The aim of the present study was to investigate whether enzyme chondroitinase ABC (ChABC) treatment influences the phenotype of neural progenitor cells (NPCs) derived from injured rat spinal cord. Adult as well as fetal spinal cords contain a pool of endogenous neural progenitors cells, which play a key role in the neuroregenerative processes following spinal cord injury (SCI) and hold particular promise for therapeutic approaches in CNS injury or neurodegenerative disorders. In our study we used in vitro model to demonstrate the differentiation potential of NPCs isolated from adult rat spinal cord after SCI, treated with ChABC. The intrathecal delivery of ChABC (10 U/ml) was performed at day 1 and 2 after SCI. The present findings indicate that the impact of SCI resulted in a decrease of all NPCs phenotypes and the ChABC treatment, on the contrary, caused an opposite effect.
- MeSH
- chondroitinasa ABC farmakologie terapeutické užití MeSH
- fenotyp MeSH
- kmenové buňky enzymologie účinky léků MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- mícha cytologie enzymologie účinky léků MeSH
- neurony enzymologie účinky léků MeSH
- poranění míchy enzymologie farmakoterapie patologie MeSH
- potkani Wistar MeSH
- výsledek terapie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Knowledge on the involvement of spinal COX-1 and COX-2 in pain due to osteoarthritis could be useful for better understanding of its pathogenesis and therapy. In this study we have investigated a long-term pattern of expression and production of spinal COX-1 and COX-2 in the model of osteoarthritis induced in rats by injection of monoiodoacetate (MIA) into the knee joint. MIA injection produced thermal hyperalgesia (assessed by the plantar test) and tactile allodynia (measured with von Frey hairs). The pain measures reached maximum on the fifht day, then remained relatively stable. The expression of spinal COX-2 mRNA reached maximum on day 5 (5.2 times; P<0.001) and remained increased until day 31 (4.9 times; P<0.001). Expression of spinal COX-1 mRNA increased gradually reaching maximum on the day 31 (4.5 times; P<0.001) when the relative expression of both genes was almost equal. The production of both proteins was almost similar at the beginning of the experiment. The highest production of COX-2 protein was observed on day 5 after the induction of osteoarthritis (increased 3.9 times). The levels of COX-1 protein increased gradually with maximum on day 31 (3.4 times). The present findings indicate that not only expression of COX-2 mRNA but also that of COX-1 mRNA is significantly increased in the spine during osteoarthritis pain. Thus, in contrast to inflammatory pain, the upregulation of spinal COX-1 may be important in osteoarthritis pain.
- MeSH
- artróza kolenních kloubů enzymologie genetika chemicky indukované MeSH
- bolest epidemiologie genetika chemicky indukované MeSH
- časové faktory MeSH
- cyklooxygenasa 1 biosyntéza genetika MeSH
- cyklooxygenasa 2 biosyntéza genetika MeSH
- enzymová indukce MeSH
- financování organizované MeSH
- hyperalgezie enzymologie genetika chemicky indukované MeSH
- krysa rodu rattus MeSH
- kyselina jodoctová MeSH
- membránové proteiny biosyntéza genetika MeSH
- měření bolesti MeSH
- messenger RNA MeSH
- mícha enzymologie MeSH
- modely nemocí na zvířatech MeSH
- potkani Wistar MeSH
- práh bolesti MeSH
- reakční čas MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
BACKGROUND: Prostate specific membrane antigen (PSMA), also called glutamate carboxypeptidase II (GCPII), is a target enzyme for diagnosis and treatment of prostate cancer. Moreover, it is upregulated in the vasculature of most solid tumors and is therefore a potential target for the generation of novel antineoplastics. In this context, we analyze the possibility of using rat and pig as animal models for enzymologic and in vivo studies. METHODS: We prepared the recombinant extracellular part of human, rat, and pig GCPII in S2 cell media and characterized the activity and inhibition profiles of the three orthologs by radioenzymatic assay. We performed Western blot analysis of GCPII expression in human, rat, and pig tissues using the monoclonal antibody GCP-04 and confirmed these findings by activity measurements and immunohistochemistry. RESULTS: The three recombinant proteins show similar specific enzymatic activities and inhibition profiles. Tissue expression analysis revealed that most of the pig and human tissues show at least some GCPII-positivity, while the expression pattern in rat is more restricted. Moreover, tissues such as prostate and testes exhibit different GCPII expression levels among the species studied. CONCLUSIONS: The rat and pig orthologs of GCPII seem to be suitable to approximate human GCPII in enzymologic studies. However, the diffuse expression pattern of GCPII in animal and human tissues could be a caveat for the potential utilization of GCPII-targeted anticancer drugs. Furthermore, variations in GCPII tissue distribution among the species studied should be considered when using rat or pig as models for antineoplastic drug discovery.
