Frequent degenerative joint diseases, known as arthritis, are characterized by joint inflammation and cartilage breakdown. Various arthritis types are traditionally managed with intraarticular injections of hyaluronan or its derivatives. However, intravenous administration of hyaluronan is emerging as a vital alternative, particularly because intraarticular injections can be challenging for clinicians when targeting small or swollen joints. Pharmacokinetics of intravenously and intraperitoneally administered middle-Mw hyaluronan were studied in an adjuvant-induced arthritis mouse model alongside therapeutic effects. Using 13C-, biotin- and fluoresce-labeling, we found hyaluronan accumulated in inflamed joint tissues while distribution in other organs remained similar to healthy controls. Repeated administrations significantly reduced arthritis symptoms like swelling and redness, RANKL, inducible nitric oxide synthase, COMP and prostaglandin E2 levels. Moreover, hyaluronan treatment prevented dextran-FITC penetration into inflamed paws suggesting reduced vascular permeability at the site of inflammation. These findings support systemic hyaluronan administration as a promising arthritis treatment strategy.
- MeSH
- artritida experimentální * farmakoterapie patologie metabolismus MeSH
- klouby účinky léků patologie metabolismus MeSH
- kyselina hyaluronová * farmakokinetika aplikace a dávkování farmakologie terapeutické užití MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Cachectic rheumatoid arthritis, the less frequent form of the disease, is associated with loss of fat mass and often more severe course of the disease. Its experimental model represents rat adjuvant arthritis (AA) characterized by edema, lack of appetite, sharp body weight and fat loss. As individual fat depots display functional differences, here we studied lipolytic activity and sensitivity to lipolytic stimuli of nodeless epididymal fat (eWAT) and perinodal mesenteric fat (mWAT) depots at the peak of AA. We also examined changes in catecholamine and cytokine levels involved in lipolysis in plasma and/or isolated adipocytes from both WATs to identify the contribution of local, adipocyte-based processes and/or systemic events to adiposity loss in cachectic rheumatoid arthritis. AA was induced to male Lewis rats by complete Freund's adjuvant. Groups of ad libitum-fed and pair-fed controls were used to distinguish the effects of food restriction from inflammation-induced cachexia. Adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and its phosphorylated form (pHSL) were analyzed by western blot. CRP and catecholamine levels in plasma or adipocyte lysates were determined using ELISA kits. Cytokine-induced neutrophil chemoattractant-1 (CINC-1/CXCL1), monocyte chemoattractant protein-1 (MCP-1/CCL2), IL-1β, IL-6, IL-10 and leptin in adipocyte lysate were analyzed by quantitative protein microarray. Plasma glycerol and FFA were measured spectrophotometrically. AA rats developed severe cachexia, with lower adiposity in mWAT compared to normal and pair-fed controls, whereas in eWAT the adiposity was similarly reduced in AA and pair-fed groups. ATGL levels in both WATs were not affected by AA or pair feeding. AA upregulated levels of HSL, pHSL and pHSL/HSL ratio in mWAT, whereas none of these parameters has changed in eWAT of AA rats or in either WATs of pair-fed rats. In AA rats plasma glycerol was elevated, whereas FFA concentration was reduced. Plasma norepinephrine and epinephrine were increased in AA compared with both groups of controls. In eWAT adipocytes, AA but not pair feeding, upregulated norepinephrine levels. In mWAT adipocytes, AA rats showed higher epinephrine levels than pair-fed controls. Leptin levels in both WATs were depleted in AA animals in accordance with body weight loss. None of the measured cytokines in eWAT and mWAT was enhanced. Our results demonstrate augmented lipolytic activity in mWAT and not eWAT during cachectic arthritis. The adipocyte-derived cytokines do not seem to contribute to activated lipolysis. We first demonstrated enhanced presence of norepinephrine in perinodal adipocytes that may contribute to the regulation of local lipolytic activity by auto/paracrine fashion and thus provide independent fuel supply to activated lymph nodes.
