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1.
Neurogenic inflammation of the dura mater encephali has been suggested to play an important role in the pathophysiology of headaches. Although functional studies using extravasation techniques indicate an enhanced permeability of blood vessels after chemical or electrical stimulation of C-fibres supplying the dura mater, histological demonstration of leaky blood vessels is still a problem. We used the vascular labelling method combined with i.v. injection of colloidal silver solution to test the permeability increasing effect of intravenous administration of substance P, topical application of mustard oil or acidic phosphate buffer and local electrical stimulation of the exposed dura mater. Histological characteristics of increased vascular permeability were observed exclusively after mustard oil and acidic phosphate buffer. This observation may indicate different mechanisms of increased vascular permeability involving pinocytosis and formation of interendothelial gaps selectively visualized by the vascular labelling method. 相似文献
2.
剪切力对动脉血管平滑肌细胞增殖分化的影响 总被引：2，自引：0，他引：2
目的：探讨剪切力作用下人脐动脉血管平滑肌细胞的增殖分化的变化。方法：利用改进的灌流及流动培养装置,通过蠕动泵提供稳定的剪切力,同时提供静态培养所需的其它条件,建立体外人脐动脉血管平滑肌流动培养模型。分别对体外培养的人脐动脉血管平滑肌细胞加载3,4,5,8,10dyn／cm^2的定长流剪切力24h,同时以静态培养的细胞为对照组。相差倒置显微镜观察玻片上细胞的数量并作细胞计数,α—actin免疫组化染色．结果：细胞记数显示,同对照组相比,七刀应力各组的增殖能力都有所下降,5dyn／cm^2切应力作用下,细胞增殖缓慢最为明显．α—actin免疫组化染色提示切应力各组的分化程度较对照组高。结论：初步研究表明,剪切力对人脐动脉血管平滑肌细胞的增殖有一定抑制作用,并且可能促进了细胞的分化。 相似文献
3.
Norma Risler Claudia Castro Montserrat Cruzado Susana González Roberto Miatello 《Biocell》2003,27(2):189-196
Remodeling of large and small arteries contributes to the development and complications of hypertension. Artery structural changes in chronic sustained hypertension include vascular smooth muscle cells (VSMC) proliferation and extracellular matrix (ECM) modifications. Extracellular constituents such as proteoglycans (PGs), may modulate vascular stiffness and VSMC growth and differentiation. We examined the effect of growth factors on secreted and membrane-bound PGs synthesis by cultured aortic smooth muscle cells (SMC) from 12- to 14- week-old spontaneously hypertensive rats (SHR) and age-matched Wistar rats. After stimulation with platelet-derived growth factor (PDGF-BB), 10% fetal calf serum (FCS) or 0.1% FCS as control, PGs synthesis (dpm/ng DNA) was evaluated in the medium (M-ECM) and in the cell layer (PECM) by a double-isotopic label method using both [3H]-glucosamine and [35S]-sodium sulfate which are incorporated into all complex carbohydrates or only into sulfated dysaccharides, respectively. Data are presented as percent of the control (0.1% FCS). SHR VSMC displayed a significantly greater synthesis of MECM [3H]-PGs than Wistar rat cells, with both treatments, but no differences in M-ECM [35S] uptake were found in any case. In the P-ECM, both PDGF-BB and 10% FCS produced a greater effect on [3H]-PGs and sulfated PGs synthesis in VSMC from SHR. An important change seen in SHR cells was a significant decreased sulfation, assesed by [35S]/[ 3H] ratio, in basal and stimulation conditions. Present results indicate the existence of changes in PGS synthesis and modulation in VSMC from a conduit-artery of SHR and support the pathophysiological role proposed for matrix proteoglycans in the vascular wall changes associated to hypertension and related vascular diseases as atherosclerosis. 相似文献
4.
