原发性高血压大鼠血浆外泌体水平及其与微循环变化的相关性研究

目的:研究13周龄原发性高血压大鼠(SHR)外周血外泌体数量和大小分布与微循环功能的相关性。方法:市购13周龄雄性SHR(实验组)和相同周龄的WKY大鼠(对照组)各7只。采用多功能激光多普勒血流探测仪(LDF)检测两组大鼠耳廓、趾及脑皮质血流量、血细胞聚集度和血流速度,采用血流成像技术(LDPI)观察脑皮质血流分布。取外周血,提取血浆外泌体,采用透射电镜观察外泌体形态,以Apogee流式细胞术检测总外泌体水平和直径100nm的外泌体水平,分析实验组大鼠外泌体水平与其微循环指标的相关性。结果:LDF结果显示,实验组大鼠脑皮质血流量和血流速度均显著高于对照组(P0.05或P0.01),而实验组大鼠趾血流量和血流速度明显低于对照组(P0.05),两组大鼠的耳廓血流量、血流速度及耳廓、趾和脑皮质的血细胞聚集度差异均无统计学意义(P0.05);LDPI结果显示,实验组大鼠脑皮质血流灌注量显著高于对照组(P0.01)。体外实验显示,实验大鼠血浆外泌体大小分布不均,但以直径100nm者居多;实验组总外泌体水平和直径100nm外泌体水平均明显高于对照组(P0.05)。相关性分析显示,实验组大鼠血浆总外泌体和直径100nm外泌体水平与脑皮质血流量呈显著正相关(P0.01)。结论:脑微循环功能改变与血浆总外泌体含量,尤其直径100nm的外泌体水平显著相关,提示外泌体水平的升高可能预警原发性高血压发生和发展过程中的微循环变化,或可作为高血压早期的监测标记物。     

激光多普勒血流探测仪小波分析早期高血压大鼠血流灌注频谱特征

目的:利用高效激光多普勒血流探测仪(LDF)的小波分析早期自发性高血压大鼠(SHR)微循环血流灌注频谱特征。方法:8周龄、10周龄、13周龄雄性SHR大鼠(实验组)和8周龄、10周龄、13周龄WKY大鼠(对照组),每组各周龄大鼠均7只,采用多功能LDF检测两组大鼠耳、趾、脑皮质和肾皮质血流量(Flu),并利用小波分析上述部位的血流灌注信号(SFBO)的频谱特征。结果:13周龄实验组大鼠的趾部血流灌注信号在神经活动(brand4)和内皮来源(brand5)相关的信号区间内明显高于同周龄对照组大鼠(P0.05)。实验组大鼠各部位的血流灌注信号随年龄变化的趋势不明显,在趾部,10周龄实验组大鼠在各个频率信号区间的血流灌注信号均明显减弱,但是在13周时又再次升高,肾部交感神经来源(brand4)信号区间也呈现该现象。耳部和脑部的血流灌注信号随周龄的变化相对平稳。对照大鼠脑部的血流随着周龄的增加在各个频率区间都呈现血流灌注信号减弱的趋势,尤其是与内皮来源(Brand5)(P0.05或P0.01)和肌源性活动(Brand3)相关的频率范围。结论:通过小波转换分析早期高血压大鼠的激光多普勒血流灌注信号,从时频分析的角度为评估高血压大鼠的微循环状况提供另一个方向。     

全杜仲胶囊对自发性高血压大鼠肠壁微血流灌注的影响

目的 探讨全杜仲胶囊(QDZJN)和松脂醇二葡萄糖苷(PDG)对自发性高血压大鼠(SHRs)肠壁微循环血流灌注及其频谱特征的影响。方法 将SHRs随机分为SHR,SHR+QDZJN,SHR+PDG(L),SHR+PDG (M)和SHR+PDG (H)组。对照组为Wistar-kyoto(WKY)鼠。各组分别接受QDZJN或纯净水(对照组和SHR组)灌胃2个月。手术暴露大鼠小肠,激光多普勒血流探测仪(LDF)和血流成像仪(LDPI)检测并比较各组大鼠肠壁微血流灌注水平。小波分析比较各组肠壁内皮源性频段幅值。结果 与对照组相比,SHRs肠壁血流灌注水平显著降低(P<0.001)。QDZJN和各剂量PDG治疗2个月后,SHRs肠壁血流灌注水平显著升高(P<0.001)。SHRs组肠壁微血流灌注一氧化氮(NO)依赖和非依赖性内皮细胞源频段振幅均显著低于对照组(P<0.01)。QDZJN和PDG治疗可显著提高内皮细胞源频段幅值(NO依赖：P<0.0001;NO非依赖：P<0.01)。结论 QDZJN和PDG治疗2个月可显著改善SHRs肠壁的微血流动力学和内皮功能障碍...     

