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Effekten av 1- metylnikotinamid på glukoneogenes hos humana hepatocyter. En metodoptimeringsstudie
Kristianstad University, School of Education and Environment.
2015 (Swedish)Independent thesis Basic level (university diploma), 10 credits / 15 HE creditsStudent thesis
Abstract [sv]

Nikotinamid-N-metyltransferase (NNMT) är ett enzym som katalyserar reaktionen där nikotinamid (amidformen av vitamin B3) metyleras till 1-metylnikotinamid (MNA). Enzymet NNMT och dess produkt MNA har bevisats ha en koppling till insulinresistens, fetma och typ 2 diabetes (T2D). Frågeställningen i den här pilotstudien inriktar sig på 1-metylnikotinamid (MNA) och dess betydelse för glukosmetabolismen, med hypotesen att den ökar glukoneogenesen hos hepatocyter. Syftet med studien var att optimera en metod för att studera hepatocyternas glukosproduktion in vitro samt undersöka MNAs effekt på produktionen. I arbetet undersöktes två olika metoder utformade för att stimulera igång glukoneogenesen hos den hepatocarcinoma cellinjen HUH7. Effekten av tre olika MNA koncentrationen (0,5, 1 och 10mM) på glukoneogensen har undersökt och glukosmätning i cellmedier utfördes med en hexokinas assay. Hypotesen om MNA har inte kunnat bekräftas eller uteslutas utifrån de erhållna resultaten. Ingen av metoderna var optimala för att undersöka glukoneogenesen hos hepatocyter in vitro utan eller med stimulering av 1-metylnikotinamid. Metoderna behöver optimeras vidare vad gäller cellernas odlingsförhållanden och även val av assay för att mäta glukos i cellmedier.

Abstract [en]

The enzyme Nicotinamide-N-methyltransferase (NNMT) catalyzes the reaction in which nicotinamide (the amide form of vitamin B3) is methylated to 1-methylnicotinamide (MNA).  NNMT and its product MNA has previously been proven to be associated with insulin resistance, obesity and type 2 diabetes (T2D). This pilot study focuses on MNA and its importance for glucose metabolism, with the hypothesis that it increases gluconeogenesis in hepatocytes. The purpose of the study was to optimize a method to investigate the effect of MNA on gluconeogenesis. Two different methods were designed to stimulate gluconeogenesis in a HUH7 hepatocarcinoma cell line. The effect of three different MNA concentration (0.5, 1 and 10mM) were examined and glucose measurement in cell media was performed using a hexokinase assay. The results doesn’t confirm nor reject the MNA hypothesis, for the reason that none of the used methods were competent for this type of analyses. The methods needs to be further optimized, in which there shall be better cell culture conditions and also a better choice of a glucose assay.  

Place, publisher, year, edition, pages
2015. , 17 p.
Keyword [en]
HUH7, hepatocytes, NNMT, MNA, gluconeogenesis, glucose output, type 2 diabetes
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:hkr:diva-14212OAI: oai:DiVA.org:hkr-14212DiVA: diva2:828064
Subject / course
Biomedical Laboratory Science
Educational program
Bachelor of Science in Biomedical Laboratory Science
Supervisors
Examiners
Available from: 2015-08-07 Created: 2015-06-29 Last updated: 2015-08-07Bibliographically approved

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CiteExportLink to record
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Citation style
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