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The Effect of Insulin on Glucose Metabolism

Stoffers, A.E. (2015) The Effect of Insulin on Glucose Metabolism. Master's Thesis / Essay, Biology.

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Development of T2DM is usually preceded by an increased metabolic demand for insulin due to insulin resistance in several tissues. We would like to elucidate the effect of insulin on glucose metabolism under insulin sensitive and resistant conditions, using TNF-ɑ to induce insulin resistance in differentiated 3T3L1 cells. As a pilot study first in vitro and in vivo NAD(P)H-linked enzyme assays were set up for the glycolytic enzymes. Overall the activity of the enzymes measured under in vitro conditions was slightly higher. Using 2.5 nM TNF-α and 100 nM insulin, four cell conditions were made: without TNF-α without insulin, without TNF-α with insulin, with TNF-α without insulin and with TNF-α with insulin. In the four conditions differences in glucose-consumption, lactate-production, pyruvate concentration and glycolytic enzymes activities were determined. Glucose consumption was measured over 24 hours. TNF-α did not induce insulin resistance. However, it did decrease glucose consumption of cells. Insulin did increase glucose consumption. Lactate and pyruvate were measured in the medium at the 24 hour time point. Lactate production was increased by insulin and TNF-α, but no effects were seen on pyruvate concentration. The in vitro assays were used to measure enzyme activities. Insulin increased the activity of the enzymes, TNF-α had no effect on activity. To conclude, no insulin resistance was induced. However, TNF-α still had an effect on the glucose metabolism. It caused decreased glucose consumption and more glucose to be converted to lactate. The mechanism behind this is unknown. A remarkable effect of insulin was the increased enzyme activities. A lot of research still has to be done to optimize the conditions of the experiment. In the future, in vivo conditions should be used and with these results a quantitative computer model of the glucose metabolism can be developed.

Item Type: Thesis (Master's Thesis / Essay)
Degree programme: Biology
Thesis type: Master's Thesis / Essay
Language: English
Date Deposited: 15 Feb 2018 08:04
Last Modified: 15 Feb 2018 08:04

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