降压中药JMT对自发性高血压大鼠微循环血流的影响

<正>目的:采用激光多普勒血流成像技术,测定自发性高血压大鼠(SHR)尾部微循环血流量,观察抗高血压中药济脉通(JMT)对高血压模型动物微循环血流的影响。方法:SHR大鼠分别灌胃给予降压中药JMT(10g/Kg、5/Kg、2.5g/Kg),连续8周。应用MoorFLPI散斑全帧实时扫描成像系统,分两个阶段进行测量:     

杜仲纯中药制剂和松脂醇二葡萄糖苷对自发性高血压大鼠肾脏微循环血流灌注及其频谱特征的影响

目的:探讨杜仲纯胶囊(EU)和松脂醇二葡萄糖苷(PDG)对自发性高血压大鼠(SHRs)肾脏微循环血流灌注及其频谱特征的影响。方法:将30只SHRs随机分为SHR组,SHR+EU组(6.00g/kg EU灌胃),SHR+PDG (L)组(8.75mg/kg PDG灌胃),SHR+PDG (M)组(17.50mg/kg PDG灌胃)和SHR+PDG (H)组(35.00mg/kg PDG灌胃),每组各6只。6只WKYs大鼠为Control组。各组进行灌胃给药2个月。手术暴露大鼠肾脏,利用激光多普勒血流探测仪和血流成像仪检测并比较各组大鼠肾脏微血流灌注水平和血流成像情况。采用小波变换比较各组肾脏内皮源性频段幅值。结果:与Control组相比,SHR组肾脏血流灌注水平显著降低(P<0.01)。治疗2个月后,与SHR组相比,SHR+EU组和SHR+PDG各剂量组肾脏血流灌注水平显著升高(P均<0.01)。与Control组相比,SHR组肾脏微血流灌注内皮细胞源频段振幅显著降低(P<0.01)。与SHR组比较,SHR+PDG(M)组、SHR+PDG(H)组中NO依赖和非依赖性内皮...     

急性心肌梗死大鼠联合应用阿司匹林和氯吡格雷致胃黏膜损伤及微循环改变

目的:观察阿司匹林和氯吡格雷联合应用对实验性急性心肌梗死(AMI)大鼠胃黏膜损伤的作用及微循环的影响。方法:SD大鼠60只,麻醉后结扎左冠状动脉前降支复制AMI模型,成模大鼠随机分为4组:(1)模型对照组(AMI组):蒸馏水(5ml/kg/天)灌胃;(2)阿司匹林组(AMI+阿司匹林组):阿司匹林(9mg/kg/天)灌胃;(3)氯吡格雷组(AMI+氯吡格雷组):氯吡格雷(6.75mg/kg/天)灌胃;(4)双联抗(阿司匹林和氯吡格雷)组(AMI+双联抗组):阿司匹林(9mg/kg/天)和氯吡格雷(6.75mg/kg/天)混悬液灌胃;另设假手术组(只穿刺不结扎冠脉,其余处理方法同模型对照组)作为对照。每组12只,雌雄各半。连续灌胃28天,次日麻醉后剖腹暴露胃黏膜,采用微循环观测系统观察恒温状态下各组大鼠胃黏膜微循环的毛细血管管径、数量、血流状态。同时取腹主动脉血,放射免疫法测定其6-酮-前列腺素F1α(6-keto-PGF_(1α))和血栓烷B_2(TXB_2)含量。之后处死大鼠摘取胃体,解剖显微镜下计数胃黏膜出血点并行Guth评分,HE染色观察胃组织病理学形态并行Whittle评分评价胃黏膜损伤程度。结果:与假手术组比较,AMI+阿司匹林组大鼠胃黏膜损伤程度明显,胃黏膜微循环受累,血管扩张,管壁变薄,排列紊乱(P<0.05);AMI+双联抗组胃黏膜损伤程度加重,胃黏膜微血管扩张、通透性增强,毛细血管交集数减少(P<0.05)。与假手术组比较,AMI组胃黏膜毛细血管数量增加(P<0.05)。与AMI组比较,AMI+阿司匹林组、AMI+双联抗组大鼠胃黏膜集合毛细血管管径显著增加(P<0.01),AMI+双联抗组毛细血管交集数减少(P<0.05)、毛细血管数量显著增加(P<0.01)。与假手术组比较,AMI组大鼠6-keto-PGF_(1α)水平显著下降(P<0.01),AMI+双联抗组、AMI+阿司匹林组和AMI+氯吡格雷组6-keto-PGF_(1α)和TXB_2水平均有不同程度降低(P<0.05,P<0.01)。与AMI组大鼠比较,给药各组6-keto-PGF_(1α)水平无明显变化(P>0.05),TXB_2水平明显降低(P<0.01)。结论:阿司匹林和氯吡格雷联用可加重AMI大鼠胃黏膜损伤,其机制可能与血液6-keto-PGF_(1α)与TXB_2水平降低和失衡,以及胃黏膜微循环障碍有关。     