Apoptosis is a physiologic form of cell death present in many disease conditions. When the balance of mitosis versus apoptosis is altered, tumor-like growth or degeneration of tissues may ensue. This appears to occur in several diseases, including those of the cardiovascular system, where apoptosis plays a key role in atherosclerosis and restenosis following angioplasty. Since c-myc is upregulated in the pathogenesis of these diseases, we chose to study the sequential morphologic features of programmed cell death in vascular smooth muscle cells induced by c-myc and by the adenovirus early gene E1A. Morphology and timed events in apoptotic cell cultures were analyzed by scanning electron microscopy, transmission electron microscopy, and time-lapse videomicroscopy. We observed that both c-myc-and E1A-induced apoptosis (in serum-free medium) resulted in numerous, tightly packed clusters of apoptotic blebs, as well as in one or two asymmetrically larger blebs. Transmission electron microscopy analysis revealed the larger blebs contained mostly nuclear chromatin, whereas the many smaller fragments often had little or no chromatin. Time-lapse studies showed that apoptosis was induced at a slower rate in cells stably transfected with c-myc versus those stably transfected with E1A. The early changes of apoptosis, including cell shrinkage and intense blebbing, occurred in under 5 min in both cells. Slight alterations such as cell size and further rounding occurred up to 8 h following the initial changes of apoptosis. Rather than being a part of the apoptotic response, release from the culture floor almost entirely resulted from movement of the culture flask. These studies provide a framework of timed morphologic events for future mechanistic investigation into the key aspects of myc-and E1A-induced apoptosis in vascular smooth muscle. 相似文献
5.
James G. Walmsley 《Journal of microscopy》1983,131(3):361-375
A technique of embedding, sectioning and analysis has been developed for studying the orientation and proportional composition of smooth muscle in the straight portions of human major cerebral arteries. Various distortions, which occurred during processing and sectioning, were measured quantitatively. Nerve fibres were implanted as a reference frame within the paraffin block containing an arterial bifurcation. The nuclei of smooth muscle have been treated as three-dimensional vectors of cellular orientation. The projected length in the plane of the section and the section thickness were used to define section pitch. To relate these vectors to the overall geometry of cerebral arteries they were transformed such that the resultant pitch would be the same as that observed if the sections were cut normal to the arterial longitudinal axis. Matrix transformations of nuclear vectors were of the expansion and Eulerian forms. The average pitch for the five sections from three straight portions was ?0.22° ± 2.36 (SD of five means) with a range between ?2.8° and 3.2°. Because this pitch is small it is possible to use 6.9 μm thick longitudinal and mid-plane sections of straight arteries to obtain estimates of proportional composition. Stereological point counting was used to determine that smooth muscle comprised 72.0% ± 4.76 (SD for ten segments) of the tunica media in cerebral arteries. 相似文献
6.
James G. Walmsley 《Journal of microscopy》1983,131(3):377-389
Two approaches were developed for determining reference axes and positions associated with bending tubular arteries and bifurcations from the human middle cerebral branching system. In the region of the bifurcation the orientation of groups of two-dimensional nuclear vectors was determined by the method of roses and average alignment. The reference axes and positions for these groups were determined from physical reconstructions of two bifurcations. The overall pattern in the bifurcation region was found to be random but there were significant differences between group orientations. The three-dimensional arterial direction vectors for three sections, from the bending arteries of one bifurcation, were determined. By taking the dot product of the three-dimensional nuclear vectors and these direction vectors, the pitch of individual nuclei was determined. The average pitch was 2.45° ± 3.20 (SD for N = 8) with the largest mean pitch (6.14°) in the branch with the greatest curvature. Possible use of the three-dimensional approach for determination of smooth muscle orientation at bifurcations is discussed in general terms. 相似文献
7.
8.
