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Objective: Aquaporin adipose (AQPap) glycerol operation located on the fat cell membrane protein, which is responsible for the glycerol transporter to participate in the maintenance of blood glucose in the extracellular lipolysis. Appropriate conditions, cell differentiation of 3T3-L1 preadipocytes into mature fat cells to observe the regulatory role of hormones, nutrients, cholesterol lowering agents the stimulation of AQPap expression, and to study of human adipose the organization AQPap expression changes in the amount and weight relationship , to explore AQPap expression regulating mechanism and its role in obesity, diabetes pathogenesis. Methods: 1, 0.5mMIMX 1μM dexamethasone 5μg/ml insulin-induced induction of differentiated cells and oil red O staining. 2, the use of semi-quantitative RT-PCR and Western blotting, from gene and protein expression levels of hormones (insulin, epinephrine, dexamethasone, and IGF-1), nutrients (glucose, glycerol), lipid-lowering drugs (non-promise Bate, lovastatin) expression of 3T3-L1 adipocytes cells AQPap observed the different weight subjects adipose tissue AQPap expression changes. Results: 1, 3T3-L1 cells differentiated from spindle gradually becomes circular, and increased cell volume, cytoplasmic lipid droplets within nine days, more than 95% of the field of view in the cells to differentiate into typical mature fat cells. Oil Red O staining the nuclei of mature fat cells as blue intracytoplasmic fat droplets were stained orange-red. 2, as compared with the control group, to give the adrenergic stimulation of the different concentrations of the differentiated and mature 3T3-L1 cells, the the AQPap mRNA expression level of no significant changes (p> 0.05). -9 M insulin-stimulated cells 6 hours can AQPap gene expression was decreased 16%, 10 -6 M islet eleven to one prime declined by more than 50% 48 hours after the AQPap protein levels compared with the control group declined by 64o. IGF Mateo also reduce of AQPap the gene expression, but the the same concentrations door - 'M) stimulus, the role of the IGFJ weaker than that of insulin (P, 0 knife 5). Dexamethasone of AQPcymRNA expression had no significant effect, however, while giving 10'M of dexamethasone and insulin alone 10 - 'M insulin phase t Bu AQPap mRNA expression levels increased (p <0.05 ). Glucose concentration increased expression AQPap m Chau A significant increase (p, 0 * 5), 16. the smM glucose-stimulated 48 hours, AQPap mRNA expression level was about three times the control group (glucose concentration of 5,6 mM); increased protein levels nearly doubled. Glycerol stimulate AQPap mRNA expression was not significantly changed. Lipid-lowering drug fenofibrate, lovastatin 3T3 workers l the fat cells AQPap gene expression (P> O no direct impact on the knives 5). , Obesity and overweight of adipose tissue AQPap gene expression were significantly higher than the normal weight group Zhongzhou difference between 05X overweight and 8 bar fat group. Western blotting results show that, compared with the normal weight group, obese group only of adipose tissue AQPap protein expression levels increased significantly (p Sichuan knife 5), no significant differences in other groups. Conclusion: 1, oil red O staining, the induced differentiation program enables uniform cell differentiation, cell differentiation, high success rate,%% fusiform precursor cells into large round white white mature fat containing lipid droplets fine month. 2, insulin from 3T3 soil of fat cells AQPaP the expression inhibited dose-and time-dependent. IG = 1 also suppressed I effect, but more weakly than insulin. Adrenaline. Species lipolytic hormones, no effect it AQP rapping mRNA expression. The dexamethasone able antagonist downward effect of insulin on the the 3T3 soil the fat cells AQpap gene expression. The high glucose AQPap mRNA and protein expression is significantly enhanced. Glycerol and lipid-lowering drug fenofibrate, lovastatin no direct impact on the the 3T3 small l the fat cells AQPaP gene expression. 3, the adipose tissue of human obese patients AQPaP expression is enhanced; weight change and AQPap expression is abnormal to see a close relationship. } Yishan, Beijing K village: M Bu Dissertation
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