faculteit: "FNWI" en publicatiejaar: "2011"
| Auteur||DEWI VAN HARSKAMP|
|Titel||DEVELOPMENT OF A METHOD TO MEASURE VLDL SYNTHESIS RATES|
|Begeleiders||M.T. ACKERMANS, W.TH. KOK|
|Faculteit||Faculteit der Natuurwetenschappen, Wiskunde en Informatica|
|Opleiding||FNWI MSc Chemistry|
|Samenvatting||Obesity is becoming a major health problem. It is associated with many diseases, amongst others the metabolic syndrome. A current research project at the AMC aims on elucidating the significance of the brain with respect to lipid metabolism in the liver, as an increased production of lipids is associated with the metabolic syndrome.
Lipids synthesized in the liver are secreted within Very Low Density Lipoprotein (VLDL). This protein belongs to a class of proteins, the lipoproteins, that is responsible for the transport of lipids through the blood. For research on the influence from the brain on the liver, a method has to be designed that can measure VLDL synthesis rates. Until now, this rate is measured by administering tyloxapol (triton WR1339), which inhibits LPL. However, this method presents some drawbacks.
The method to be developed during this research project will involve the infusion of stable isotope labelled d5-glycerol into subjects (rats). This labelled glycerol will be incorporated in triglycerides in the liver and secreted within VLDL. For the isolation of VLDL from blood plasma, AF4, FPLC, and ultracentrifugation are looked into. For the subsequent hydrolysis of triglycerides in the obtained VLDL fractions chemical and enzymatic hydrolysis are considered.
The resulting glycerol will be analysed by GC-MS to obtain Tracer-to-Tracee Ratios (TTR). These will be plotted against time, and from this graphs, the VLDL synthesis rate can be determined.
Unfortunately, the aim for this research project is not achieved. The bottleneck appeared to be the isolation of VLDL in such a way that free glycerol from the rat plasma is not present in the VLDL fraction. This undesired free glycerol will be measured during GC-MS analysis and affects the resulting TTR. However, for human subjects, there is a procedure that isolates VLDL without free glycerol from the plasma present in the fractions. Unfortunately, this procedure needs more
plasma than can be obtained from rats at subsequent time points.
Human VLDL fractions obtained by this ultracentrifugation procedure were used to prove it is possible to obtain a TTR curve that is similar to those in literature, as to prove the method designed during this project gives the desired results.|
|Soort document|| scriptie master|
Gebruik dit adres om naar deze pagina te linken: http://dare.uva.nl/scriptie/384472
Vraag/opmerking over dit recordMail aan een collega
Toevoegen aan bewaarset