Despite considerable research into the pathogenesis of idiopathic headaches, such as migraine, the pathophysiological mechanisms underlying them remain poorly understood. Although it is well established that the trigeminal nerve becomes activated during migraine, the consequences of this activation remain controversial. One theory, based on preclinical observations, is that activation of trigeminal sensory fibers leads to a painful neurogenic inflammation within the meningeal (dural) vasculature mediated by neuropeptide release from trigeminal sensory fibres and characterized by plasma protein extravasation, vasodilation, and mast cell degranulation. Effective antimigraine agents such as ergots, triptans, opioids, and valproate inhibit preclinical neurogenic dural extravasation, suggesting that this activity may be a predictor of potential clinical efficacy of novel agents. However, several clinical trials with other agents that inhibit this process preclinically have failed to show efficacy in the acute treatment of migraine in man. Alternatively, it has been proposed that painful neurogenic vasodilation of meningeal blood vessels could be a key component of the inflammatory process during migraine headache. This view is supported by the observation that jugular plasma levels of the potent vasodilator, calcitonin gene-related peptide (CGRP) are elevated during the headache and normalized by successful sumatriptan treatment. Preclinically, activation of trigeminal sensory fibers evokes a CGRP-mediated neurogenic dural vasodilation, which is blocked by dihydroergotamine, triptans, and opioids but unaffected by NK1 receptor antagonists that failed in clinical trials. These observations suggest that CGRP release with associated neurogenic dural vasodilation may be important in the generation of migraine pain, a theory that would ultimately be tested by the clinical testing of a CGRP receptor antagonist. 相似文献
9.
Adrenomedullin (AM) was originally identified in the extracts of human pheochromocytoma tissue, but this peptide is now known to be synthesized and secreted from many kinds of cells in the body, including vascular smooth muscle cells, endothelial cells, fibroblasts, cardiac myocytes, epithelial cells, and cancer cells. In this review, we summarize AM-secreting and AM gene-expressing cells in addition to the regulation of secretion and gene expression of AM. Although the data are still limited to deduce the general features of AM gene expression, synthesis, and secretion, AM is assumed to be classified into the new class of biologically active peptides, which is mainly expressed and secreted from non-endocrine type cells by the stimulation with inflammation-related substances. It is also interesting that serious physiological conditions such as inflammation or hypoxia potently stimulate AM expression and release, suggesting its unique physiological function distinct from other known biologically active peptides. 相似文献
10.
Nitric oxide (NO) generation by inducible nitric oxide synthase (iNOS) in the vascular smooth muscle cells (VSMC), may play a role in blood vessel tone regulation. Lipopolysaccharide (LPS) induced iNOS activity and subsequent nitrite production by cultured aortic VSMC, from SHR with an established chronic blood pressure elevation (adult SHR) or during the period preceding the development of hypertension (young SHR) and from age-matched normotensive Wistar (W) rats were compared. Angiotensin II (Ang II) effect was also evaluated. Both basal LPS-induced iNOS activity and nitrite accumulation were significantly lower in young SHR VSMC compared to young W rat cells. In contrast, adult hypertensive and normotensive rat cells did not differ in NO generation. Besides, young SHR cells exhibited a significant smaller iNOS activity and nitrites than adult SHR cells. After 24h-incubation with Ang II, both variables were markedly reduced in all groups. The proportional reduction of iNOS activity and nitrites by Ang II was not different between hypertensive and normotensive rat cells, at any age. However, this Ang II inhibitory effect was greater in both adult SHR and W cells than in VSMC from young rats. In conclusion, a reduced LPS-induced iNOS activity and NO generation was observed in VSMC form spontaneously hypertensive rats before the raise of blood pressure, but not in adult hypertensive rat cells. Additionally, an inhibitory effect of angiotensin II on these variables is described. We can speculate that the impairment in vascular smooth muscle NO production precedes the development of hypertension in SHR and may play a pathophysiologic role in the early blood pressure elevation in genetically hypertensive rats. 相似文献
11.
