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Ultrastructure and histochemistry of rat myocardial capillary endothelial cells in response to diabetes and hypertension

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Cell Research 2005年第7期15卷 532-538页

作者： Ludmila OKRUHLICOVA,Narcis TRIBULOVA1,peter weismann,Ruzena SOTNIKOVA Institute for Heart Research Slovak Academy of Sciences 840 05 Bratislava Slovak Republic Institute for Heart Research Slovak Academy of Sciences 840 05 Bratislava Slovak Republic Medical Faculty Comenius University Bratislava Slovak Republic Institute of Experimental Pharmacology Slovak Academy of Sciences Bratislava Slovak Republic

Insufficient growth and rarefaction of capillaries, followed by endothelial dysfunction may represent one of the mostcritical mechanisms involved in heart damage. In this study we examined histochemical and ultrastructural changes inmyocardial capillary endothelium in two models of heart failure streptozotocin-induced diabetes mellitus (STZ) and NO-deficient hypertension in male Wistar rats. Diabetes was induced by a single i.v. dose of STZ (45 mg/kg) and chronic 9-week stage was analysed. To induce NO-deficient hypertension, animals were treated with inhibitor of NO synthase L-nitroarginine methylester (L-NAME) (40 mg/kg) for 4 weeks. Left ventricular tissue was processed for enzyme cata-lytic histochemistry of capillary alkaline phosphatase (AlPh), dipeptidyl peptidase IV (DPP IV), and endothelial NOsynthase/NADPH-diaphorase (NOS) and for ultrastructural analysis. In diabetic and hypertensive rats, lower/absentAlPh and DPP IV activities were found in focal micro-areas. NOS activity was significantly reduced and persisted onlylocally. Quantitative evaluation demonstrated reduction of reaction product intensity of AlPh, DPP and NOS by 49.50%,74.36%, 20.05% in diabetic and 62.93%, 82.71%, 37.65% in hypertensive rats. Subcellular alterations of endothelialcells were found in heart of both groups suggesting injury of capillary function as well as compensatory *** injury was more significant in diabetic animals, in contrast the adaptation was more evident in hypertensiveones. Concluding: both STZ-induced diabetes- and NO-deficient hypertension-related cardiomyopathy were accompa-nied by similar features of structural remodelling of cardiac capillary network manifested as angiogenesis and *** latter was however, predominant and may accelerate disappearance of capillary endothelium contributing to myo-cardial dysfunction.

关键词： rat heart diabetes NO-deficiency capillary ultrastructure histochemistry.

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Autoantibodies against G-Protein-Coupled Receptors Modulate Heart Mast Cells

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Cellular & Molecular Immunology 2007年第2期4卷 127-133页

作者： Ludmila Okruhlicova Rosemarie Morwinski Wolfgang Schulze Sabine Bartel peter weismann Narcisa Tribulova Gerd Wallukat Institute for Heart Research Slovak Academy of Sciences Dubravska cesta 9 PO Box 45 840 05 Bratislava 45 Slovak Republic Max-Delbruck-Center for Molecular Medicine Robert Rssle Str. 10 D-13125 Berlin Germany Max-Delbruck-Center for Molecular Medicine Robert Rssle Str. 10 D-13125 Berlin Germany Medical Faculty Comenius University Institute of Anatomy Sasinkova 2 813 72 Bratislava Slovak Republic Institute for Heart Research Slovak Academy of Sciences Dubravska cesta 9 PO Box 45 840 05 Bratislava 45 Slovak Republic

Mast cells are believed to be involved in myocardial tissue remodelling under pathophysiological conditions. We examined the effects of autoantibodies against G-protein-coupled receptors in sera of patients with heart diseases on myocardial mast cells in the cultured neonatal Sprague-Dawley rat heart cells. Cells collected at day 3 and 10 of the culture were preincubated with autoantibodies against α-adrenoceptor and angiotensin Ⅱ ATl-receptor, agonist phenylephrine and angiotensin Ⅱ, and control IgG. The pretreated cultured cells were stained for selected mast cell markers tryptase, chymase and TNF-α The cultured cells were also processed for observation with electron microscopy. The autoantibodies-treatment of the 3-day cultured cells caused both increased intensity of immunofluorescence （p 〈 0.05） and their enlarged diameters of the mast cells when compared to age-matched ones. In contrast, the fluorescence of preincubated 10-day-old mast cells was decreased compared with controls （p 〈 0.01）. In control samples, the fluorescence of 10-day-old mast cells was significantly higher than that of 3-day-old ones （p 〈 0.001）. Results of electron microscopy examination demonstrated there was an increased granulation of treated 3-day-old mast cells, while a degranulation of mast cells at day 10 of application. The results suggest the modulation effect of the autoantibodies against G-protein-coupled receptors on mast cells, indicating a potential functional link between the autoantibodies against G-protein-coupled receptors and the mast cells in progression of heart disease.

关键词： mast cell autoantibody rat heart cell culture immunofluorescence

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