新发现的血管外膜源舒张因子

血管内皮依赖的舒张因子(EDRF)即一氧化氮(NO)的发现,对血管生物学的发展和血管性疾病的研究带来巨大的影响和推动,并因此而荣获诺贝尔奖。既往认为,血管外膜的功能是对血管起保护、支撑和营养作用。近年的研究发现,外膜还可以分泌多种血管活性物质,以旁分泌的方式调节血管的舒缩功能及结构变化。Matthias Lohn等灌注离体大鼠主动脉血管环,     

兔颈动、静脉移植桥血管狭窄模型的建立及其电镜观察

目的：建立兔颈动、静脉移植血管桥动物模型,观察移植桥血管内膜增生和狭窄的电镜下表现。方法：通过兔双侧颈动脉进行动脉桥和静脉桥的移植,形成双侧移植血管桥再狭窄动物模型。在第8周施行血管桥移植手术的同时留取右侧颈动静脉标本作为对照血管,再分别于第12周、16周和第20周分别处死模型兔,采集移植桥血管标本,在光镜下测量其内膜厚度、面积、狭窄度,并进行电镜观察。结果：颈动脉和颈静脉桥移植后,随着时间的延长,桥血管的出现平滑肌迁移,脂质沉积,内膜增生,血管狭窄等改变,且以静脉桥血管的病理改变更为明显。结论：在兔形成动脉粥样硬化病变基础上,进行双侧颈动脉血管桥的移植,建立兔双侧颈动脉移植血管桥再狭窄动物模型,有利于设立自身对照,研究术后动静脉桥再狭窄差异机制;建立动、静脉桥后,位于血管中膜的平滑肌细胞出现向血管内膜迁移现象,说明中膜平滑肌细胞迁移进入内膜导致新内膜形成是血管再狭窄的重要环节。     

兔颈动、静脉移植桥血管狭窄模型的建立及其电镜观察

目的:建立兔颈动、静脉移植血管桥动物模型,观察移植桥血管内膜增生和狭窄的电镜下表现。方法:通过兔双侧颈动脉进行动脉桥和静脉桥的移植,形成双侧移植血管桥再狭窄动物模型。在第8周施行血管桥移植手术的同时留取右侧颈动静脉标本作为对照血管,再分别于第12周、16周和第20周分别处死模型兔,采集移植桥血管标本,在光镜下测量其内膜厚度、面积、狭窄度,并进行电镜观察。结果:颈动脉和颈静脉桥移植后,随着时间的延长,桥血管的出现平滑肌迁移,脂质沉积,内膜增生,血管狭窄等改变,且以静脉桥血管的病理改变更为明显。结论:在兔形成动脉粥样硬化病变基础上,进行双侧颈动脉血管桥的移植,建立兔双侧颈动脉移植血管桥再狭窄动物模型,有利于设立自身对照,研究术后动静脉桥再狭窄差异机制;建立动、静脉桥后,位于血管中膜的平滑肌细胞出现向血管内膜迁移现象,说明中膜平滑肌细胞迁移进入内膜导致新内膜形成是血管再狭窄的重要环节。     

SM22α对血管平滑肌细胞骨架及收缩功能的影响

SM22α(smooth muscle 22 alpha,SM22α)是血管平滑肌细胞(vascular smooth muscle cells,VSMC)的标志蛋白,为了探讨该蛋白与VSMC表型和功能的关系,利用血清饥饿法诱导VSMC由合成型向收缩型转变,用RT—PCR对不同表型VSMC的SM22α表达活性进行检测,并通过转染反义SM22α表达载体,观察SM22α表达对VSMC细胞骨架和收缩功能的影响。结果显示,在VSMC由合成型逆转为收缩型的过程中,SM22α和平滑肌α-肌动蛋白(smooth muscle α—actin,SMα—actin)的表达分别被显诱导和轻度上调,与此同时,细胞骨架由稀疏的网格状变成均匀、致密的束状,VSMC重新获得收缩功能。用反义SM22α抑制该基因表达后,血清饥饿诱导的VSMC细胞骨架重构受阻,乙酰胆碱刺激引发的细胞收缩消失。结果提示,SM22α参与VSMC细胞骨架的构成及调节细胞的收缩功能,对维持VSMC处于收缩表型具有重要作用。     

