养HUVECs损伤,对内皮细胞具有保护作用。     

淫羊藿总黄酮对LPC诱导内皮细胞损伤的保护作用

目的观察淫羊藿总黄酮对溶血性磷脂酰胆碱（LPC）诱导血管内皮细胞损伤的保护作用。方法将血管内皮细胞分为正常对照组,LPC损伤对照组,LPC损伤加淫羊藿总黄酮低、中、高浓度（50、100、200nag·L^-1）组,检测细胞上清液中一氧化氮（NO）、丙二醛（MDA）、乳酸脱氢酶（LDH）的含量和细胞内活性氧（ROS）水平。结果淫羊藿总黄酮剂量依赖性地降低LPC诱导的内皮细胞上清液中MDA、LDH含量增加并抑制细胞内ROS活性,且剂量依赖性地增加细胞上清液中NO含量。结论淫羊藿总黄酮对LPC诱导的血管内皮细胞损伤具有保护作用,其机制与抑制细胞内ROS活性有关。     

川芎嗪对高血压患者血管内皮细胞功能的影响

目的探讨川芎嗪对高血压患者血管内皮细胞功能的影响。方法选择佛山市南海区第四人民医院2009年2月至2011年3月收治的高血压患者50例作为治疗组,给予川芎嗪120mg静脉滴注,1次/d,15d为1个疗程;另选同期于佛山市南海区第四人民医院进行健康体检者50例作为对照组。检测两组的血浆内皮素-1(ET-1)、一氧化氮(NO)含量,比较治疗组治疗前后的ET-1和NO含量。结果与对照组比较,治疗组治疗前的ET-1含量明显明显高于对照组(P<0.05),NO含量明显低于对照组(P<0.05);治疗后,治疗组的ET-1含量明显降低,NO含量明显升高,与治疗前比较差异具有统计学意义(P<0.05)。结论高血压患者血管内皮细胞功能紊乱,血浆ET-1/NO比例失调。川芎嗪具有保护血管内皮细胞功能,值得在临床治疗高血压中推广应用。     

氯沙坦对血管紧张素Ⅱ致培养的牛脑微血管内皮细胞损伤的保护作用

目的：观察氯沙坦对血管紧张素Ⅱ（AngⅡ）致牛脑微血管内皮细胞（BCMECs）损伤的保护作用。方法：用分光光度计测定培养的BCMECs乳酸脱氢酶（LDH）的漏出量，流式细胞仪测定BCMECs细胞间粘附分子－1（ICAM－1）的表达量，硝酸还原酶法和放射免疫分析法分别测定BCMECs上清液中一氧化氮（NO）和内皮素－1（ET1）的含量。结果：AngⅡ呈剂量依赖性增加BCMECsLDH漏出、NO和ET1释放及ICAM－1表达，氯沙坦对此均有明显抑制作用。结论：氯沙坦抑制 AngⅡ致体外培养BCMECs的损伤。     

不同浓度乙醇对内皮细胞分泌作用的影响

目的 研究不同浓度乙醇对内皮细胞的保护作用。方法 应用过氧化氢损伤培养的人脐静脉内皮细胞,并与不同浓度乙醇进行孵育,检测内皮细胞产生的内皮素及一氧化氮。结果低、中浓度乙醇明显减少损伤的内皮细胞产生的内皮素。同时实验还提示长时间高浓度乙醇对内皮细胞有明显损伤作用;不同浓度的乙醇均抑制内皮细胞产生一氧化氮。结论低、中浓度乙醇使内皮细胞在受损时生成的内皮素明显减少。     

妊高征患者血一氧化氮、内皮素和肿瘤坏死因子检测的临床意义

目的　探讨一氧化氮 (NO)、内皮素 (ET)和肿瘤坏死因子 (TNF)含量在妊高征中含量的变化。方法　采用放射免疫分析法检测了 38例妊高征患者血清中NO、ET和TNF含量并与 35名正常孕妇作比较。结果　妊高征患者血中NO水平降低 (P <0 .0 1) ,ET、TNF水平则显著地升高 (P <0 .0 1) ,且与病情的轻重密切相关。结论　妊高征患者血管内皮细胞存在内分泌功能紊乱 ,ET合成增加 ,NO释放减少 ,TNF分泌过多 ,三者反馈调控失衡在妊高征的发病中具有重要的临床价值     

妊高症患者内皮素、一氧化氮、肿瘤坏死因子联合测定的临床意义

目的：探讨内皮素、一氧化氮,肿瘤坏死因子在妊高症发病中的作用。方法：放免法测定了32例妊高症患血中ET,TNF含量,同时采用Greiss法测定亚硝酸盐及硝酸盐以间接测定NO含量,并与30名正常晚孕妇女作比较。结果：妊高症患产前ET,TNF含量较正常妊娠组明显升高（P＜0．01）,NO则明显下降（P＜0．01）,与妊高症的病情程度有一定的关系,产后逐渐恢复正常,结论：妊高症患血管内皮细胞存在内分泌功能紊乱,ET,TNF合成释放增加,NO释放减少,以及三反馈调控失衡在妊高症发病中具有重要意义。     