- MeSH
- adrenalin biosyntéza MeSH
- artritida experimentální imunologie metabolismus MeSH
- biologické markery MeSH
- C-reaktivní protein MeSH
- epididymis metabolismus MeSH
- humorální imunita MeSH
- krysa rodu rattus MeSH
- lipolýza MeSH
- mezenterium metabolismus MeSH
- modely nemocí na zvířatech MeSH
- sterolesterasa metabolismus MeSH
- tukové buňky metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVES: Notch ligands and receptors have recently been shown to be differentially expressed in osteoarthritis (OA). We aim to further elucidate the functional role of Notch signalling in OA using Notch1 antisense transgenic (Notch1 AS) mice. METHODS: Notch and hedgehog signalling were analysed by real-time PCR and immunohistochemistry. Notch-1 AS mice were employed as a model of impaired Notch signalling in vivo. Experimental OA was induced by destabilisation of the medial meniscus (DMM). The extent of cartilage destruction and osteophyte formation was analysed by safranin-O staining with subsequent assessment of the Osteoarthritis Research Society International (OARSI) and Mankin scores and µCT scanning. Collagen X staining was used as a marker of chondrocyte hypertrophy. The role of hairy/enhancer of split 1 (Hes-1) was investigated with knockdown and overexpression experiments. RESULTS: Notch signalling was activated in human and murine OA with increased expression of Jagged1, Notch-1, accumulation of the Notch intracellular domain 1 and increased transcription of Hes-1. Notch1 AS mice showed exacerbated OA with increases in OARSI scores, osteophyte formation, increased subchondral bone plate density, collagen X and osteocalcin expression and elevated levels of Epas1 and ADAM-TS5 mRNA. Inhibition of the Notch pathway induced activation of hedgehog signalling with induction of Gli-1 and Gli-2 and increased transcription of hedgehog target genes. The regulatory effects of Notch signalling on Gli-expression were mimicked by Hes-1. CONCLUSIONS: Inhibition of Notch signalling activates hedgehog signalling, enhances chondrocyte hypertrophy and exacerbates experimental OA including osteophyte formation. These data suggest that the activation of the Notch pathway may limit aberrant hedgehog signalling in OA.
- MeSH
- artritida experimentální metabolismus MeSH
- chondrocyty metabolismus MeSH
- kloubní chrupavka metabolismus MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- membránové glykoproteiny metabolismus MeSH
- myši transgenní MeSH
- myši MeSH
- osteoartróza metabolismus MeSH
- osteofyt metabolismus MeSH
- receptor Notch1 metabolismus MeSH
- signální transdukce MeSH
- stupeň závažnosti nemoci MeSH
- transkripční faktor HES1 metabolismus MeSH
- transportní proteiny metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Carnosine (CARN) is an anti-glycating agent able to quench superoxide, and to neutralize 4-hydroxynonenal. Trolox-carnosine (CARN-T) was synthesized because of its resistance against degradation and to improve CARN antioxidant capacity. We evaluated the impact of trolox (TRO), CARN and its derivative CARN-T on oxidative stress (OS) in brain during rat adjuvant arthritis (AA). The experiments were done on healthy, control arthritic and arthritic animals with administration of CARN 150 mg/kg b.w., TRO 41 mg/kg b.w. and CARN-T 75 mg/kg b.w. in a daily dose during 28 days. Antioxidants did not affect the body weight on day 14, but on day 28 TRO enhanced the weight reduction. On day 14 and 28 CARN-T and TRO reduced arthritic score. IL-1beta, MCP-1 and MMP-9 were measured in plasma on day 14. MCP-1 was decreased by CARN-T and TRO. All antioxidants reduced IL-1beta and MMP-9 levels. Malondialdehyde, 4-hydroxynonenal and protein carbonyls were increased in brain. CARN, CARN-T and TRO prevented higher lipid and protein oxidation in brain. CARN and CARN-T caused no weight reduction like TRO that has an advantage in inflammatory arthritis. Moreover the antioxidants administered had a similar therapeutic effects on arthritic score, markers of inflammation in plasma and OS in brain.