小鼠血管内膜增生模型的建立

该文主要介绍一种可以在小鼠中有效地诱导血管内膜增生的方法。该方法使用硅胶管嵌套小鼠股动脉,造成股动脉血管内膜增生。病理切片观察表明,实验组小鼠股动脉与假手术组相比出现了内膜层和中膜层不规则增厚,管腔狭窄,细胞排列紊乱,内膜炎症细胞浸润等。免疫组织化学染色显示内膜增生部分是由于血管平滑肌细胞（VsMCs）增生和基质积聚所致。该模型的建立对动脉粥样硬化病理机制研究和治疗药物的研发具有重要意义。     

球囊拉伤兔髂动脉内膜的组织形态变化的研究

球囊拉伤血管内膜是目前研究血管内膜增厚,管腔狭窄的方便而切实的模型。因此,对其演变过程的形态学研究是必要的。方法：应用ＰＴＣＡ球囊导管拉伤兔髂动脉,用光镜、电镜和扫描电镜观察拉伤后不同时间的动态变化。结果：拉伤后各期变化不同,拉伤后１周内,以血细胞沉积为主,１周后,内膜开始增生,２至４周增生最快。增生活跃的内膜平滑肌细胞来自活化的中膜平滑肌细胞。内皮细胞增生极慢,４周内未见内膜内皮化,呈虫食样改变。结论：球囊拉伤兔髂动脉内膜,可引起血栓形成,炎症反应,平滑肌细胞增生和细胞外基质堆积,导致血管腔狭窄。此研究为今后的工作提供一定的客观依据     

吸烟对大鼠肺循环血流动力学及肺血管反应性的影响

本实验用大鼠29只,进行人工通气吸入烟气,初步探讨了吸烟对肺循环的影响。其中7只观察了吸烟对肺循环血流动力学的直接影响,结果表明,吸烟可致右心室收缩压、心输出量下降及心率减慢,肺循环阻力无明显改变。观察22只大鼠吸烟后缺氧所致肺循环血流动力学变化,结果表明,吸烟可使缺氧性肺血管反应降低,而且发生在肺循环血流动力学变化之前。     

用差异显示PCR法筛选与血管外膜细胞表型转化相关的基因

为筛选血管外膜成纤维细胞（adventitial fibroblast,AF）与肌成纤维细胞（myofibroblast,MF）间表型转化有关的基因,实验建立了大鼠胸主动脉AF和MF两种细胞模型,用差异显示聚合酶链反应（DD－PCR）技术获得表达差异片段,对差异片段进行克隆和测序分析,并用定量PCR和Northern blot对差别显示结果进行验证。用反义核酸转染技术观察骨桥蛋白（osteopontin,OPN）对AF迁移的影响。结果表明,两种表型细胞存在明显的基因表达差异,其中一个在MF下调的差异片段与GenBank中NADH脱氢酶亚单位5（NADH dehydrogenase subunit 5,Nd5）基因高度同源。另一个在MF上调的差异片段与OPN基因同源。上述差异表达结果被定量PCR及Northern blot证实。此外还有4个表达序列标志（expressed sequence-tag,EST）在GenBank中未查到同源序列。反义OPN寡脱氧核甘酸可抑制AF的迁移活动。结果提示,AF转化为MF可能与ND5基因下调、OPN上调及其它未知基因的表达改变有关。应用反义技术适度抑制OPN表达在防治血管重塑中具有重要作用。     

Effect of removal of adventitia on vascular smooth muscle contraction and relaxation

The aim of the present study was to determine whether the adventitia of large arteries modulates vascular function. We developed a method to obtain functional vascular rings devoid of adventitia. Carotid and iliac arteries from 3-mo-old Sprague-Dawley rats were denuded from adventitia after treatment with collagenase followed by gentle peeling. Adventitia removal and integrity of the media was demonstrated by optical and confocal microscopy. Arterial rings with or without adventitia and with or without endothelium were mounted in an organ bath for isometric tension recording. Responses to 75 mM KCl or norepinephrine (0.1 nM-1 microM) were significantly reduced in segments without adventitia. Acetylcholine-induced relaxation (0.1 microM-0.1 mM) was enhanced in arteries without adventitia, whereas sodium nitroprusside-induced responses were not modified. These results demonstrate that the combination of stripping with a previous collagenase treatment allows us to obtain functional rings devoid of adventitia and that this layer plays a role in contractile capacity and in endothelium-modulated responses.     

