电磁水对血白细胞、红细胞、血小板及血管内皮细胞影响的研究

血管中白细胞等的粘附、聚集问题能够影响微循环的血流速度,是损伤血管内皮细胞乃至形成血栓的主要因素之一.在内毒素注射大白鼠的随机、对照实验中,发现电磁水能够减轻内毒素所致的炎症刺激,并能够降低白、红细胞和血小板的粘附、聚集,能降低白细胞的渗出,能提高红细胞的电泳率,能抑制血流速度的减慢和能够减轻血管内皮细胞的损伤.t检验,差异显著（P〈0.01）以及差异明显（P〈0.05）.揭示电磁水能够提高血细胞和血管内皮细胞的表面负电荷密度,并可以减轻外因（如,内毒素）对体内细胞和血管的损伤.说明电磁水能够改善微循环,维系正常血流和防止血栓形成.     

内毒素血症大鼠肠系膜微循环中血管内皮细胞与白细胞的粘附性变化

本实验采用中文吖啶橙荧光标记技术,结合微循环观察用显微超高速摄录像装置,观察了内毒素对微血管内白细胞与微静脉血管内皮细胞的粘附性的影响。结果表明,内毒素对大鼠的血压、微血管口径和微动脉血流速度影响不大,微静脉血流速度在滴注内毒素后４５和６０ｍｉｎ下降了１６．６７％和１７．９５％（Ｐ＜０．０５）;但内毒素能迅速改变微静脉内的白细胞流态,明显增加附壁滚动的白细胞数和粘附白细胞密度指数,经测量同一微静脉内的白细胞和红细胞流速,求得白细胞与微静脉内皮细胞之间的破裂力在５ｍｉｎ和１５ｍｉｎ时下降了２５．９６％和４２．８８％（Ｐ＜０．０１）,下降趋势持续整个实验过程;说明内毒素能明显地增加白细胞与微静脉血管内皮细胞之间的粘附力。由此提示,研究白细胞与微静脉血管内皮细胞之间粘附力增强机制及寻找其抑制因素对改善微循环紊乱、抢救休克具有重要的临床意义。     

肝素钠对肢体缺血/再灌注损伤大鼠肠系膜微循环的保护作用

目的:通过观察肝素钠对肢体缺血/再灌注(LI/R)过程中肠系膜微循环的动态变化和血液流变性的影响,探讨肝素减轻LI/R损伤的可能机制,为LI/R损伤的防治提供理论依据。方法:实验采用本室常规方法复制大鼠LI/R模型,正常雄性Wistar大鼠20只,随机分为2组(n=10):肝素组(H组)和单纯缺血/再灌注组(I/R组),两组动物均于再灌注损伤后2h时动态观察肠系膜微血管管径、血流速度、白细胞黏附、白微栓及微血管壁的完整性(管周出血)等情况,同时测定各组动物血液流变学指标和血清中P-选择素和细胞间粘附分子1(ICAM-1)的值。结果:肠系膜微动、静脉管径扩张,血流速度减慢,微血管中大量白细胞贴壁、粘附,白微栓形成增多,与I/R组比较,H组大鼠肠系膜微动脉血流速度(AFV)和微静脉血流速度(VFV)显著下降(P0.01);血浆黏度(ηp)、全血低切还原黏度(Lηre)、全血高切还原黏度(Hηre)、红细胞压积(Hct)、红细胞聚集指数(EAI)、血沉方程K值(ESRK)、红细胞刚性指数(TK)均显著下降(P0.01);红细胞变形指数(EDI)显著升高(P0.01);血清中P-选择素、ICAM-1水平均显著下降(P0.01)。结论:肝素可能通过降低血清中P-选择素和ICAM-1的水平而改善肢体缺血/再灌注损伤大鼠的全身微循环状态。     

