| Summary: | 碩士 === 國立臺灣大學 === 食品科技研究所 === 105 === Diabetes becomes a global health issue as a result of the shifting of dietary habits. Previously, the population of patients was seen mainly in the middle-aged and elderly, but it is getting more frequently among young people. Glucose serves as energy source for most cells. Blood sugar rises after a meal and is mainly absorbed by skeletal muscles and adipose tissues. Glucose transporter 4 (GLUT4), the downstream effector of insulin, takes an important role in glucose uptake. Insulin resistance would cause defective insulin response in target cells, leading to hyperglycemia. However, approaches in measuring glucose uptake are considered to be time consuming, expensive and not easy to operate. Also, the results may be affected by other metabolic pathways in cells, and is incompatible with the actual physiological environments. According to these disadvantages, we attempted to establish a more rapid and intuitive method to measure glucose uptake through the construction of a cell platform. Based on this reason, we transfected a recombinant plasmid into CHO-K1 to produce GLUT4-eGFP recombinant protein. Antibiotics selection was carried out to exclude the non-transfected cells. The fluorescence intensity was analyzed to estimate the stability and expression efficiency of this fusion protein. In addition, the GLUT4-eGFP recombinant protein is responsive upon insulin stimulation, and the fluorescence was detected by Total internal reflection fluorescent microscope (TIRFM) afterwards. The result shows that our detection method is correlated to glucose uptake and is more efficient than the traditional methods, indicating that this cell platform can be applied to screen potential compounds with the ability to increase cellular glucose uptake.
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