Rod-shaped tubulated bodies in the endothelia of mesenteric arteries of hypertensive rats.

Weibel-Palade bodies were found in the endothelial cells of the mesenteric arteries of hypertensive rats. They were spherical or rod-shaped and bounded by a single membrane. They measured approximately 0.1 to 0.2 mu in diameter and up to 0.4 mu in length. The majority of them showed microtubular subunits, measuring about 200 A in width. These bodies are thought to originate from Golgi apparatus.     

烫伤大鼠脑底动脉壁内含内皮素-1能神经纤维的形态学变化

目的探讨烫(烧)伤损伤时大鼠脑血管内皮素-1能神经纤维分布与脑血管神经源性调节的关系,以及烫(烧)伤对脑血管损伤的影响。方法应用免疫组织化学技术观察烫(烧)伤大鼠脑底动脉(包括大脑前动脉、大脑中动脉、大脑后动脉和基底动脉)内皮素-1能神经纤维的分布。结果烫(烧)伤大鼠和正常大鼠脑底动脉均可见棕褐色的内皮素-1能免疫反应阳性神经纤维,似细线状,攀附于血管壁上,烫(烧)伤大鼠脑底动脉各主要分支内皮素-1能免疫反应阳性纤维密度较正常大鼠明显增加,纤维走行大多呈网状。结论烫(烧)伤可引起大鼠脑底动脉内皮素-1能免疫反应阳性神经纤维增加,增加的内皮素-1能神经纤维可能诱发脑血管痉挛和脑血液循环紊乱。提示内皮素-1能神经纤维在烫(烧)伤后在脑血管的神经源性调节中可能起重要的作用。     

Age-dependent increase of magnesium in the cerebral arteries of Thai

To elucidate compositional changes of the cerebral arteries with aging, the authors investigated age-related changes of elements in the cerebral arteries by inductively coupled plasma-atomic emission spectrometry (ICP-AES). After an ordinary dissection by medical students at Chiang Mai University was finished, the anterior, middle, and posterior cerebral arteries and the anterior and posterior communicating arteries were resected from the subjects. The subjects consisted of six men and seven women, ranging in age from 34 to 86 yr. The element content was determined by ICP-AES. It was found that the Mg content increased progressively with aging in all of the anterior, middle, and posterior cerebral arteries, but the Ca and P content did not increase significantly with aging in the anterior, middle, and posterior cerebral arteries, with the one exception being that the P content increased significantly in the posterior cerebral arteries with aging. The average content of Ca was higher in the order of the anterior communicating, anterior cerebral, posterior communicating, posterior cerebral, and middle cerebral arteries.     

Reiter株轴丝的分离及鉴定

本试验将Reiter株菌体超声破碎,上清液经离子交换柱层析及凝胶过滤,得到纯化轴丝.该轴丝经SOS-PAGE检测分析,所得结果与丹麦参考品一致,分子量在3万到4万之间,与文献报导相同.经电镜观察,产物均匀,未被杂质污染.按上述方法提取,3克湿重菌体最终可得2毫克左右轴丝蛋白,整个纯化过程在短时间内即可完成.     

Pathway of nerves with vasoactive intestinal polypeptide-like immunoreactivity to the major cerebral arteries of the rat

Summary The pathway of nerves with vasoactive intestinal polypeptide(VIP)-like immunoreactivity to the major cerebral arteries was studied in rats by means of the indirect immunofluorescent method. The fibers are densely distributed in the ethmoidal nerves and in the adventitia of both the external and internal ethmoidal arteries. Section of both ethmoidal nerves and external ethmoidal arteries before they enter the cranial cavity induced a marked reduction of VIP-like immunoreactive fibers in the walls of the vessels of the circle of Willis and its major branches. However, section of the external ethmoidal artery alone did not result in visible changes of the nerves around major cerebral arteries. The present study suggests that VIP-like immunoreactive fibers surrounding major cerebral arteries of the rat arise from fibers in the ethmoidal nerve showing immunoreactivity to VIP.     

开放翼腭动脉改良制备大鼠自体血栓脑梗死模型比较

目的：利用大鼠自体血栓,在开放翼腭动脉（ pterygopalatine artery,PPA）的情况下堵塞其大脑中动脉,造成大脑中动脉远端供血区的脑梗死模型。方法开放PPA,结扎颈外动脉（ external carotid artery,ECA）,将栓子通过颈总动脉（common carotid artery,CCA）血流将栓子冲大脑中动脉（middle cerebral artery,MCA）远端,造成梗死模型,按Bederson评分对其神经功能进行评分,TTC染色确定梗死体积。结果两手术组模型比较,评分均升高,且出现白色梗死区,经统计检测分析无明显差异。结论开放PPA大鼠自体血栓制造脑梗死模型可缩短手术时间,降低死亡率。     