降纤酶与抗栓酶-3号联合应用治疗脑梗死40例疗效观察

降纤酶是国产蛇毒类新型溶栓抗凝药物 ,它是采用高科技分离提纯的单一组份酶制剂 ,能降解血浆纤维蛋白原 (FG) ,减少血小板粘附聚集 ,促进血管内皮细胞释放 t- PA (组织纤溶酶原激活物 ) ,降低 PAL - I (组织纤溶酶原激活物 - l) ,使形成的纤维蛋白很快被清除 ,并能抑制红细胞聚集 ,缩短红细胞通过时间 ,从而起到降低全血粘度 ,改善微循环 ,加速血栓溶解 ,使阻塞的血管再通和防止血管再栓塞的作用。而抗栓酶 - 3号已被广泛应用于临床多年 ,具有抗凝、溶栓、去纤、抗血小板粘附、聚集等作用。我院于 1 998～ 1 999年联合应用降纤酶与抗栓…     

几种不同刺激对血管内白细胞粘附的影响

目的：研究几种刺激引起白细胞与内皮细胞粘附的变化。方法：本实验采用脉冲电刺激、缺血再灌、内毒素和白介素-8等物理或药物的作用,观察大鼠肠系膜细静脉内白细胞粘附及白细胞和血管内皮粘附之间的差别。结果：缺血再灌、内毒素、内毒素、脉冲电刺激和白介纱-8（IL-8）作用后肠系膜细静脉白细胞粘附数量比正常组明显增多,IL-8用药后30min细静脉内白细胞粘附数量最多、缺血再灌、内毒素、脉冲电刺激后白细胞粘附数量大致相同。结论：缺血再灌、内毒素、脉冲电刺激能诱导白细胞的粘附作用。造成内皮损伤,IL-8诱导白细胞的粘附作用最强。     

体外流动剪切力作用下的白细胞-内皮细胞动态粘附

建立一个用于体外研究特定流动剪切力作用下白细胞和内皮细胞动态相互作用的方法。利用建立的平板流动小室系统可在体外产生特定的流动剪切力。将培养的人脐静脉内皮细胞装入平板流动小室后 ,以 0 .71dynes/cm2 的流动剪切力把含有吖啶橙染色的白细胞的灌流液导入流动小室 ,由此产生了白细胞和内皮细胞的动态粘附过程。整个粘附过程通过OlympusIX70倒置荧光显微系统观察 ,同时通过CCD摄象头录像。然后用图象采集卡将录像采集为数字图象并保存。利用针对实验设计的图象处理和分析方法 ,对采集的数字图象进行处理和测量 ,可以得到粘附白细胞的个数和滚动白细胞的速度。通过研究内毒素脂多糖 (LPS)对内皮细胞粘附功能的促进及地塞米松 (DXM )对该刺激的抑制作用来验证。对于用内毒素脂多糖 (LPS)处理的内皮细胞 ,固定粘附和慢速滚动的白细胞的个数比对照组分别显著增加了 2 3.7倍和 4 .1倍 ,同时白细胞在粘附作用过程中慢速滚动和快速滚动的速度比对照组明显降低了 2 5 .6 %和 2 6 .1%。而对于脂多糖和地塞米松 (DXM)处理过的内皮细胞 ,上述内毒素引起的影响被显著抑制了。该方法可以用于研究不同的化学和物理刺激对内皮细胞功能的影响机制 ,及用来评价各类抑制内皮细胞粘附功能的药物。     

微生态制剂益康口服液对微循环和血栓形成的影响

目的：观察益康口服液(双歧杆菌、乳酸链球菌、人参、茯苓、黄芪等)对微动脉(A)、微静脉(V)及微静脉血流速度(v)的作用和大鼠血栓形成的影响。方法：体微循环实验观察微生态制剂益康口服液对正常小鼠耳廓微循环的影响;利用大鼠体外颈总动脉-颈外静脉血流旁路法形成血小板血栓,观察益康口服液对大鼠血栓形成的影响。结果：益康口服液能明显扩张微动脉、微静脉,明显加快微静脉血流速度;提示其具有较好的改善微循环作用。结论：益康口服液能够抑制大鼠血栓的形成,提示其具有抗血小板粘附聚集作用。     

