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  The Effect of Ceftriaxone on the Expression of Glutamate transporter GLT -1 in the Spinal cord and on the Extracellular Concentration of Glutamate in the Cerebrospinal Fluid in the Chronic Constriction Injury Rat model of Neuropathic Pain.
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 Ophav:
Merry, Ann1, Forfatter
Bjerrum, Ole J.1, Vejleder
Tilknytninger:
1Det Sundhedsvidenskabelige Fakultet, Københavns Universitet, København, Danmark, diskurs:7050              
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 Abstract: Introduction:
Neuropathic pain is a condition consisting of complex pathogenesis and the treatment today is primarily targeted on relief of symptoms. The glutamate transporter system is a relatively uninvestigated drug target for pain, but in neuropathic pain a reduction in the expression of glutamate transporters is observed, making this an interesting target. The glutamate transporters 1 (EAAT2/GLT-1) are responsible for >90 % of the glutamate clearance from the synaptic cleft. β-lactams, including ceftriaxone, has shown to be stimulators for the up regulation of the glutamate transporter 1.
Objectives:
The primary objective was to learn the procedure of Western blotting in order to be able to evaluate the expression of the glutamate transporter 1 in the spinal cord of CCI injured rat treated with 200 mg/kg ceftriaxone. A secondary objective was to investigate whether extracellular glutamate concentration in the cerebrospinal fluid of the same rat could be used to support the effect of the CCI surgery and the following ceftriaxone treatment.
Methods:
All rats underwent a CCI surgery to model neuropathy in man. The rats were afterwards treated with 200 mg/kg ceftriaxone for 10 days. The cerebrospinal fluid and spinal cord were isolated at different stages of the treatment. The expression of glutamate transporter 1 was investigated using Western blotting, and the glutamate concentration in the cerebrospinal fluid was determined by HPLC analysis.
Results:
The western blot analysis indicated the CCI surgery induced a down regulation of the glutamate transporter 1. The reduction was most outspoken for the ipsilateral part of the spinal cord. Ceftriaxone reversed the reduction of the GLT-1. A significant difference was seen comparing the ipsilateral part of the spinal cords of the non-treated CCI injured rats with the CCI inured rats which had been treated with ceftriaxone for ten days (P<0.01). Following re-down regulation was initiated two days after ended treatment. The HPLC analysis of the cerebrospinal fluid did not show any significant changes in the glutamate concentrations pre-treatment, during treatment or post-treatment.
Conclusion:
It was confirmed that the CCI surgery induced a knock down in the glutamate transporter 1. Ceftriaxone demonstrated a positive effect on the glutamate transporter 1 expression after the expression was reduced by the CCI surgery. The effect was not permanent and the expression decreased from day 2 post-treatment. The time of full decline of the effect of ceftriaxone was not elucidated. The concentration of glutamate concentration in the cerebrospinal fluid did not prove to function as an additive proof to Western blotting of the down and up regulation of the glutamate transporter 1.
 Abstract: Introduktion:
Neuropatisk smerte er en lidelse bestående af kompleks patogenese. Nuværende behandlingstilbud består primært af symptombehandling. Glutamattransportør-systemet er relativt uudforsket som mål for smertebehandling. En reduktion i ekspressionen i glutamattransportørene er observeret i neuropatisk smerte, hvilket gør dette system et interessant mål. Glutamattransportør 1 (EAAT2/GLT-1) er ansvarlig for >90 % af clearance af glutamate i den synaptiske kløft. β-laktamer, inklusiv ceftriaxon, stimulerer en opregulering af glutamattransportør 1.
Formål:
Hovedformålet var at lære proceduren for Western blotting, så det var muligt at evaluere ekspressionen af glutamattransportør 1 i rygmarven på CCI-opererede rotter, der var behandlet med 200 mg/kg ceftriaxon. Et sekundært formål var at undersøge om den ekstracellulære glutamatkoncentration i cerebrospinalvæsken på de samme rotter kunne anvendes til at supporte effekten af CCI-operationen og den følgende behandling med ceftriaxon.
Metode:
Alle rotterne blev CCI-opereret for at modellere neuropatisk smerte i mennesker. Rotterne blev efterfølgende behandlet med 200 mg/kg ceftriaxon i 10 dage. Cerebrospinalvæsken og rygmarvene blev udtaget på forskellige tidspunkter i forløbet. Ekspressionen af glutamattransportør 1 blev analyseret via Western blotting, og glutamatkoncentrationen blev analyseret vha. HPLC-analyse.
Resultater:
Analysen fra Western blotting viste, at CCI-operationen resulterede I en nedregulering i ekspressionen af glutamattransportør 1. Reduktionen var mest udtalt i den ipsilaterale side af rygmarven. Ceftriaxon vendte denne reduktion. Der blev observeret signifikant forskel mellem de ipsilaterale dele af rygmarvene hos ubehandlede, CCI-opererede rotters og CCI-opererede rotter behandlet med ceftriaxon i 10 dage (P<0,01). Effekten fortog fra anden dag efter endt behandling. HPLC-analysen af cerebrospinalvæsken viste ingen signifikante ændringer i glutamatkoncentrationerne før, under eller efter behandlingen.
Konklusion:
Det blev bekræftet, at CCI-operationen førte til en reduktion i ekspressionen af glutamattransportør 1. Ceftriaxon viste sig at have en positiv effekt på ekspressionen af glutamattransportør 1 efter ekspressionen først var reduceret af CCI-operationen. Effekten var ikke permanent, og var aftagende to dage efter endt behandling. Tiden for den totale remision af ceftriaxons effekt blev ikke undersøgt. Glutamatkoncentrationen i cerebrospinalvæsken viste sig ikke at være brugbar som forstærkende bevis til resultaterne fra Western blotting, der evaluerede ekspressionen af glutamattransportør 1.
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Copyright dato:
2013-11-11
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Bogmærk denne post: https://diskurs.kb.dk/item/diskurs:59127:7
 Type: Speciale
Alternativ titel: Effekten af ceftriaxon på ekpressionen af glutamattransportør GLT-1 i rygmarven samt på den ekstracellulære glutamatkoncentration i cerebrospinalvæsken i ”Chronic Constriction Injury” - rottemodellen for neuropatisk smerte.
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Detaljer

Sprog: English - eng
 Datoer: 2013-08-09
 Sider: 114 s.
 Publiceringsinfo: København : Københavns Universitet
 Indholdsfortegnelse: -
 Note: Farmaceutisk videnskab, cand. pharm. scient.
 Type: Speciale
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