Ultrastructural localization of nitric oxide synthase and endothelin in coronary and pulmonary arteries of newborn rats

This is the first report on the ultrastructural distribution of nitric oxide synthase and endothelin immunoreactivities in the coronary and pulmonary arteries of newborn Wistar rats. The distribution of nitric oxide synthase and endothelin was investigated using pre-embedding peroxidase-antiperoxidase immunocytochemistry. In both arteries examined, positive labelling for nitric oxide synthase was localized both in the endothelium and smooth muscle, whereas positive labelling for endothelin was localized in the endothelium exclusively. In the coronary artery, approximately 80% and 55% of the endothelial cells examined were positive for nitric oxide synthase and endothelin, respectively, whereas in the pulmonary artery, 77% and 60% of the endothelial cells were positive for nitric oxide synthase and endothelin, respectively. These findings indicate that nitric oxide synthase and endothelin are colocalized in some of the endothelial cells of the newborn rat. In the endothelium, nitric oxide synthase and endothelin immunoreactivities were distributed throughout the cell cytoplasm and in association with the membranes of intracellular organelles. In smooth muscle, a relationship of nitric oxide synthase immunoreactivity to endoplasmic reticulum was observed in the pulmonary artery. In summary, in the newborn rat, endothelial cells of the coronary and pulmonary artery are rich in nitric oxide synthase (neuronal isoform) and endothelin, and it is suggested therefore that they may be substantially involved in vasomotor control of the cardiac and pulmonary circulation during early stages of postnatal development.     

线栓法制备大鼠局灶性脑缺血模型手术操作的研究进展

线栓法大鼠大脑中动脉阻塞模型被普遍认为是标准的大鼠局灶性脑缺血动物模型。该模型在制备过程中,易受到多重因素的影响,其中手术操作对模型的成功制作起着关键作用。文章从大鼠颈部主要动脉的解剖位置、手术切口位置的选择、手术左右侧别的选择,以及插线位置的选择进行了综述。     

株纯化轴丝在梅毒血清学中初步应用

本试验以纯化的Reiter株轴丝为抗原,建立了AF—ELISA试验方法,用该方法检测了35份自家免疫病人血清、285份非梅毒病人血清及78份各期梅毒病人血清共398份血清,并同时与FTA-ABS等进行比较,结果AF-ELISA特异性为99.3%,FTA—ABS为99.7%,两者敏感性为100%.     

Calcium and phosphorus in aged human cerebral arteries

To elucidate the compositional changes of the cerebral arteries with aging, the authors investigated age-related changes of the calcium and phosphorus contents in the cerebral arteries by inductively coupled plasma-atomic emission spectrometry. The subjects consisted of 11 men and 5 women, ranging in age from 52 to 96 yr. The anterior, middle, and posterior cerebral arteries derived from the same subjects were used in the present study. It was found that there were no significant relationships between age and calcium or phosphorus content in the anterior, middle, and posterior cerebral arteries, indicating that the accumulation of calcium and phosphorus scarcely occurred in the anterior, middle, and posterior cerebral arteries with aging. It was examined whether there were relationships in the calcium and phosphorus contents among the anterior, middle, and posterior cerebral arteries. It was found that there was a significant relationship in both the contents of calcium and phosphorus between the middle and posterior cerebral arteries, but not between the anterior and middle cerebral arteries nor between the anterior and posterior cerebral arteries.     

Effect of ischemia on TBARS and lactate production in several cerebral regions of anaesthetised and awake rats

The premise of neuroprotective therapy for acute ischemic stroke is based upon the possibility to interfere with the cellular ischemic cascade, so the understanding of the mechanisms and consequences of cerebral ischemia is necessary. The relationship between lipid peroxidation and acidosis was investigated in several regions of rat brain following ischemia without reperfusion. Male Wistar rats (280-300 g) were anaesthetised (Ketalar 33 mg/kg and Rompun 6.66 mg/kg) or not and underwent a four-vessel occlusion for 5 minutes. Then, thiobarbituric acid-reactive substances (TBARS) and lactate levels were measured in different brain regions (cerebellum, bulb, striatum, hippocampus, cortex). Induction of ischemia by ligation of two common carotid arteries and two vertebral arteries resulted in a production of TBARS (40-120%, p < 0.05) and lactate (20-60%, p < 0.05) in all cerebral regions of awake rats, especially in striatum, suggesting a potential link between lipid peroxidation and acidosis. When ischemia was realised on anaesthetised animals, an increase of lactate levels (30-50%, p < 0.05) was shown in all brain regions but TBARS were produced only in striatum (82%, p < 0.05). These data showed the particular vulnerability of striatum to ischemia and the possible opposite effects of an anaesthesia.     

