Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
Nestr.
Idiopatický pes equinovarus (angl. clubfoot) je vrozená vada chodidla a bérce. Ačkoli se jedná o nejčastější vrozenou vadu pohybového aparátu, stále není jasná přesná etiopatogeneze tohoto onemocnění. Naše předchozí výsledky ukazují, že patologická tkáň na mediální straně nohy je složena převážně z fibrózních a pro-fibrózních proteinů. V tomto projektu bude aplikováno několik látek, které mají schopnost tuto fibrózní tkáň rozvolnit nebo látky, které zabraňují dalšímu sesíťování fibrózních proteinů za účelem uvolnění napětí v kontrahované tkáni nohy. Aplikace testovaných látek bude probíhat jednak přímo na vyoperovaných tkáních z kontrahované mediální strany postižené nohy a dále pak na buněčných kulturách izolovaných z těchto tkání. Poté budou u vzorků zjištěny změny v morfologii a složení mezibuněčné hmoty a viabilitě buněk a budou charakterizovány jejich mechanické vlastnosti. Výsledky budou statisticky zhodnoceny. Vybrané látky mohou být v budoucnu využity pro terapeutické působení na fibrotickou tkáň a pro vylepšení konzervativní terapie onemocnění pes equinovarus.; Idiopathic pes equinovarus (clubfoot) is a congenital deformity of the lower leg. It is the most common birth defects of the musculoskeletal system; however, the specific etiology of this disease remains unclear. Our previous results show that rigid tissue on the deformed foot is composed mainly of fibrotic and pro-fibrotic proteins. This project suggests application of several compounds which are either able to cleave the fibrous proteins or to prevent further crosslinking of the fibrous tissue in order to release the tension of foot contraction. The compounds will be applied either directly on tissue samples dissected from the contracted side of clubfoot or they will be added into the medium of cell cultures isolated from the contracted tissue. Impact of the treatments on the tissue samples and cell cultures will be analyzed comparing the changes in the extracellular matrix morphology and composition and in cell viability. Biomechanical properties of the treated samples will be also investigated by tensile tests. Selected compounds can improve conservative therapy of the clubfoot.
- MeSH
- antifibrotické látky terapeutické užití MeSH
- enzymoterapie metody MeSH
- fibróza farmakoterapie MeSH
- kolagenasy terapeutické užití MeSH
- lidé MeSH
- pes equinovarus etiologie farmakoterapie patofyziologie MeSH
- separace buněk MeSH
- Check Tag
- lidé MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- farmakoterapie
- podiatrie
- genetika, lékařská genetika
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
Idiopathic pes equinovarus (clubfoot) is a congenital deformity of the feet and lower legs. Clubfoot belongs to a group of fibro-proliferative disorders but its origin remains unknown. Our study aimed to achieve the first complex proteomic comparison of clubfoot contracted tissue of the foot (medial side; n = 16), with non-contracted tissue (lateral side; n = 13). We used label-free mass spectrometry quantification and immunohistochemistry. Seven proteins were observed to be significantly upregulated in the medial side (asporin, collagen type III, V, and VI, versican, tenascin-C, and transforming growth factor beta induced protein) and four in the lateral side (collagen types XII and XIV, fibromodulin, and cartilage intermediate layer protein 2) of the clubfoot. Comparison of control samples from cadavers brought only two different protein concentrations (collagen types I and VI). We also revealed pathological calcification and intracellular positivity of transforming growth factor beta only in the contracted tissue of clubfoot. Most of the 11 differently expressed proteins are strongly related to the extracellular matrix architecture and we assume that they may play specific roles in the pathogenesis of this deformity. These proteins seem to be promising targets for future investigations and treatment of this disease. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.