Meningeal arterial patterns in great apes: Implications for hominid vascular evolution

Arterial meningeal patterns were observed for 100 hemispheres from great ape endocasts (Pan paniscus, Pan troglodytes, Gorilla gorilla, and Pongo pygmaeus). Eight patterns emerged based on the relative contributions to the walls and dura mater of the middle part of the braincase of meningeal arteries that stem from two sources. These arteries enter the braincase through either the orbit (delivering blood from the internal carotid artery) or through the base of the middle cranial fossa (via the middle meningeal artery whose blood comes from the external carotid artery). The three genera of apes manifest different frequencies of the eight, patterns, with orangutans highly dependent on orbital meningeal arteries at one extreme, and chimpanzees showing the greatest reliance on the middle meningeal artery at the other. As was the case in an earlier study of rhesus monkeys, there is a trend across the two genera of African apes for increased mean cranial capacity to be associated with increased reliance on the internal carotid artery for supplying the middle portion of the braincase. However, unlike the case for macaques, this trend does not reach statistical significance in African apes. Because it is rare for humans to manifest significant arterial contributions from the orbit to the middle cranial fossa, the comparative data on monkeys, apes, and humans suggest that, during the course of vascular evolution in Homo, the middle meningeal artery eventually took over supply of the entire middle cranial fossa. This hypothesis should be tested in the hominid fossil record. Earlier work on meningeal arterial patterns in apes has traditionally relied on Adachi's system that was determined from humans and focuses on the origin of the middle branch of the middle meningeal artery. As a result, the extensive orbital contributions to the middle portion of the braincase that characterize apes were not recognized and the eight patterns described in this paper were often erroneously assigned to the three patterns that adequately describe only humans. Adachi's system should therefore be abandoned for nonhuman primates and early hominids. A correct understanding of meningeal arterial evolution cannot be achieved until the orbital contributions to the meningeal arteries are recognized and incorporated into an evolutionary study that spans from apes to fossil hominids to living people. © 1993 Wiley-Liss, Inc.     

Effect of L-arginine on pulmonary artery smooth muscle cell apoptosis in rats with hypoxic pulmonary vascular structural remodeling

This study investigated the effect of L-arginine (L-Arg) on the apoptosis of pulmonary arterysmooth muscle cells (PASMC) in rats with hypoxic pulmonary vascular structural remodeling,and itsmechanisms.Seventeen Wistar rats were randomly divided into a control group (n=5),a hypoxia group(n=7),and a hypoxia L-Arg group (n=5).The morphologic changes of lung tissues were observed underoptical microscope.Using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay,the apoptosis of PASMC was examined.Fas expression in PASMC wasexamined using immunohistochemistry.The results showed that the percentage of muscularized artery insmall pulmonary vessels,and the relative medial thickness and relative medial area of the small and medianpulmonary muscularized arteries in the hypoxic group were all significantly increased.Pulmonary vascularstructural remodeling developed after hypoxia.Apoptotic smooth muscle cells of the small and median pul-monary arteries in the hypoxia group were significantly less than those in the control group.After 14 d ofhypoxia,Fas expression by smooth muscle cells of median and small pulmonary arteries was significantlyinhibited.L-Arg significantly inhibited hypoxic pulmonary vascular structural remodeling in association withan augmentation of apoptosis of smooth muscle cells as well as Fas expression in PASMC.These resultsshowed that L-Arg could play an important role in attenuating hypoxic pulmonary vascular structural remod-eling by upregulating Fas expression in PASMC,thus promoting the apoptosis of PASMC.     

Effects of papaya seed extract and benzyl isothiocyanate on vascular contraction

To investigate their potentially toxic effects on mammalian vascular smooth muscle, pentane extracts of papaya seeds and the chief active ingredient in the extracts, benzyl isothiocyanate (BITC), were tested for their effects on the contraction of strips of dog carotid artery. BITC and the papaya seed extract caused relaxation when added to tissue strips that had been pre-contracted with phenylephrine (PE). Incubation of the tissue with papaya seed extract or BITC caused inhibition of contraction when the strips were subsequently contracted with KCl or PE. This relaxation and inhibition of contraction did not appear to be endothelium-dependent, as endothelium-denuded rings showed the same degree of relaxation or inhibition of contraction in response to the preparations/drugs as those with the endothelium intact. The effects of both BITC and the extract were irreversible, i.e., the tissue did not recover to normal contractile ability after extensive washing. Exposure of the tissue to the papaya seed extract caused slower relaxation of the tissue, compared to controls, both after contraction with PE and subsequent addition of carbachol (CCh), and after contraction with KCl and then washing. Calcium imaging studies using cultured endothelial cells showed strong influxes of Ca2+ into the cells in response to addition of the papaya seed extract. We conclude that these extracts, when present in high concentration, are cytotoxic by increasing the membrane permeability to Ca2+, and that the vascular effects of papaya seed extracts are consistent with the notion that BITC is the chief bio-active ingredient.     