葛根素对自发性高血压大鼠脑微循环的影响

目的:研究葛根素对于自发性高血压大鼠(SHR)脑微循环的影响及其与脑组织损伤的关系。方法:SHR随机分为葛根素治疗、溶剂对照(20%丙二醇)、阳性药物(尼莫地平)对照和空白对照等4组,连续观察14天,记录颈总动脉血压、软脑膜微循环和脑组织血流量变化,HE染色观察脑组织改变,Ⅷ因子染色测定微血管密度。结果:葛根素治疗14天后SHR血压降至正常范围内,软脑膜细动脉血管内径明显大于对照组,病理切片显示治疗组脑细动脉壁重塑,局部缺血明显减轻。结论:葛根素治疗可以扩张SHR脑细动脉、减轻高血压引起的微血管与脑组织损伤。     

自发性高血压大鼠肾脏和胰腺微循环血流灌注和自律运动变化

目的:观察自发性高血压大鼠(SHRs)肾脏和胰腺微循环血流灌注和自律运动变化。方法:市购8周龄SHRs(SHR组)和正常血压对照大鼠(WKY组)各6只,手术暴露肾脏和胰腺。采用双通道激光多普勒血流仪,多点观测其表面的微血流灌注和自律运动水平,并通过仪器自带分析软件的小波变换功能将微血流信号转换为二维频谱图和三维时频图像,比较两组大鼠肾脏、胰腺血流灌注水平、血流速度和血细胞浓度的变化特征及微血流灌注频谱中一氧化氮(NO)依赖与否的内皮细胞源频段幅值差异。结果:与WKY组相比,SHR组肾脏、胰腺微血流灌注水平均显著降低(P<0.05或P<0.01);同时呈现胰腺微血管自律运动节律紊乱。SHR组肾脏和胰腺微血管自律运动振幅和频率均显著降低(P<0.05或P<0.01)。SHR组肾脏和胰腺微血流灌注频谱中NO依赖与否的内皮细胞源频段振幅均显著低于WKY组(P<0.05或P<0.01)。结论:SHRs肾脏、胰腺微循环功能异常,表现为微血流灌注水平下降,微血管自律运动功能障碍。     

早期自发性高血压大鼠末梢微循环功能障碍和形态异常研究

目的:检测分析早期自发性高血压大鼠(SHR)末梢微循环功能障碍和形态变化及其与高血压的相关性.方法:选取8周龄SHR大鼠(SHR)组和正常血压WKY大鼠(对照组)各6只,先行复测收缩压、舒张压、平均动脉压和脉压差,再使用激光多普勒血流成像仪、高功率针式激光多普勒微循环检测仪及激光微循环形态和功能分析仪,分别检测其耳廊边...     