肝郁脾虚证模型大鼠血流变及TXB2、PGFla的变化

目的:探查中医肝郁脾虚证模型的血流变及相关调节因子的状态。方法:采用慢性束缚应激+过度疲劳+饮食失节法建立大鼠肝郁脾虚证模型,测定大鼠造模三周、自然恢复一周时的血流变和血浆TXB2、PGF1a。结果:与正常组相比,模型组大鼠造模三周150/s、38/s、10/s、5/s切变率下的全血粘度、还原粘度均显著升高(P<0.001),红细胞聚集指数显著降低(P<0.001),红细胞压积显著升高(P<0.01),红细胞变形指数无显著性差异(P>0.05);血浆TXB2显著升高(P<0.001),6-keto-PGF1a显著降低(P<0.05),TXB2/PGF1a显著升高(P<0.01);模型组大鼠第四周150/s、38/s、10/s、5/s切变率下的全血粘度、还原粘度仍显著升高(P<0.001或P<0.01);红细胞聚集指数显著降低(P<0.001);红细胞压积与变形指数无显著性差异(P>0.05);血浆TXB2和TXB2/PGF1a显著降低(P<0.05),6-keto-PGF1a显著升高(P<0.05)。结论:肝郁脾虚证大鼠存在血液高粘和血栓易形成状态,恢复期血液高粘同时伴有扩血管因素的加强。提示肝郁脾虚证有血流...     

血管壁剪切应力系统及用于内皮细胞与白细胞粘附的研究

本文建立了对血管壁上的内皮细胞施加剪切应力的系统。通过对系统中的血管内流场分析表明,该系统中的血管段的中间部分作为研究剪切应力对内皮细胞作用,以及研究内皮细胞与其它细胞粘附力的场所是比较理想的。采用该系统对在体内皮细胞研究发现：当对内皮细胞施加２８ｄｙｎ／ｃｍ２的剪切应力时,内皮细胞并未出现暴发性释放前列环素的现象,前列环素释放水平略有升高（０．４１±０．０５ｎｇ／ｃｍ２ｍｉｎ）以后呈线性下降并稳定在一定水平上（０．１７±０．０４ｎｇ／ｃｍ２ｍｉｎ）。在２８ｄｙｎ／ｃｍ２剪切应力下,受机械损伤的内皮细胞和动脉粥样硬化的内皮细胞仍能与较多白细胞粘附,而在正常内皮细胞和去掉内皮细胞的动脉壁几乎无白细胞粘附,说明在内皮细胞受损或动脉粥样硬化时,内皮细胞与白细胞的粘附增强     

蝮蛇毒蛋白C激活物对内毒素性离体大鼠心脏功能的影响

目的研究蝮蛇毒蛋白C激活物(PCA)组分对大鼠内毒素(LPS)性心肌损伤作用的影响。方法取SD雄性大鼠32只随机分成正常对照组、LPS组、PCA组和PCA LPS组,用Krebs-Henseleit(K-H)液对大鼠离体心脏行主动脉逆灌。在相应时点记录HR、LVSP、LVEDP、LVDP、 dp/dtmax、-dp/dtmax的变化,并测定冠脉流出液中的超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量。结果PCA明显减轻LPS诱导的心功能改变并抑制心肌SOD活性降低和MDA的升高(P<0.01)。结论PCA对内毒素诱导的心肌损伤改善作用明显,其机制可能是通过保护血管内皮功能,改善微循环,稳定心肌酶活性和膜相结构等途径有关。     