- MeSH
- antioxidancia farmakologie terapeutické užití MeSH
- artritida experimentální farmakoterapie metabolismus MeSH
- chromany (dihydrobenzopyrany) farmakologie terapeutické užití MeSH
- karnosin analogy a deriváty farmakologie terapeutické užití MeSH
- krysa rodu rattus MeSH
- mozek účinky léků metabolismus MeSH
- oxidační stres účinky léků fyziologie MeSH
- potkani inbrední LEW MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We studied anxiety-like behavior in the elevated plus-maze (EPM) tests in male Lewis rats on days 2 and 4 of adjuvant arthritis (AA). In plasma we analyzed C-reactive protein (CRP), albumin, ACTH, corticosterone, in the hippocampus the mRNA expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), corticotrophin releasing factor (CRH), NADPH oxidases NOX1 and NOX2, and inducible NO-synthase (iNOS). EPM tests showed a higher anxiety index in AA rats on days 2 and 4 and reduction of total entries. On days 2 and 4 we found reduced plasma albumin, enhanced CRP, ACTH and corticosterone, and in the hippocampus enhanced mRNA for NOX1 and IL-1β in AA rats, on day 4 we found enhanced mRNAs for iNOS and IL-6, and reduced mRNA for CRH. The mRNA for NOX2 did not change on any experimental day. These results suggest enhanced anxiety, as well as locomotor impairment during the early phase of AA that correlate with enhanced mRNA expressions of parameters of oxidative stress NOX1, iNOS, and inflammatory cytokines IL-1β and IL-6 in the hippocampus.
- MeSH
- artritida experimentální komplikace metabolismus psychologie MeSH
- bludiště - učení fyziologie MeSH
- hipokampus metabolismus MeSH
- interleukin-1beta biosyntéza MeSH
- interleukin-6 biosyntéza MeSH
- krysa rodu rattus MeSH
- messenger RNA biosyntéza MeSH
- NADH, NADPH oxidoreduktasy biosyntéza MeSH
- potkani inbrední LEW MeSH
- synthasa oxidu dusnatého, typ II biosyntéza MeSH
- úzkost komplikace metabolismus psychologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
OBJECTIVE: Rheumatoid arthritis (RA) is associated with enhanced pro-inflammatory cytokine levels, pain, anorexia, and cognitive changes. The enhanced production of cytokines appears before the full manifestation of the disease. So far, any experimental data on behavioral effects of early arthritis are lacking. In the present series we describe anorexia early changes in, pain hyper-sensitivity and altered cognitive behavior during the first four days of adjuvant arthritis in rats (AA), when no clinical signs are yet apparent. METHODS: AA was induced to male Lewis rats by a single injection of complete Freund's adjuvant (cFA) at the base of the tail. Plasma leptin and ghrelin were measured using specific RIA methods. Gene expressions for food-regulatory peptides, neuropeptide-Y (NPY) and interleukin-1β (IL-1β) in the hypothalamic arcuate nuclei (nARC), were quantitated by TaqMan real-time PCR. Pain sensation was measured on all four limbs and tail by the plantar test. Cognitive functions were tested in the Morris water maze (MWM). RESULTS: Levels of orexigenic ghrelin as well as mRNA expression of orexigenic NPY in nucleus arcuatus (nRC)re significantly enhanced on day 2 of AA only. Reduced body weight and food intake persisted by day 4 with the most profound reduction on day 2. The mRNA for anorexigenic IL-1β in the nARC was significantly enhanced on days 2 and 4. Enhanced pain sensitivity was observed on day 2, as was the cognitive impairment given by longer time to find the hidden platform, longer time spent in thigmotaxis zone, and longer trajectory. The less effective strategy used to find the hidden platform was observed up to the day 4 of AA. CONCLUSIONS: Early stage of AA brings about reduced body weight, food intake, and activation of central orexigenic pathways. The observed anorexia could be ascribed to the over-expression of anorexigenic IL-1β which dominates over the NPY orexigenic effects. On day 2 of AA higher pain sensitivity and cognitive impairment appear. All the observed change tend to recover by day 4 of the disease.