糖肾方对糖尿病肾病大鼠血液流变学及内皮细胞功能的影响

目的:观察糖肾方对糖尿病肾病(DN)大鼠血液流变学及内皮功能的影响。方法:采用高脂高糖饮食联合链脲佐菌素(STZ)诱导建立2型糖尿病合并DN大鼠模型,将建模成功大鼠随机分为DN模型组、糖肾方组和缬沙坦组,同时设正常对照组。药物干预12周后,全自动生化分析仪检测大鼠空腹血糖(FBG)、血清总胆固醇(TC)和甘油三酯(TG)水平;微循环血流仪检测肾微循环变化;血液流变仪检测血液流变学指标;ELISA技术检测尿白蛋白以及血清假血友病因子(v WF)和纤溶酶原激活物抑制剂-1(PAI-1)含量。结果:与正常对照组比较,DN模型组大鼠FBG、血清TC、TG和尿白蛋白明显升高(P0.05),血清v WF浓度升高(P0.01);与DN模型组比较,糖肾方组和缬沙坦组的FBG差异无统计学显著性,血清TC、TG和尿白蛋白明显降低(P0.05),v WF降低(P0.01)。与正常对照组比较,DN模型组的血浆黏度,卡松黏度,全血高切、中切黏度,红细胞聚集指数,刚性指数及电泳时间均显著升高,红细胞变形指数降低(P0.01)。与DN模型组比较,治疗组的血浆黏度,卡松黏度,全血高切、中切和低切黏度,红细胞聚集指数,刚性指数及电泳时间均明显降低(P0.01),红细胞变形指数升高,但无显著性差异。与正常对照组比较,DN模型组的肾皮质微循环血流量减少(P0.01)。与DN模型组比较,糖肾方组的肾皮质微循环明显增加(P0.05),缬沙坦组无明显改变。结论:糖肾方能降低实验性DN大鼠的血脂水平,改善血液流变性和内皮功能,从而改善微循环,对糖尿病肾脏具有一定的保护作用。     

Laser Doppler assessment of human colonic blood flow

We have used laser Doppler flowmetry to assess human colonic blood flow in patients with healthy mucosa and in those with active proctitis. We report a significant correlation between colonic serosal laser Doppler flux readings and serosal tissue oxygen tension (n = 45: r = 0.671: p = 0.001). Mucosal blood flow was measured using the PF309 probe of a Perimed Pf2b set on 12 kHz frequency shift. There was no significant difference throughout the colon in healthy patients. The flow in rectal mucosa was significantly higher in patients with proctocolitis than in normal mucosa.     

Laser Doppler assessment of human colonic blood flow

We have used laser Doppler flowmetry to assess human colonic blood flow in patients with healthy mucosa and in those with active proctitis. We report a significant correlation between colonic serosal laser Doppler flux readings and serosal tissue oxygen tension (n = 45: r = 0–671: p = 0–001). Mucosal blood flow was measured using the PF309 probe of a Perimed Pf2b set on 12 kHz frequency shift. There was no significant difference throughout the colon in healthy patients. The flow in rectal mucosa was significantly higher in patients with proctocolitis than in normal mucosa.     

Quantification of Weal Reactions with Laser Doppler Flowmetry

Skin-prick experiments with different concentrations of histamine (0.1 mg/ml, 1.0 mg/ml, 10.0 mg/ml) and isotonic NaCl solution for control were performed on the forearms of 10 healthy volunteers. Regular recordings of the cutaneous blood flow by laser Doppler flowmetry were performed in the centre of the weals and 5, 10, 15 and 20 mm from the centre representing the zone of perilesional flare. In the weal centre the blood flow was increased but neither different concentrations of histamine nor the control reaction could be differentiated. Recordings performed 5 and 10 mm from the centre showed, however, significant differences between the different solutions of histamine tested as well as the control reaction. The better differentiation was obtained 5 and 10 min after skin-prick. In conclusion, laser Doppler flowmetry with recordings of the perilesional flare is found useful for quantification of weal reactions.     

Laser Doppler measurement of blood velocity in microvessels

The application of laser Doppler velocimetry to the measurement of blood flow in narrow vessels is considered. Particular attention is paid to the effect of the finite laser probe volume (velocity gradient averaging). Successful in vitro results are reported but it is concluded that in vivo applications are likely to be confined to certain accessible vascular systems.     