Taenia coli muscle was cooled to 252 K in the presence of the cryoprotectant dimethyl-sulphoxide, at cooling rates known to reduce viability by significantly different amounts. The reduction in viability was known to be related to ice formation. Freeze-substitution and isothermal freeze-fixation studies were carried out to determine the distribution of ice within the muscle at this temperature. Freeze-substitution using ethylene glycol was unsuccessful but a new method, using high concentrations of the cryoprotectant as the substituting solvent, was able to maintain ice configurations at this relatively high substitution temperature. The results of freeze-substitution in dimethylsulphoxide were confirmed by isothermal freeze-fixation when both techniques were conducted under identical cooling conditions. The results indicated that the functional differences produced by cooling muscle at either 0·3 K min?1 or 2 K min?1 were related to the distribution of the ice phase within the tissue. 相似文献
12.
A scanning acoustic microscope operating at 600 MHz was used to observe arterioles in a thin sheet of collagenous connective tissue dissected from the submucosa of the guinea-pig small intestine. The arterioles were clearly defined in images made using transmitted ultrasound, and the acoustic attenuation (α) of the arteriolar wall was estimated to be 120 cm?1. Images made using reflected ultrasound did not show the arterioles clearly. 相似文献
13.
Flávia Karina Delella Sérgio Luis Felisbino 《Microscopy research and technique》2010,73(11):1036-1044
Doxazosin (DOX), an α‐adrenoceptor antagonist, induces the relaxation of smooth muscle cell tonus and reduces the clinical symptoms of benign prostatic hyperplasia (BPH). However, the effects of DOX in the prostate stromal microenvironment are not fully known. In a previous study, we showed that DOX treatment for 30 days increased deposition of collagen fibers in the three rat prostatic lobes. Herein, we investigated the effects of DOX on stromal cell ultrastructure and elastic fiber deposition. Adult Wistar rats were treated with DOX (25 mg/kg/day); and the ventral, dorsal, and anterior prostates were excised at 30 days of treatment. The prostatic lobes were submitted to histochemical and stereological‐morphometric analyze and transmission electron microscopy (TEM). Histochemical staining plus stereological analysis of the elastic fiber system showed that DOX‐treated prostatic lobes presented more elaunin and elastic fibers than controls, mainly in the ventral lobe. Ultrastructural analysis showed that fibroblasts and smooth muscle cells from DOX‐treated prostates presented active synthetic phenotypes, evidenced by enlarged rough endoplasmic reticulum and Golgi apparatus cisterns, and confirmed the observation of thickened elaunin fibers. Our findings suggest that, under α‐adrenergic blockade by DOX, the fibroblasts become more active and smooth muscle cells shift from a predominantly contractile to a more synthetic phenotype. The deposition of collagen and elastic system fibers in the prostatic stroma may counterbalance the absence of smooth muscle tone during α‐blockers treatment. Microsc. Res. Tech. 73:1036–1044, 2010. © 2010 Wiley‐Liss, Inc. 相似文献
14.
It has generally been assumed that tumors do not induce lymphangiogenesis and only very recently animal models have been presented showing tumor-induced lymphangiogenesis. We have grown two types of rat tumor cells, 10AS pancreatic carcinoma and C6 glioma cells, on the chorioallantoic membrane (CAM) of chick and quail embryos. The suspended tumor cells rapidly formed solid tumors which invaded the CAM and were vascularized by CAM vessels. When grown on the CAM of quail embryos intratumoral endothelial cells could be specifically stained with the QH1 antibody. In C6 gliomas the vascular pattern was more regular than in 10AS carcinomas. The vessels often grew radially into the glioma and many of them were invested by smooth muscle alpha-actin-positive periendothelial cells. Lymphatics, which were identified by vascular endothelial growth factor receptor-3 (VEGFR-3) in situ hybridization were absent from C6 gliomas, although a weak expression of the lymphangiogenic growth factor, VEGF-C, could be detected in the C6 cells by Northern blot analysis. In contrast, 10AS cells, which expressed high levels of VEGF-C, induced ingrowth of lymphatics into the tumors, with BrdU-labeling rates of about 9% of lymphatic endothelial cells. Our studies demonstrate the heterogeneity of interactions of tumor cells with blood vessels and lymphatics and show that sufficient quantities and/or quality of lymphangiogenic growth factors are crucial for the induction of lymphatics in tumors. 相似文献
15.