外源性硫化氢对家兔内毒素休克诱发肺动脉高压的干预作用

目的:探讨外源性硫化氢(H₂S)对家兔内毒素休克(ES)诱发肺动脉血管反应性的影响。方法:本实验采用家兔经颈静脉注射脂多糖(LPS,8 mg/0.8 ml/kg)复制家兔ES模型,并提前15 min腹腔注射H₂S供体硫氢化钠(NaHS,28μmol/kg)进行干预。随机将新西兰大耳白家兔分为4组(n=8):溶剂对照组、LPS组、LPS+NaHS组和NaHS组。监测平均动脉压(MAP)和平均肺动脉压(MPAP)的变化;应用血管环张力测定技术,检测各组家兔离体肺动脉张力变化;应用光镜和扫描电镜分别观察肺动脉管壁结构及肺动脉内皮细胞超微结构变化。结果:(1)注射LPS后家兔出现MAP降低、MPAP升高,成功复制家兔ES模型;与LPS组相比,LPS+NaHS组家兔MPAP在各个时间点均显著降低(P均﹤0.05);(2)与正常对照组相比,LPS组家兔肺动脉对苯肾上腺素(PE)的收缩反应增强,对乙酰胆碱(ACh)的舒张反应降低(P均﹤0.01);与LPS组相比,LPS+NaHS组家兔肺动脉对PE的收缩反应降低,而对ACh的舒张反应增强(P均﹤0.05)。...     

Effect of vasoconstriction on longitudinal distribution of pulmonary vascular pressure and volume

We used an improved version of the low-viscosity bolus method to evaluate longitudinal (arterial-to-venous) differences in the sensitivity of the dog lung lobe vasculature to selected vasoconstrictor stimuli, including hypoxia, and serotonin, histamine, and norepinephrine infusions. This method revealed a bimodal distribution of local vascular resistance vs. cumulative vascular volume under the zone 3 conditions studied. Our interpretation of the two modes of relatively high resistance is that they correspond to high resistance per unit volume segments of the arteries and veins upstream and downstream from the relatively low resistance per unit volume capillary bed. Thus an increase in the height of the upstream and downstream modes of the resistance distribution suggests constriction in small arteries and veins, respectively. Horizontal displacement of the modes along the cumulative volume axis suggests changes in the distribution of volume among the arteries, veins, and capillary bed. By use of these criteria, the results are consistent with the concept that each of the vasoconstrictor stimuli studied had a different longitudinal response pattern. Hypoxia constricted mainly small arteries, whereas serotonin constricted small and large arteries. Histamine constricted large and small veins, and norepinephrine constricted large and small veins and arteries.     

Effect of 0.25 T static magnetic field on microcirculation in rabbits

We showed previously in rabbits that 0.2 and 0.35 T static magnetic field (SMF) modulated systemic hemodynamics by arterial baroreceptors. We now have measured the effect of 0.25 T SMF on microcirculation within cutaneous tissue of the rabbit ear lobe by the rabbit ear chamber (REC) method. Forty experimental runs (20 controls and 20 SMF) were carried out in eight different rabbits with an equal number of control and SMF experiments on each individual. Rabbits were sedated by pentobarbital sodium (5 mg/kg/h, i.v.) during the entire 80 min experiment. SMF was generated by four neodium-iron-boron alloy (Nd2-Fe14-B) magnets (15 x 25 x 30 mm, Neomax, PIP - Tokyo Co., Ltd., Tokyo, Japan), positioned around the REC on the observing stage of an optical microscope. The direct intravital microscopic observation of the rabbit's ear microvascular net, along with simultaneous blood flow measurement by microphotoelectric plethysmography (MPPG), were performed PRE (20 min, baseline), DURING (40 min), and POST (20 min) magnetic field exposure. The control experiments were performed under the same conditions and according to the same time course, but without magnetic field. Data were analyzed comparing MPPG values and percent change from baseline in the same series, and between corresponding sections of control and SMF runs. In contrast to control series (100+/-0.0%-90.0+/-5.4%-87.7+/-7.1%, PRE-EXPOSURE-POST), after magnetic field exposure we observed increased blood flow (100+/-0.0%-117.8+/-9.6%*-113.8+/-14.0%, *P<0.05) which gradually decreased after exposure cessation. We propose that long exposure of a high level nonuniform SMF probably modifies microcirculatory homeostasis through modulation of the local release of endothelial neurohumoral and paracrine factors that act directly on the smooth muscle of the vascular wall, presumably by affecting ion channels or second messenger systems.     