Laser Doppler perfusion monitoring and imaging: novel approaches

Laser Doppler flowmetry (LDF) is a non invasive method enabling the monitoring of microvascular blood flow, a very important marker of tissue health. This article gives an overview on the concept of LDF for microvascular perfusion monitoring and imaging. It first describes the theoretical background of the technique. Then, the benefits of LDF signal processing are shown through clinical examples: use of time-frequency representations and wavelets. Afterwards, the paper introduces novel approaches of velocity components. For that purpose, a work providing the determination of the velocities relative contribution in physiologically relevant units (mm/s) is presented. Imaging perfusion is also reviewed through methods based on laser speckle. The most prominent disadvantage of the latter devices being the time needed to produce a perfusion image, solutions are proposed in the last part of the paper.     

Laser Doppler flowmetry: characteristics of a modified single-fibre technique

The single-fibre percutaneous laser Doppler technique has been used in previous studies of intramuscular blood flow. This method facilitates studies of blood flow in deep tissue volumes and minimises the tissue trauma. The technique has been further developed with the aim of improving the signal quality. This has been accomplished by modifying the geometry of the fibre tip. By melting the fibre core material, lenses of different shapes are formed. Flat, spherical and ‘pears-type tips have been manufactured and are evaluated theoretically and experimentally. The paraxial theory cannot accurately predict the position of zones of highest irradiance. Therefore, a ray-tracing program has been developed in the C language, by means of which some of the optical properties of the modified fibre tips can be simulated. Isoirradiance graphs and beam profiles are calculated for the three different fibre tips. Measured and calculated irradiance curves are used for evaluation of the properties of the ray-tracing model. The three types of fibre tips are also evaluated and compared in flow models. The sphere and pear-type probes show a higher flow sensitivity than the flat-end type. These improvements in flow sensitivity are interpreted as being related to the larger, strongly irradiated tissue volumes in front of the fibres. Intramuscular measurements with the pear-type probe show high sensitivities to induced blood flow changes.     

Measurements of enlarged blood pump models using Laser Doppler Anemometer

In an earlier study (Chua et al., 1998, 1999a), a 5:1 enlarged model of the Kyoto-NTN Magnetically Suspended Centrifugal Blood Pump (Akamatsu et al., 1995) with five different impeller blade profiles was designed and constructed. Their respective flow characteristics with respect to (1) the three different blade profile designs: forward, radial, and backward, (2) the number of blades used, and (3) the rotating speed were investigated. Among the five impeller designs, the results obtained suggested that impellers A and C designs should be adopted if higher head is required. Impellers A and C therefore were selected for the flow in between their blades to be measured using Laser Doppler Anemometer (LDA), so as to have a better understanding of the flow physics with respect to the design parameters.     

高血压微循环与体循环动力学临床研究系列——Ⅳ.利用激光多普勒血流计测评高血压患者反应性充血

在本实验中,我们利用激光多普勒血流计测量了25例高血压病人的手指皮肤阻断后反应性充血(PORH)及降压治疗对其影响,结果显示高血压病人(PORH)过程较正常人显著延长(P<0.05),而各流量参数在两组间则无显著差异,同时,我们发现,手腕及手指根部阻断时对于正常人,充血峰值的静息值百分比(PF％)和静息值(RF)之间存在着良好的相关性,而此种情况不存在于高血压病人,经降压治疗后,病人组反应性充血较前加快(P<0.05),此外,我们评价了年龄对正常人PORH的影响,发现较老年组PORH慢于较青年组(P<0.05)。     

内脏器官激光多谱勒血流灌注图象的显示

目的显示内脏器官血流灌注和分布的图象,探讨激光多谱勒血流成像技术在内脏微循环检测中的价值。方法应用激光多谱勒血流灌注成像仪(LDPI),对大鼠和家兔腹部脏器(肝、脾、肾、膀胱、胃、大小肠和肠系膜等)的血流灌注进行扫描和血流图显示。结果在体内脏器官显像清晰,肝、肾、脾、膀胱等微循环血流的整体分布较均匀;而胃、肠系膜、回盲肠等激光多谱勒血流图上大血流线状显示极其显著;大鼠与家兔两种动物之间相同脏器血流图上的差异并不明显。结论LDPI能将内脏器官的微循环状态以图象的形式显示,该新技术方法在检测内脏血流方面将有一定的应用价值。