Immunohistochemical assessment of effectiveness of Carnitine Palmitoyl Transferase 1 (CPT1) blocking in rat model of experimental autoimmune encephalitis (EAE).
Author
Oklinski, Kirsten Egelund
Term
4. term
Publication year
2018
Submitted on
2018-12-21
Pages
33
Abstract
Baggrund: Multipel sklerose (MS) er en kronisk, betændelsespræget nervesygdom, men sygdommens præcise mekanismer er stadig uklare. En nyere hypotese peger på, at oxidativt stress i mitokondrier kan forstyrre cellers lipidmetabolisme. I dette arbejde undersøger vi etomoxir, der irreversibelt hæmmer enzymet carnitin palmitoyltransferase 1a (CPT1a), som indgår i cellens energiomsætning i mitokondrier. Hæmning af CPT1a forventes at tvinge cellen til at danne energi (ATP) fra andre kilder end normalt. Metode: Vi sammenlignede lillehjerne (cerebellum) og hjernestamme fra rotter behandlet med etomoxir, placebo eller ubehandlede raske kontroller. Ved hjælp af immunhistokemi med fluorescens- og peroxidbaserede farvninger målte vi myelin basic protein (MBP, en markør for myelin), CPT1a-ekspression samt tilstedeværelsen af antistofferne IgE og IgG. Resultaterne blev kvantitativt analyseret. Resultater: I lillehjernen var MBP-niveauet i etomoxir-behandlede rotter statistisk signifikant anderledes end i placebo- og raske kontrolgrupper, mens der ikke sås en signifikant forskel i hjernestammen. CPT1a-ekspressionen var signifikant lavere i etomoxir-gruppen end i placebo i både lillehjerne og hjernestamme, hvilket er foreneligt med en påvirkning af mitokondriernes energiomsætning. IgE-farvning sås generelt i både lillehjerne og hjernestamme, ofte lokaliseret i cellernes kerner uafhængigt af celletype. IgG-farvningen var stærkere i placebo-dyr og var tydeligst i strukturer, der typisk påvirkes ved neuronskade. Konklusion: Fundene tyder på, at etomoxir kan støtte myelinreparation i lillehjernen og samtidig ændre mitokondriernes energimetabolisme (indirekte vist ved nedreguleret CPT1a). De observerede IgE- og IgG-mønstre peger på en immunologisk komponent i de undersøgte hjerneområder.
Background: Multiple sclerosis (MS) is a chronic, inflammatory disease of the nervous system, but its exact mechanisms remain unclear. A recent hypothesis suggests that oxidative stress in mitochondria may disrupt cellular lipid metabolism. Here, we examine etomoxir, an irreversible inhibitor of carnitine palmitoyltransferase 1a (CPT1a), an enzyme involved in mitochondrial energy use. Blocking CPT1a is expected to force cells to generate energy (ATP) from alternative sources. Methods: We compared the cerebellum and brainstem of rats treated with etomoxir, placebo, or left as healthy controls. Using immunohistochemistry with fluorescent and peroxidase-based staining, we measured myelin basic protein (MBP, a marker of myelin), CPT1a expression, and the presence of the antibodies IgE and IgG. Results were quantitatively analyzed. Results: In the cerebellum, MBP levels in etomoxir-treated rats were statistically significantly different from those in placebo and healthy controls, whereas no significant difference was found in the brainstem. CPT1a expression was significantly lower in the etomoxir group than in placebo in both regions, consistent with an effect on mitochondrial energy metabolism. IgE staining was broadly observed in both regions, often localized to cell nuclei across cell types. IgG staining was stronger in placebo animals and most prominent in structures typically affected by neuronal injury. Conclusion: These findings suggest that etomoxir may support myelin repair in the cerebellum and shift mitochondrial energy metabolism (indirectly indicated by reduced CPT1a). The observed IgE and IgG patterns point to an immune component in the examined brain areas.
[This abstract was generated with the help of AI]
Keywords
multiple sclerose ; CPT1 ; EAE ; Hjernestamme ; dyremodel ; MBP
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