Endothelin receptor-mediated vasodilatation: effects of organ culture

Culture of intact arteries is a frequently employed experimental model for investigating the mechanisms governing the regulation of vascular endothelin receptors. Endothelin type A (ET(A)) and type B (ET(B)) receptors on vascular smooth muscle cells are up-regulated in organ culture and the enhanced vasoconstriction mimics the changes that occur in cardiovascular disease. The effect of organ culture on endothelial dilatory endothelin ET(B) receptors is not known. We hypothesize that organ culture decreases the endothelin receptor-mediated dilatation and that this is one possible mechanism by which the effects of the endothelin in blood vessels are altered during culture. Porcine coronary arteries were studied before and after 24 h of culture, using in vitro pharmacology and immunofluorescence. Sarafotoxin 6c and endothelin-1 were used to examine the endothelin ET(A) and ET(B) receptor effects, and the antagonists, Nomega-nitro-l-arginine (l-NOARG) for nitric oxide (NO), indomethacin for prostaglandins and charybdotoxin in combination with apamin for endothelium-derived hyperpolarizing factor (EDHF), were used to study the endothelium-derived dilatory mediators. Organ culture induced up-regulation of the sarafotoxin 6c (ET(B) receptor agonist) and endothelin-1 (ET(A) receptor agonist) elicited vasoconstriction. The sarafotoxin 6c contraction was stronger after endothelium denudation, suggesting endothelium-dependent dilatation. The endothelin-1 contraction was not affected by endothelium denudation. The increase in sarafotoxin 6c contraction after removal of the endothelium was more pronounced before than after organ culture, suggesting down-regulated endothelial endothelin ET(B) receptors. Also, the immunofluorescence staining intensities for endothelial endothelin ET(B) receptors were higher before than after organ culture. Pre-incubation with inhibitors for dilatory mediators suggested that both NO and EDHF play a vasodilatory role, while prostaglandins are not involved. In conclusion, endothelial endothelin ET(B) receptors induce NO and EDHF mediated vasodilatation in porcine coronary arteries. In organ culture, endothelial endothelin ET(B) receptors are down-regulated, mimicking the changes that occur in cardiovascular disease. Down-regulation of endothelial endothelin ET(B) receptors may in part explain the increased endothelin ET(B) receptor-mediated vasoconstriction frequently studied in organ culture.     

支气管哮喘患儿血内皮素、一氧化氮含量测定的临床意义

目的　探讨了支气管哮喘患儿血内皮素和一氧化氮含量的变化。方法　应用放免法和化学法测定了 38例支气管哮喘患儿血内皮素和一氧化氮含量、并与 35名正常健康人作比较。结果　支气管哮喘患儿血内皮素水平明显升高 (P<0 .0 1)。一氧化氮水平明显下降 (P<0 .0 1)且与患儿病情程度有关。结论　支气管哮喘患儿血管内皮细胞存在内分泌功能的紊乱 ,ET合成释放增加 ,一氧化氮释放减少 ,以及两者反馈调控失衡在支气管哮喘的发病中具有重要意义     

Endothelin-1 enhances vascular cell adhesion molecule-1 expression in tumor necrosis factor alpha-stimulated vascular endothelial cells

Vascular cell adhesion molecule-1 (VCAM-1) is a mononuclear leukocyte-selective adhesion molecule that is expressed in human vascular endothelial cells at sites of local inflammation. It participates in local endothelial-monocyte interactions during the initiation of atherosclerosis. In the present study, endothelin alone did not induce the surface expression and mRNA accumulation of VCAM-1 in human vascular endothelial cells, but inhibition of endogenous nitric oxide (NO) by N(G)-monomethyl-L-arginine enhanced the surface expression and mRNA accumulation of VCAM-1 stimulated by endothelin-1. It is conceivable that in human vascular endothelial cells, stimulation of an endothelin receptor results in the production of nitric oxide (NO), suppressing the expression of VCAM-1. Endothelin-1 enhanced the surface expression and mRNA accumulation of VCAM-1 in cells treated with tumor necrosis factor alpha (TNF-alpha). The enhancement by endothelin-1 may be explained by the inhibitory effect of TNF-alpha on endothelin-induced NO production. Pretreatment with BQ788 (an endothelin ET(B) receptor antagonist) or inhibitors of nuclear factor kappa B (NF-kappaB) activation completely diminished the synergistic enhancement of VCAM-1 expression by endothelin-1 in TNF-alpha-stimulated vascular endothelial cells, both at the protein and mRNA levels. These findings suggest that the synergistic enhancement of VCAM-1 expression by TNF-alpha and endothelin ET(B) receptor stimulation may be augmented by the induction of NF-kappaB binding activity in human vascular endothelial cells.     