Joseph B. Delcarpio Elizabeth G. Underwood R. L. Moses 《Microscopy research and technique》1989,12(1):24-28
A technique for performing light, scanning, and transverse transmission electron microscopy on cultured cells grown within a single tissue culture flask is described. Permanent light microscopy slides are obtained by removing selected portions of the plastic tissue culture vessel and mounting them on glass slides with an aqueous mounting solution. The images obtained from these slides are superior to viewing through the bottom of the flask with an inverted stage microscope. For scanning electron microscopy, selected areas are also cut from the remainder of the vessel and prepared for viewing. The final portion of the culture container is transferred and attached to a new tissue culture vessel and prepared for transmission electron microscopy using alcohol instead of acetone and propylene oxide during dehydration, infiltration, and embedding. 相似文献
16.
Shabnam Asghari Niari Reza Rahbarghazi Roya Salehi Leila Kazemi Sonia Fathi Karkan Mohammad Karimipour 《Microscopy research and technique》2022,85(4):1433-1443
In recent years with regard to the development of nanotechnology and neural stem cell discovery, the combinatorial therapeutic strategies of neural progenitor cells and appropriate biomaterials have raised the hope for brain regeneration following neurological disorders. This study aimed to explore the proliferation and neurogenic effect of PLGA and PLGA–PEG nanofibers on human SH-SY5Y cells in in vitro condition. Nanofibers of PLGA and PLGA–PEG biomaterials were synthesized and fabricated using electrospinning method. Physicochemical features were examined using HNMR, FT-IR, and water contact angle assays. Ultrastructural morphology, the orientation of nanofibers, cell distribution and attachment were visualized by SEM imaging. Cell survival and proliferation rate were measured. Differentiation capacity was monitored by immunofluorescence staining of Map-2. HNMR, FT-IR assays confirmed the integration of PEG to PLGA backbone. Water contact angel assay showed increasing surface hydrophilicity in PLGA–PEG biomaterial compared to the PLGA substrate. SEM analysis revealed the reduction of PLGA–PEG nanofibers' diameter compared to the PLGA group. Cell attachment was observed in both groups while PLGA–PEG had a superior effect in the promotion of survival rate compared to other groups (p < .05). Compared to the PLGA group, PLGA–PEG increased the number of Ki67+ cells (p < .01). PLGA–PEG biomaterial induced neural maturation by increasing protein Map-2 compared to the PLGA scaffold in a three-dimensional culture system. According to our data, structural modification of PLGA with PEG could enhance orientated differentiation and the dynamic growth of neural cells. 相似文献
17.
18.
T. F. Bendtsen J. R. Nyengaard L. Grimelius† & H. J. G. Gundersen 《Journal of microscopy》2002,207(3):211-224
20.
G. H. Haggis 《Journal of microscopy》1986,143(3):275-282
The performance of a commercial double-propane-jet freezer (Balzers QFD 101) has been assessed, for rapid freezing of fresh tissues in freeze-etch work. Samples of diaphragm muscle and intestinal villi were frozen between copper sheets, with a spacer to give 20–30μm thickness of tissue. Fracture cuts were made with the Balzers BAF 400 freeze-etch microtome within 5–10μm of a freezing face (i.e. a tissue face in contact with the copper sheets of the frozen sandwich). After some modifications to the QFD 101, replicas showing no evidence of ice were obtained of muscle cells, although for intestinal epithelial cells some evidence of ice formation was found. Infiltration with 5% glycerol or dimethylsulphoxide improves the depth of good freezing. Results and problems arising from such infiltration are briefly discussed. 相似文献