- MeSH
- artritida experimentální komplikace metabolismus MeSH
- časové faktory MeSH
- exprese genu MeSH
- ghrelin krev MeSH
- hyperalgezie etiologie MeSH
- interleukin-1beta genetika MeSH
- kognitivní poruchy etiologie MeSH
- krysa rodu rattus MeSH
- leptin krev MeSH
- messenger RNA analýza MeSH
- nechutenství etiologie MeSH
- neuropeptid Y genetika MeSH
- nucleus arcuatus hypothalami chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
11beta-Hydroxysteroid dehydrogenase 1 (11HSD1) regulates local glucocorticoid activity and plays an important role in various diseases. Here, we studied whether arthritis modulates 11HSD1, what is the role of pro-inflammatory cytokines in this process and whether altered local metabolism of glucocorticoids may contribute to the feedback regulation of inflammation. Adjuvant arthritis increased synovial 11HSD1 mRNA and 11-reductase activity but treatments with tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta) antagonists etanercept and anakinra reduced 11HSD1 upregulation. Treatment with carbenoxolone, an 11HSD inhibitor, increased expression of TNF-alpha, cyclooxygenase 2, and osteopontin mRNA without any changes in the plasma levels of corticosterone. Similar changes were observed when arthritic rats were treated with RU486, an antagonist of GR. This study suggests that arthritis upregulates synovial 11HSD1, this upregulation is controlled by TNF-alpha and IL-1beta and that the increased supply of local corticosterone might contribute to feedback regulation of inflammation.
- MeSH
- 11-beta-hydroxysteroiddehydrogenasy genetika metabolismus MeSH
- antagonisté hormonů farmakologie MeSH
- artritida experimentální genetika metabolismus MeSH
- cytokiny metabolismus MeSH
- glukokortikoidy metabolismus MeSH
- izoenzymy genetika metabolismus MeSH
- karbenoxolon farmakologie MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- lidé MeSH
- messenger RNA metabolismus MeSH
- mifepriston farmakologie MeSH
- potkani inbrední LEW MeSH
- protivředové látky farmakologie MeSH
- synoviální membrána cytologie účinky léků metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Parvalbumin (PV) is a calcium-binding protein that is expressed by numerous neuronal subpopulations in the central nervous system. Staining for PV was often used in neuroanatomical studies in the past. Recently, several studies have suggested that PV acts in neurons as a mobile endogenous calcium buffer that affects temporo-spatial characteristics of calcium transients and is involved in modulation of synaptic transmission. In our experiments, expression of PV in the lumbar dorsal horn spinal cord was evaluated using densitometric analysis of immunohistological sections and Western-blot techniques in control and arthritic rats. There was a significant reduction of PV immunoreactivity in the superficial dorsal horn region ipsilateral to the arthritis after induction of the peripheral inflammation. The ipsilateral area and intensity of PV staining in this area were reduced to 38 % and 37 %, respectively, out of the total PV staining on both sides. It is suggested that this reduction may reflect decreased expression of PV in GABAergic inhibitory neurons. Reduction of PV concentration in the presynaptic GABAergic terminals could lead to potentiation of inhibitory transmission in the spinal cord. Our results suggest that changes in expression of calcium-binding proteins in spinal cord dorsal horn neurons may modulate nociceptive transmission.
- MeSH
- artritida experimentální chemicky indukované metabolismus MeSH
- buňky zadních rohů míšních chemie MeSH
- down regulace MeSH
- financování organizované MeSH
- kaolin MeSH
- karagenan MeSH
- krysa rodu rattus MeSH
- lumbosakrální krajina MeSH
- parvalbuminy analýza MeSH
- potkani Wistar MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
Adjuvant arthritis (AA) is a model of chronic inflammation induced by Mycobacterium butyricum and characterized by similar pathophysiological and pathobiochemical changes as rheumatoid arthritis (RA) in humans. In this study the antirheumatic activity of coenzyme Q(10) supplementation was tested not only as to its capability to suppress the inflammation edema of the hind paw and to improve the body weight of the arthritic animals, but also to improve so important biochemical parameters as markers of inflammation and oxidative stress, and of mitochondrial bioenergetics. Despite the unfavorable effects on the rheumatic processes observed by monitoring biometric parameters (hind paw volume, relative body weight, relative weight of spleen), a significant protective effect was observed on the level of mitochondrial energetic and antioxidant disbalance. This finding speaks in favor of CoQ(10) supplementation in rheumatic patients, presumably as combinatory therapy with classical antirheumatics, e.g. NSAIDs.
- MeSH
- artritida experimentální imunologie metabolismus patofyziologie MeSH
- cirkadiánní rytmus imunologie MeSH
- finanční podpora výzkumu jako téma MeSH
- interleukin-1 genetika MeSH
- interleukin-6 genetika MeSH
- krysa rodu rattus MeSH
- neuroimunomodulace fyziologie MeSH
- pro-opiomelanokortin genetika MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