Alterations by leukocytes of erythrocyte flow in microchannels

The effects of leukocytes on blood rheology in the microcirculation may be predicted to result from the rheological characteristics of the individual white blood cell types and their behavior at entrances of small vessels; their influence on flow resistance of blood due to their disturbance of erythrocyte flow; and the obstruction caused by their adherence to endothelial cells or geometrical hindrance of their flow at irregular or narrow regions. Deformation of leukocytes in micropipettes indicates the relative rigidity of lymphocytes and blast cells from leukemias in contrast to granulocyte viscoelastic properties, and entry times for lymphocytes in 2.6-4.6-microns channels were 11-151 s but milliseconds in 9.1-micron tubes. Lymphocytes and erythrocytes rarely (less than 1%) adhered to cultured endothelium; however, granulocytes adhered avidly at shear stresses of 10-100 dyn/cm2, typical of microcirculation. In the 9.1-microns flow tube at Hct = 17.7 +/- 2.6%, increasing [WBC] caused a linear increase in flow resistance, but above [WBC] = 1000/mm3 resistance tended to plateau. These data support the interpretation that granulocyte adherence to endothelial cells with the potential for obstruction of flow may be a more significant rheological consequence of leukocytes in blood than their resistance to flow because of their relative rigidity or their influence on flow resistance by perturbation of the erythrocyte flow.     

Postischaemic circulation disturbances.

Restoration of blood supply after ischaemic conditions in extremities and testes is inhibited by reversible intravasal aggregation of erythrocytes. This process is promoted by the increased permeability of the capillaries associated with the formation of oedema and the entailing increase of the haematocrit. For overcoming the stasis the increased structural viscosity caused by the aggregation of erythrocytes requires an increase in pressure as a starter effect which is not achieved by the flow pressure at once everywhere. Intravenously administered particles of Indian ink mark the formation and dissolution of aggregates. Even areas with originally normal blood supply may be obstructed by the later formation of aggregates. Thrombi on the walls of arterial and venous vessels and other lesions of the intima do not sufficiently explain the disturbance of perfusion. Oedema and extravasating leucocytes are found in the microcirculation. The parenchyma to be supplied shows formation of necrosis.     

A comparison of the effects of photodynamic therapy on normal and tumor blood vessels in the rat microcirculation

The effects of light activation of the tumor photosensitizer dihematoporphyrin ether (DHE) were studied in the microcirculation of the rat cremaster muscle. Arterioles and venules in an implanted chondrosarcoma were studied by in vivo television microscopy and were compared to normal vessels of the same size elsewhere in the preparation and in control preparations. Activation with green light (530-560 nm, 200 mW/cm2, 120 J/cm2) 48 h after intraperitoneal injection of DHE (10 mg/kg body wt) resulted in significant narrowing of diameters of red blood cell columns in tumor arterioles and venules. The response in normal and control arterioles and venules was not significantly different from that seen in the tumor vessels except that the control arterioles did not remain significantly constricted during the treatment period. Treatment resulted in stasis of blood flow in 90% of tumor and normal arterioles at the completion of light activation. In venules, stasis of blood flow was observed in 75% of tumor and 70% of normal vessels. Vasoconstriction was the primary response in arterioles, while thrombosis predominated in venules. Morphologic assessment of light-activated vessels in the cremaster preparation by transmission electron microscopy revealed platelet aggregation with damage to endothelial cells and smooth muscle cells. Perivascular effects observed included interstitial edema and damage to skeletal muscle cells. In the tumor-bearing preparation, no direct cytotoxic effect on the tumor cells was shown. The surrounding vessels exhibited similar vascular stasis, but the lining cells appeared minimally affected. Photoactivation of DHE results in significant changes in the microcirculation which lead to stasis of blood flow. In this model, the response was similar for the normal microvasculature and for the microcirculation of an implanted chondrosarcoma. These effects may account, in part, for the mechanism of action of photodynamic therapy.     