- MeSH
- druhová specificita MeSH
- financování organizované MeSH
- krysa rodu rattus MeSH
- ledviny enzymologie patologie MeSH
- lidé MeSH
- mícha enzymologie patologie MeSH
- miniaturní prasata MeSH
- modely u zvířat MeSH
- molekulární sekvence - údaje MeSH
- potkani inbrední LEW MeSH
- prasata MeSH
- prostata enzymologie patologie MeSH
- prostatický specifický antigen analýza genetika metabolismus MeSH
- regulace genové exprese enzymů MeSH
- sekvence aminokyselin MeSH
- testis enzymologie patologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
Levels of cyclooxygenase-2 (COX-2) mRNA, but not those of COX-1, were reported to be raised significantly after peripheral inflammation in the rat spinal cord. The aim of the present study was to ascertain whether this pattern of COX-2 and COX-1 expression applies also to other pain conditions induced by surgical procedure. Experiments were performed on two types of pain models. In a model of postoperative pain, 1 cm longitudinal incision was made through skin, fascia and muscle of the plantar aspect of the right hind paw in anaesthetized rats. In the second model, peripheral inflammation was induced by unilateral, intraplantar injection of carrageenan in the right hind paw. Carrageenan injection or skin incision produced marked and significant reduction of paw withdrawal latencies to noxious radiant heat stimuli after 2 and 6 hr. Under the acute inflammation 2 and 6 hr after carrageenan injection levels of COX-2 mRNA were markedly raised (7.8 and 15.5 times; P<0.001, respectively) while spinal levels of COX-1 mRNA were not significantly altered (n.s.). In contrast, spinal levels of COX-2 mRNA were raised less markedly in a model of postoperative pain (4.9 times at 2 hr; P<0.001 and 2.9 times (n.s.) at 6 hr after surgery) whilst levels of COX-1 mRNA in the lumbar spine were increased significantly (2.3 times; P<0.001) 6 hr after surgery. The present findings indicate that expression of COX-2 mRNA in the spine is less dominant in postoperative pain than in inflammatory pain and that spinal COX-1 mRNA is upregulated in postoperative pain.
- MeSH
- cyklooxygenasa 1 genetika MeSH
- cyklooxygenasa 2 genetika MeSH
- exprese genu genetika MeSH
- financování organizované MeSH
- karagenan aplikace a dávkování toxicita MeSH
- krysa rodu rattus MeSH
- měření bolesti metody MeSH
- messenger RNA genetika metabolismus MeSH
- mícha enzymologie metabolismus MeSH
- modely nemocí na zvířatech MeSH
- polymerázová řetězová reakce s reverzní transkripcí metody MeSH
- pooperační bolest enzymologie genetika patofyziologie MeSH
- potkani Wistar MeSH
- upregulace genetika MeSH
- zadní končetina enzymologie chirurgie metabolismus MeSH
- zánět enzymologie genetika chemicky indukované MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
Zápalový proces je nevyhnutnou odpoveďou na poškodenie rôzneho pôvodu, ktorý sa odohráva aj v centrálnom nervovom systéme (CNS). Hoci hromadiace sa údaje presvedčivo dokumentujú, že chronický zápalový proces je dôležitou súčasťou neurodegenratívnych chorôb, akými sú Alzheimerova choroba, Parkinsonova choroba, Huntingtonova choroba, menej je známe o akútnom zápalovom procese v CNS. Zatiaľ čo neurodegeneratívne choroby sú charakterizované neprítomnosťou významnej infiltrácie z krvi pochádzajúcich mononukleárnych buniek, prítomnosť mikroglie/makrofágov je typická pre akútnu neuroinflamáciu. V priebehu fokálnej, netraumatickej neuroinflamácie, ktorá je vyvolaná mikroinjekciou zymosanu do parenchýmu miechy, dochádza k miestnej aktivizácii mikroglie/makrofágov. V našej práci sme zisťovali vznik, rozsah a vývoj fokálnej lézie a priebeh aktivizácie mikroglie/makrofágov po stereotaxickej aplikácii zymosanu (100 nl) do bočných povrazcov miechy v akútnom období po podaní zymosanu (6 hod.–8 dní). Intenzívnu aktivitu NADPH-diaforázy v makrofágoch lézie sme sledovali 1–4 dni po zymosanovej injekcii, v ďalsích dňoch prežívania klesala. Naše výsledky potvrdzujú aktívnu úlohu mikroglie/makrofágov pri fokálnej netraumatickej neuroinflamácii a podporujú inovatívne názory na procesy akútnej neuroinflamácie v CNS.
Inflammation is an inevitable response on injury of different origin that takes place, as well, in the central nervous system (CNS). Although accumulating evidence suggests that chronic inflammation plays an important part in neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, less is known about the processes of the acute CNS inflammation. While neurodegenerative diseases are characterized by the lack of the prominent infiltrates of blood-derived mononuclear cells the involvement of microglia/macrophages is typical for the acute neuroinflammation. Focal non-traumatic neuroinflammation produced by microinjection of zymosan to the parenchyma of spinal cord induces local activation of microglia/macrophages. In our study, we evaluated focal lesion progress and microglia/macrophages response following stereotaxic application of zymosan (100 nl) to the lateral funiculi of the spinal cord in acute post-injection time period (6 h–8 days). Intensive expression of NADPH-diaphorase in lesion macrophages was observed 1–4 days after zymosan injection, in the next days its activity declined. Our results confirm active role of microglia/macrophages in focal non-traumatic neuroinflammation and strengthen innovative view on processes of acute inflammation in CNS.
- MeSH
- finanční podpora výzkumu jako téma MeSH
- krysa rodu rattus MeSH
- makrofágy enzymologie MeSH
- mícha enzymologie patologie MeSH
- mikroglie imunologie MeSH
- modely u zvířat MeSH
- NADPH-dehydrogenasa analýza MeSH
- neurogenní zánět imunologie patologie MeSH
- zvířata MeSH
- zymosan aplikace a dávkování MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- aferentní nervové dráhy enzymologie fyziologie MeSH
- axony enzymologie MeSH
- finanční podpora výzkumu jako téma MeSH
- imunohistochemie metody MeSH
- lidé MeSH
- mícha cytologie enzymologie MeSH
- motorické neurony enzymologie MeSH
- NADPH-dehydrogenasa metabolismus MeSH
- nervová vlákna enzymologie MeSH
- nervus ischiadicus metabolismus MeSH
- počet buněk metody MeSH
- proteiny nervové tkáně metabolismus MeSH
- psi MeSH
- rizotomie metody MeSH
- spinální ganglia cytologie MeSH
- stilbamidiny metabolismus MeSH
- synthasa oxidu dusnatého metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