扁杏仁肽对血管内皮功能的影响

目的探究扁杏仁肽对人脐静脉内皮细胞(human umbilical vascular endothelial cells,HUVECs)内皮功能的影响。方法体外培养HUVECs,以150,300和600μg·mL~(-1)不同剂量的扁杏仁全肽、扁杏仁谷蛋白肽给药,进行细胞生长增殖试验及细胞培养液中一氧化氮(NO)和内皮素(ET)含量的检测。结果细胞培养48h后,与对照组相比,所有剂量的扁杏仁全肽和扁杏仁谷蛋白肽对HUVECs均有显著的抑制作用(P<0.01),同时显著地减弱去甲肾上腺素对细胞增殖的促进作用(P<0.01);在不同剂量扁杏仁全肽和扁杏仁谷蛋白肽干预下培养细胞48h,与对照组相比,培养液中NO含量显著升高、ET含量显著降低(P<0.01),同时对去甲肾上腺素具有显著的拮抗作用(P<0.01)。结论扁杏仁全肽特别是扁杏仁谷蛋白肽对血管内皮功能具有很强的保护作用。     

Co-operation between endothelin and nitric oxide in promoting endothelial cell migration and angiogenesis

1. Among the diverse functions of endothelins (ET), their role in the remodelling of blood vessels remains poorly examined. In the present review, we summarize findings obtained in our laboratory and present four independent lines of evidence to support this novel function. We also demonstrate that the motogenic and angiogenic effects of ET are mediated via the ETB receptor and that the functional endothelial nitric oxide synthase (NOS) is requisite for this action. 2. We demonstrated that ET stimulates transmigration of endothelial cells in a modified Boyden chamber and accelerates endothelial wound healing acting via ETB receptors. 3. In genetically engineered Chinese hamster ovary cells expressing either ETB receptor or endothelial NOS or both, application of ET results in accelerated cell migration only when the receptor and the enzyme are coexpressed. Application of antisense oligonucleotides producing a specific knockdown of the endothelial NOS results in the loss of ET ability to stimulate endothelial cell migration in response to ET. 4. Finally, using a novel model of in vivo angiogenesis, we were able to demonstrate that ET enhances formation of new vessels, but this effect requires functional endothelial NOS. 5. The described phenomenon of NO production, serving as a prerequisite for endothelial cell locomotion in response to activation of ETB receptor may explain a host of pathophysiological observations on inadequate angiogenesis despite enhanced generation of ET-1. 6. Based on the contribution of endothelial cell migration to angiogenesis, these data may implicate insufficient NO production in pathological states (e.g. atherosclerosis, heart failure and hypertension) in the inappropriate response to angiogenic stimuli.     

银杏黄酮磷脂复合物对血管内皮保护作用初探

目的:探讨银杏黄酮磷脂复合物对大鼠缺血再灌注状态下血管内皮的保护作用。方法:健康Wister大鼠40只,随机分为假手术组、模型组、银杏黄酮组及银杏黄酮磷脂复合物组,每组10只,检测各组血浆内皮素(ET1)、血清一氧化氮(NO)水平,并取心尖部心肌做电镜,观察血管内皮的显微结构变化。结果:大鼠在心肌缺血30min再灌注120min状态下,银杏黄酮磷脂复合物可明显降低血清NO含量,亦使血浆ET1水平下降,同时改善血管内皮超微结构的破坏程度,且变化较银杏黄酮组显著。结论:银杏黄酮磷脂复合物对大鼠心肌再灌注血管内皮损伤具有保护作用,可能与其减少自由基对内皮细胞的氧化损伤,减少内源性血管活性物质ET1释放有关。     

坎地沙坦西酯对高血压患者内皮功能的影响

目的:探讨坎地沙坦西酯对高血压患者内皮功能的影响。方法:选择门诊及住院的原发性高血压患者35例,在原抗高血压治疗药物基础上,加用坎地沙坦西酯8 mg qd,共8周。对照组30例,为健康正常人。两组病例于试验前和试验后8周测定血浆内皮素(ET)水平及血浆一氧化氮(NO)水平。结果:治疗组治疗后血浆ET水平明显下降,血浆NO浓度升高,与治疗前相比差异显著(P<0.001);治疗后ET及NO水平与对照组比较,差异有统计学意义(P<0.05)。治疗组用药后收缩压下降(19.43±9.01)mmHg,舒张压下降(9.36±3.12)mmHg,与治疗前比较,差异有统计学意义(P<0.05)。结论:坎地沙坦西酯在有效控制血压的同时,降低ET水平,升高NO水平,提示其具有改善内皮功能的作用。