Nitric oxide influences red blood cell velocity independently of changes in the vascular tone

Abstract

Nitric oxide (NO) plays a key role in regulation of vascular tone and blood flow. In the microcirculation blood flow is strongly dependent on red blood cells (RBC) deformability. In vitro NO increases RBC deformability. This study hypothesized that NO increases RBC velocity in vivo not only by regulating vascular tone, but also by modifying RBC deformability. The effects of NO on RBC velocity were analysed by intra-vital microscopy in the microcirculation of the chorioallantoic membrane (CAM) of the avian embryo at day 7 post-fertilization, when all vessels lack smooth muscle cells and vascular tone is not affected by NO. It was found that inhibition of enzymatic NO synthesis and NO scavenging decreased intracellular NO levels and avian RBC deformability in vitro. Injection of a NO synthase-inhibitor or a NO scavenger into the microcirculation of the CAM decreased capillary RBC velocity and deformation, while the diameter of the vessels remained constant. The results indicate that scavenging of NO and inhibition of NO synthesis decrease RBC velocity not only by regulating vascular tone but also by decreasing RBC deformability.     

Nitric oxide influences red blood cell velocity independently of changes in the vascular tone

Nitric oxide (NO) plays a key role in regulation of vascular tone and blood flow. In the microcirculation blood flow is strongly dependent on red blood cells (RBC) deformability. In vitro NO increases RBC deformability. This study hypothesized that NO increases RBC velocity in vivo not only by regulating vascular tone, but also by modifying RBC deformability. The effects of NO on RBC velocity were analysed by intra-vital microscopy in the microcirculation of the chorioallantoic membrane (CAM) of the avian embryo at day 7 post-fertilization, when all vessels lack smooth muscle cells and vascular tone is not affected by NO. It was found that inhibition of enzymatic NO synthesis and NO scavenging decreased intracellular NO levels and avian RBC deformability in vitro. Injection of a NO synthase-inhibitor or a NO scavenger into the microcirculation of the CAM decreased capillary RBC velocity and deformation, while the diameter of the vessels remained constant. The results indicate that scavenging of NO and inhibition of NO synthesis decrease RBC velocity not only by regulating vascular tone but also by decreasing RBC deformability.     

Monoclonal antibody OKM5 inhibits the in vitro binding of Plasmodium falciparum-infected erythrocytes to monocytes, endothelial, and C32 melanoma cells

Plasmodium falciparum-infected erythrocytes bind in vitro to human endothelial cells, monocytes, and a certain melanoma cell line. Evidence suggests that this interaction is mediated by similar mechanisms which lead to the sequestration of parasitized erythrocytes in vivo through their attachment to endothelial cells of small blood vessels. We show here that monoclonal antibody OKM5, previously shown to react with the membranes of endothelial cells, monocytes, and platelets, also reacts with the C32 melanoma cell line which also binds P. falciparum-infected erythrocytes. At relatively low concentrations, OKM5 inhibits and reverses the in vitro adherence of infected erythrocytes to target cells. As with monocytes, OKM5 antibody recognizes an 125I-labeled protein of approximately 88 Kd on the surface of C32 melanoma cells. It seems likely, therefore, that the 88 Kd polypeptide plays a role in cytoadherence, possibly as the receptor or part of a receptor for a ligand on the surface of infected erythrocytes.     

滇丹参、甘西鼠尾、褐毛甘西鼠尾水提取液对鼠血栓形成及微循环的影响

目的：观察云南产甘西鼠尾属药物滇丹参、甘西鼠尾、褐毛甘西鼠尾的水提取物对鼠血栓形成及微循环的影响。方法：用电刺激法造成大鼠颈动脉血栓形成,观察血流阻塞时间(OT值);小鼠尾静脉注射血栓形成诱导剂,观察滇丹参、甘西鼠尾、褐毛甘西鼠尾对抗肺血栓形成的作用;采用活体微循环观察方法,研究滇丹参、甘西鼠尾、褐毛甘西鼠尾对去甲肾上腺素和高分子右旋糖苷所致小鼠、大鼠肠系膜微循环障碍的影响。结果：滇丹参、甘西鼠尾、褐毛甘西鼠尾2、4g/kg时,能明显延长闭塞性血栓形成时间(OT值)(P＜0．01)。20、40g/kg时,能有效抑制胶原和肾上腺素复合液所致小鼠肺血栓形成(P＜0．0l,P＜0．05)。能显著改善微动脉、微静脉痉挛和血液流态,增加毛细血管开放数,加快血流速度(P＜0．0l,P＜0．05)。结论：滇丹参、甘西鼠尾、褐毛甘西鼠尾均能防止血栓形成、改善微循环障碍。     