Hippo信号通路在颈动脉结扎所致大鼠动脉血管重构中的表达及其意义

目的:在建立大鼠血管重构模型基础上探讨Hippo信号通路在该模型中的表达及意义。方法:模型组(n=40)经颈部正中切口游离出左侧颈总动脉,用6-0不可吸收线在尽量靠近近心端处结扎,完全阻断血流。对照组(n=20)仅将手术线穿过颈总动脉而不结扎,闭合切合。14 d后处死所有动物,经原手术路径分离颈总动脉,收集结扎处至远心端的动脉。用HE以及MASSON染色观察血管形态以及纤维化,免疫组织化学染色法检测颈动脉中α-肌动蛋白(α-MSA)和增殖细胞核抗原(PCNA)的表达,Western blot检测yes相关蛋白(YAP),PDZ结合基序的转录辅激活子(TAZ),TEAD1,Bax,Bcl-2的表达。结果:与对照组相比,造模组HE染色提示血管重构明显,新生内膜/中膜比例明显增加,MASSON染色提示纤维化明显增加;免疫组织化学染色法提示造模组血管α-MSA及PCNA表达明显增加; Western blot提示造模组血管YAP,TAZ,TEAD1,Bcl-2表达增加,而Bax表达降低,Bax/Bcl-2蛋白比例明显降低。结论:本研究成功建立颈动脉结扎介导的大鼠血管重构模型,另外证明Hippo信号通路在颈动脉结扎介导的大鼠血管重构模型中明显激活,以及可能介导增殖、凋亡相关的Bax/Bcl-2比值的改变,进而参与平滑肌细胞增殖促进血管重构。     

Regulation of tyrosine phosphatases in the adventitia during vascular remodelling

Protein tyrosine phosphatases (PTPs) are regulators of growth factor signalling in vascular remodelling. The aim of this study was to evaluate PTP expression in the context of PDGF-signalling in the adventitia after angioplasty. Utilising a rat carotid artery model, the adventitial layers of injured and non-injured vessels were laser microdissected. The mRNA expression of the PDGF β-receptor, the ligands PDGF-A/B/C/D and the receptor-antagonising PTPs (DEP-1, TC-PTP, SHP-2, PTP1B) were determined and correlated to vascular morphometrics, proliferation markers and PDGF β-receptor phosphorylation. The levels of the PDGF β-receptor, PDGF-C and PDGF-D were upregulated concurrently with the antagonising PTPs DEP-1 and TC-PTP at day 8, and normalised at day 14 after vessel injury. Although the proliferation parameters were time-dependently altered in the adventitial layer, the phosphorylation of the PDGF β-receptor remained unchanged. The expression dynamics of specific PTPs indicate a regulatory role of PDGF-signalling also in the adventitia during vascular remodelling.     

Effect of a linseed diet on lipogenesis, fatty acid composition and stearoyl-CoA-desaturase in rabbits

The aim of the study was to examine the effect of a linseed diet on meat quality and on lipogenesis in rabbits. Twelve rabbits were fed a control or a linseed diet. There was no diet effect on growth, food consumption, carcass characteristics and meat ultimate pH and colour. Feeding the linseed diet increased the n-3 polyunsaturated fatty acids (PUFA) levels in perirenal and interscapular fats, in the Longissimus dorsi muscle and in the liver. The linseed diet produced lower linoleic acid/α-linolenic acid ratios in adipose tissues and in the Longissimus dorsi muscle, but not in the liver. Diet did not affect lipogenic enzyme activities in the Longissimus dorsi muscle, whereas the linseed diet decreased the lipogenic potential in perirenal and interscapular fats, and in the liver. Feeding rabbits with a high n-3 PUFA diet led to a decrease in the oxidative stability of perirenal fat and the Longissimus dorsi muscle, and to an inhibition of stearoyl-CoA-desaturase activity in liver and in adipose tissues, but not in muscle.