Title page
Acknowledgement
Contents
Abstract 13
I. Introduction 15
II. Materials and Methods 22
II. 1. Materials 22
II. 2. Cell culture and differentiation 23
II. 3. Assay of Cell proliferation 24
II. 4. Flow Cytometry 24
II. 5. Oil red O stain 25
II. 6. Total RNA isolation and Semi-quantitative reverse transcriptase polymerase reaction (Semi RT-PCR) 26
II. 7. Statistical analysis 27
III. Results and Discussion 28
III. 1. Effect of fucoidan on XTT cell proliferation 28
III. 2. Cell-cycle progression with flow cytometric analysis 28
III. 3. Effects of fucoidan on intracellular lipid accumulation in 3T3-L1 29
III. 4. Effect of fucoidan on adipocyte differentiation 30
III. 5. Time course of C/EBPα expression in differentiating 3T3-L1 cells and effect of fucoidan 32
III. 6. Effect of fucoidan on PPARγ gene expression 33
III. 7. Effect of fucoidan on GLUT4 gene expression 34
References 51
Abstract in Korean 58
Table 1. In the last few decades the terminology for classifying the subtypes of diabetes has been decided as following Type 1 diabetes mellitus versus Type 2 diabetes mellitus. 37
Table 2. Currently available pharmacological agents for the treatment of Type 2 Diabetes Mellitus. 38
Table 3. Mammalian passive glucose transporters 39
Table 4. Primer sequences for RT-PCR analysis. 40
Figure 1. Through pathogenesis of the complication of insulin dependent diabetes mellitus (IDDM, T1DM) and non insulin dependent diabetes mellitus (NIDDM, T2DM). (John, 1992) 41
Figure 2. Morphology of 3T3-L1. (A) Preadipocytes fibroblast shape (B) Post-confluent 42
Figure 3. Stage in the adipocyte development program illustration the events and temporal expression of various preadipocyte and adipocyte markers. (Lane MD, 1995) 43
Figure 4. The structure of fucoidans (Top) F-fucoidan, (Middle) U-fucoidan, (Bottom) G-fucoidan. 44
Figure 5. Cell proliferation (3T3-L1 mouse adipocyte cell line) was determined by XTT assay to evaluate the cytotoxic effect of different fucoidan concentration (0, 10, 50, 100, and 45
Figure 6. The Cell-cycle progression was assessed with flow cytometric analysis of DNA staining with propidium iodide. (A) The result of the Preadipocyte gating as histogram and 46
Figure 7. Fucoidan inhibit adipogenic differentiation of 3T3-L1 preadipocytes. (Left) 3T3-L1 cells were induced to differentiate in 60 mm plates for 8 day as described in Materials 47
Figure 8. Effect of fucoidan on C/EBPα gene expression. (Top) Gel electrophoresis normalsation data of the expression of C/EBPα during 3T3-L1 adipocyte differentiation. (Bottom) 48
Figure 9. Effect of fucoidan on PPARγ gene expression. Legends are the same as Figure 8. 49
Figure 10. Effect of fucoidan on GLUT4 gene expression. Legends are the same as Figure 8. 50