电针改善VBI所致内耳微循环障碍及对VOR的影响

目的：对照电针和西比灵（盐酸氟桂利嗪）的效应,探讨电针改善椎-基底动脉供血不足（VBI）所致内耳微循环障碍的机理及电针对前庭-眼反射（VOR）的影响。方法：组织硬化剂-775注射液注射致家兔左侧颈椎横突软组织建立椎动脉型颈椎病（VCS）模型,采用直线加速度（LA）和水平旋转（HR）诱发眼震电图（ENG）、经颅多谱勒（TCD）、激光多谱勒血流计（LDF）和血液流变学,检测VBI家兔NEG频率、基底动脉（BA）血流速度、内耳血流量（IEBF）、血液粘度和细胞流变性变化。结果：模型组ENG频率、BA血流速度和IEBF显著降低,全血中、低切变率粘度和红细胞聚集指数（EDI）显著增高。西比灵可显著降低全血粘度和EDI,加快BA收缩期血流速度（Vs）,但对舒张期血流速度（Vd）和平均血流速度（Vm）无明显作用。电针不能降低血液粘度和EDI无明显作用,但改善IEBF和LA诱发的ENG作用较西比灵显著,并有增快Vs、Vd和Vm的趋势,电针和西比灵对HR诱发的ENG均无改善作用。结论：VBI所致内耳微循环障碍可引起前庭和水平半规管功能障碍,电针可能通过神经一体液调节改善VBI,并可能主要通过增强内耳微循环局部调节改善内耳血供,恢复前庭囊斑对LA的敏感性。电针效应优于西比灵。VBI模型前庭器官内血流分配及前庭信号转导可能存在复杂的机制,有待深入研究。     

Influence of gravity on flow distribution of red blood cells in microcirculation

The influence of the gravity on flow distribution of erythrocytes in microcirculation was examined. We developed a new centrifuge system with a rotation disc. An observation system of blood flow in a micro-flow channel was arranged on the disc. Erythrocyte flow in the micro-flow tube was displaced under the gravity. This study suggests that the gravity affects the transfer of substances from blood vessels to tissues.     

Microcirculation-on-a-Chip: A Microfluidic Platform for Assaying Blood- and Lymphatic-Vessel Permeability

We developed a microfluidic model of microcirculation containing both blood and lymphatic vessels for examining vascular permeability. The designed microfluidic device harbors upper and lower channels that are partly aligned and are separated by a porous membrane, and on this membrane, blood vascular endothelial cells (BECs) and lymphatic endothelial cells (LECs) were cocultured back-to-back. At cell-cell junctions of both BECs and LECs, claudin-5 and VE-cadherin were detected. The permeability coefficient measured here was lower than the value reported for isolated mammalian venules. Moreover, our results showed that the flow culture established in the device promoted the formation of endothelial cell-cell junctions, and that treatment with histamine, an inflammation-promoting substance, induced changes in the localization of tight and adherens junction-associated proteins and an increase in vascular permeability in the microdevice. These findings indicated that both BECs and LECs appeared to retain their functions in the microfluidic coculture platform. Using this microcirculation device, the vascular damage induced by habu snake venom was successfully assayed, and the assay time was reduced from 24 h to 30 min. This is the first report of a microcirculation model in which BECs and LECs were cocultured. Because the micromodel includes lymphatic vessels in addition to blood vessels, the model can be used to evaluate both vascular permeability and lymphatic return rate.