双侧迷走神经和颈总动脉联合切断法建立急性脑缺血大鼠模型

目的建立一种操作简单的急性脑缺血动物模型。方法取雄性Wistar大鼠40只,体重200～230g,手术前禁食12 h,自由饮水,随机分为对照组A、B、C组及模型D组,共4组,每组10只。即A组:假手术组,仅切开颈部两侧皮肤,分离双侧颈总动脉和迷走神经,不切断,然后缝合;B组:仅切断双侧颈部迷走神经;C组:结扎并切断双侧颈总动脉(CCA);D组:联合组,即结扎并切断双侧颈总动脉,同时切断双侧颈部迷走神经。观察各组大鼠手术后的脑缺血症状,记录各组大鼠在8h内的死亡情况,超过8h死亡的动物按8h计,计算死亡率和死亡时间。结果 A组大鼠没有脑缺血症状,无死亡;B组大鼠无脑缺血症状,呼吸变慢变深,心率血压上升,但无死亡;C组大鼠部分出现脑缺血症状,眼睑下垂,活动能力低下,自发运动减少,有些大鼠术后自发运动增加,在8 h内无死亡;D组大鼠大多数出现较为明显的脑缺血症状,在8 h内全部死亡。结论采取同时结扎并切断大鼠双侧颈总动脉和双侧颈部迷走神经的方法,可以建立急性脑缺血大鼠动物模型,此方法具有手术简单,成功率高,术后动物缓慢死亡的特点。     

Increased density of fluorescent adrenergic fibers around the middle cerebral arteries of stroke-prone spontaneously hypertensive rats

The distribution of fluorescent adrenergic nerve fibers in the proximal portion (horizontal segment, Hs) and the three distal portions (major branches) of the middle cerebral arteries (MCA) was examined in stroke-prone spontaneously hypertensive rats (SHRSP) aged 10, 30, 60, 90, and 180 days, by the glyoxylic acid method. The results were compared with those in agematched normotensive Wistar Kyoto (WKY) rats. While the distribution pattern of fluorescent nerve fibers in the proximal portion of WKY rats changed from a straight linear arrangement at 10 and 30 days of age to a network-like arrangement after 60 days, those from SHRSP showed a constant meshwork pattern throughout the entire examination period. In the distal portions of the MCA of both SHRSP and WKY rats at all ages examined, fluorescent nerve fibers formed a coarse network. The distribution densities of adrenergic nerve fibers in the proximal and distal portions of the MCA of SHRSP were significantly higher (P<0.01 and 0.05) than those of WKY rats at all ages examined, except in the proximal portion at 90 and 180 days of age. The difference in nerve fiber density between SHRSP and WKY rats reached a peak at 30 days of age in both proximal and distal portions, and then gradually decreased with age. The present study suggests that sympathetic hyperinnervation is an important factor in the development of hypertension, and is involved in its maintenance in SHRSP.     

Bradykinin dilates rat middle cerebral artery and its large branches via endothelial B2 receptors and release of nitric oxide

Ring segments of rat middle cerebral artery (MCA) were prepared for measurement of isometric force and precontracted with 10⁻⁴ M uridine triphosphate (UTP). Concentration-effect curves (CEC) were constructed for bradykinin (BK, 10⁻⁸–10⁻⁵ M) in segments with functionally intect (E+) or denuded (E−) endothelium. E− segments did not dilate to BK. The BK receptor was characterized by application of specific B₁ or B₂ antagonists [des-Arg⁹-Leu⁸] BK (10⁻⁵ M) and [ -Arg⁰-Hyp³-Thi⁵- -Tic⁷-Oic⁸] BK (HOE140,3 × 10⁻⁷ M), respectively, or B₁ agonist [des-Arg⁹] BK (10⁻⁸–10⁻⁴ M). Involvement of nitric oxide (NO) was tested with N^G-nitro- -arginine (LNNA, 10⁻⁴ M). BK induced concentration-dependent relaxation with a maximal effect (E_max) of 40.86 ± 1.50% at 10⁻⁶ M and a pD₂ (−log₁₀ EC₅₀) of 6.818 ± 0.044. This relaxation could be prevented with HOE140 or LNNA, but was not influenced by [des-Arg⁹-Leu⁸] BK. [des-Arg⁹] BK did not induce any effect. These results demonstrate that BK induced relaxation via endothelial B₂ receptors and release of NO in isolated rat MCA.     