- MeSH
- dítě MeSH
- extracelulární matrix - proteiny metabolismus MeSH
- hmotnostní spektrometrie MeSH
- kalcinóza MeSH
- lidé MeSH
- pes equinovarus etiologie metabolismus MeSH
- předškolní dítě MeSH
- prospektivní studie MeSH
- proteom MeSH
- transformující růstový faktor beta metabolismus MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
The heart is characterized by a remarkable degree of heterogeneity. Since different cardiac pathologies affect different cardiac regions, it is important to understand molecular mechanisms by which these parts respond to pathological stimuli. In addition to already described left ventricular (LV)/right ventricular (RV) and transmural differences, possible baso-apical heterogeneity has to be taken into consideration. The aim of our study has been, therefore, to compare proteomes in the apical and basal parts of the rat RV and LV. Two-dimensional electrophoresis was used for the proteomic analysis. The major result of this study has revealed for the first time significant baso-apical differences in concentration of several proteins, both in the LV and RV. As far as the LV is concerned, five proteins had higher concentration in the apical compared to basal part of the ventricle. Three of them are mitochondrial and belong to the "metabolism and energy pathways" (myofibrillar creatine kinase M-type, L-lactate dehydrogenase, dihydrolipoamide dehydrogenase). Myosin light chain 3 is a contractile protein and HSP60 belongs to heat shock proteins. In the RV, higher concentration in the apical part was observed in two mitochondrial proteins (creatine kinase S-type and proton pumping NADH:ubiquinone oxidoreductase). The described changes were more pronounced in the LV, which is subjected to higher workload. However, in both chambers was the concentration of proteins markedly higher in the apical than that in basal part, which corresponds to the higher energetic demand and contractile activity of these segments of both ventricles.
- MeSH
- 2D gelová elektroforéza MeSH
- chaperon hsp60 metabolismus MeSH
- chromatografie kapalinová MeSH
- dihydrolipoamiddehydrogenasa metabolismus MeSH
- energetický metabolismus MeSH
- kreatinkinasa, forma MM metabolismus MeSH
- L-laktátdehydrogenasa metabolismus MeSH
- lehké řetězce myosinu metabolismus MeSH
- mitochondriální proteiny metabolismus MeSH
- potkani Wistar MeSH
- proteomika * MeSH
- respirační komplex I metabolismus MeSH
- srdeční komory enzymologie metabolismus MeSH
- svalové proteiny izolace a purifikace metabolismus MeSH
- tandemová hmotnostní spektrometrie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
INTRODUCTION: The unique pulp-dentin complex demonstrates strong regenerative potential, which enables it to respond to disease and traumatic injury. Identifying the proteins of the pulp-dentin complex is crucial to understanding the mechanisms of regeneration, tissue calcification, defense processes, and the reparation of dentin by dental pulp. The lack of knowledge of these proteins limits the development of more efficient therapies. METHODS: The proteomic profile of human tooth pulp was investigated and compared with the proteome of human dentin and blood. The samples of tooth pulp were obtained from 5 sound permanent human third molars of 5 adults (n = 5). The extracted proteins were separated by 2-dimensional gel electrophoresis, analyzed by nano-liquid chromatography tandem mass spectrometry, and identified by correlating mass spectra to the proteomic databases. RESULTS: A total of 342 proteins were identified with high confidence, and 2 proteins were detected for the first time in an actual human sample. The identified tooth pulp proteins have a variety of functions: structural, catalytic, transporter, protease activity, immune response, and many others. In a comparison with dentin and blood plasma, 140 (pulp/dentin) shared proteins were identified, 37 of which were not observed in plasma. It can be suggested that they might participate in the unique pulp-dentin complex. CONCLUSIONS: This proteomic investigation of human tooth pulp, together with the previously published study of human dentin, is one of the most comprehensive proteome lists of human teeth to date.
- MeSH
- 2D gelová elektroforéza MeSH
- chromatografie kapalinová MeSH
- dentin chemie MeSH
- dospělí MeSH
- energetický metabolismus fyziologie MeSH
- hmotnostní spektrometrie MeSH
- imunoproteiny analýza MeSH
- krevní proteiny analýza MeSH
- lidé MeSH
- mezibuněčná komunikace fyziologie MeSH
- mladý dospělý MeSH
- nanotechnologie MeSH
- proliferace buněk MeSH
- proteiny metabolismus MeSH
- proteom analýza klasifikace MeSH
- signální transdukce fyziologie MeSH
- tandemová hmotnostní spektrometrie MeSH
- zubní dřeň chemie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