Interendothelial junctions of cardiac capillaries in rats: their structure and permeability properties

Summary The isolated perfused heart model was used to examine the structure of rat cardiac capillaries and their permeability to macromolecules of various sizes. Haemoglobin (diameter 6.4 nm) and catalase (10.4 nm) did not cross the endothelium but remained on the luminal side. Cytochrome C (3 nm) and horseradish peroxidase (6 nm) both crossed the endothelium to the subendothelial space and filled the caveolae on the abluminal side as well as the entire length of the lateral intercellular spaces. The membranes of the endothelial cells are separated by an intercellular gap of mean width 18.2 nm. At one or more zonular regions within each lateral intercellular space the two membranes approach each other more closely and frequently appear to fuse. However, tilting the specimen shows that, in these regions, there is a gap of mean width 5.4 nm (in lanthanum- and tannic acid-treated tissue, 3.8 nm in ferrocyanide-treated tissue) between the membranes. We conclude that these narrow regions sieve macromolecules on the basis of size although other factors may determine their permeability properties.     

Morphological changes in cerebral vascular smooth muscle cells in stroke-prone spontaneously hypertensive rats (SHRSP)

The ultrastructure of the vascular smooth muscle cells of the middle cerebral artery in 6-month-old male stroke-prone spontaneously hypertensive rats (SHRSP) was studied by scanning (SEM) and transmission electron microscopy (TEM) and compared with that of age-matched normotensive Wistar Kyoto rats (WKY). Although the smooth muscle cells of WKY rats by SEM had a typical spindle shape and smooth surface texture, those of SHRSP were structurally modified by numerous surface invaginations and projections, bearing some structural resemblance to the myotendinous junction of skeletal muscle. Structural modifications affected more than half the surface of medial smooth muscle cells in SHRSP, but less than 0.6% of the surface of these cells in WKY rats. About 10% of medial smooth muscle cells were necrotic in SHRSP, but no necrotic cells were identified in WKY rats. By TEM, smooth muscle cells in SHRSP were shown to be irregular in profile with deep indentations of the plasma membrane and were surrounded by many layers of basal laminalike material. The present study suggests that most smooth muscle cells in the middle cerebral artery of SHRSP may be modified to adapt to chronic hypertension by increasing the junctional area between muscle cells and connective tissue and that some cells may undergo necrosis.     

Anti-heat shock protein 70 autoantibody epitope changes and BD091 promotes atherosclerosis in rats

It has been previously reported that the plasma levels of autoantibodies against heat shock protein 70 (HSP70) are elevated in atherosclerosis. The aim of the present study was to elucidate whether anti-HSP70 antibodies are involved in the pathogenesis of atherosclerosis. To determine this, we chose rats as an atherosclerosis model. Titers of plasma anti-HSP70 autoantibody were determined by ELISA. After the intravenous administration of antibody into the tail, the damaged areas of aorta were stained with Evans Blue, atheromatous plaque were stained by Oil Red O, and then they were measured and quantified with AxioVision computer software. The number of macrophages ( $ {{\hbox{M}}_\Phi } $ ), smooth muscle cells (SMCs), and T cells were determined by immunocytochemistry. The level of anti-HSP70 IgG1 antibody was apparently increased in the AS group at the tenth week, and one hybridoma of HSP70 antibody (BD091, IgG1, recognizing C-terminal) had the same binding epitope as plasma anti-HSP70 autoantibodies. After intravenous administration, the lesion area of aorta with BD091 was significantly larger than those of IgG^mouse and SPA-810. Moreover, injection of BD091 resulted in significant endothelium damage, followed by a greater accumulation of $ {{\hbox{M}}_\Phi } $ , T cells, and SMCs in lesions than in the control. In conclusion, BD091 reaction with HSP70 expressed on arterial endothelial cells inducing endothelium damage triggers the inflammatory response in the vessel wall that accelerates atherosclerosis in rats. BD091 shares the same binding epitope with HSP70 autoantibodies. These data indicated that a specific epitope of anti-HSP70 autoantibody participated in the pathogenesis of atherosclerosis.     

The effect of ozagrel sodium on photochemical thrombosis in rat: therapeutic window and combined therapy with heparin sodium

The therapeutic efficacy of ozagrel sodium (ozagrel), alone and in combination with heparin, and its therapeutic time window were studied in a photochemically induced thrombotic cerebral infarction rat model. Cerebral artery thrombosis was induced by irradiating the brain with green light through intact skull using rose bengal as the photosensitizing dye. One set of animals was treated immediately after thrombosis with (1) vehicle, (2) 10 mg/kg ozagrel in saline, intravenously (i.v.), (3) 150 U/kg unfractioned heparin, subcutaneously (s.c.), or (4) ozagrel, i.v. plus heparin, s.c. Infarct volume was significantly smaller and edema was reduced in the ozagrel-treated groups compared to the vehicle-treated group; heparin did not convey additional benefit. In another set of animals, rats were given either vehicle or 10 mg/kg ozagrel in saline, i.v., 60 min or 120 min after induction of thrombosis. Ozagrel reduced infarct volume, but its effect diminished with delayed administration. The therapeutic window was determined to be less than 60 minutes after induction of thrombosis.