| Mouse3T3-L1Preadipocytes were induced to differentiate into adipocytes.Then, adipocytes Insulin resistance(IR) model were created by high glucose and high High method.Our objective was to investigate the the effect of t-AUCB(trans--[4-(3-adamantan-1-y1-ureido)-cyclohexyl]-benzoic acid), a selective soluble epoxide hydrolase inhibitors, on glucose uptake and tumor necrosis factor-a (TNF-α) and adiponectin secretion and glucose transporter-4(GLUT-4) mRNA, protein expression in Insulin resistance adipocytes.Methods1.Mouse3T3-L1Preadipocytes were induced to differentiate into adipocytes;2. adipocyte insulin resistance model building:adipocytes were cultured with high glucose and high insulin method, and the model was identified through detection glucose uptake of adipocytes cultured;3. The adipocytes modeled after divided into5groups (including adipocytes for about2×104Cells), of which one was the control group, and the remaining four groups were the intervention groups, respectively, using different concentrations (0,1,10,50,100μmol/L)) t-AUCB intervention cultured24h, at the same time, take the common culture of fat cells as the control group. Supernatants were extracted, collected adipocytes;4. The concentration of glucose in the supernatant of each group was determined by glucose oxidase-peroxidase method;5.The levels of adiponectin and TNF-α in the supernatant were Measured by ELISA.6.GLUT4protein were Measured by Western Blot and GLUT4mRNA by Real time-PCR. Results1.3T3-L1preadipocytes were induced to differentiate into mature adipocytes. After oil red O staining, we can found that the fat droplets in mature adipocytes weres dyed orange Under the light microscope.2. There are significant differences between the glucose uptake rate of adipocytes cultured by high glucose and high insulin and cultured by general culture medium.(3±1.9%VS40±1.2%, P<0.05).3. After using1,10,50,100μmol/L of t-AUCB intervention, the glucose uptake rate of adipocyte respectively was7±1.5%,17±1.2%,32±1.6%,36±0.9%, significantly higher than the control group (3±1.4%), the difference was significant (all P<0.05), and showed a dose-dependent manner (all P<0.05). Blank control group compared with control group also significantly different (42±1.7%VS3±1.4%, P<0.05).4. After using1,10,50,100μmol/L of t-AUCB intervention,the levels of the supernatants adiponectin Concentration of adipocyte respectively was,0.64±0.07,1.30±0.11,1.66±0.05,2.33±0.04ng/mL, were significantly higher than control group (0.33±0.02), the difference was significant (all P<0.05), and showed a dose-dependent manner (all P<0.05). Blank control group compared with control group also significantly different (3.04±0.07VS0.33±0.02, P <0.05).5. After using1,10,50,100μmol/L of t-AUCB intervention, the levels of the supernatants TNF-a Concentration of adipocyte respectively were163.30±3.24,125.52±2.08,93.93±2.62,49.65±1.20pg/mL, were significantly lower than the control group (182.34±4.89pg/mL), the difference was significant (all P<0.05), and showed a dose-dependent manner (all P<0.05). Blank control group compared with control group also significantly different (8.06±1.80VS182.34±4.89pg/mL, P<0.05)6. After using1,10,50,100μmol/t-AUCB intervention, the RQ values of GLUT-4mRNA expression of adipocyte respectively was1.28±0.41,2.00±0.08,2.42±0.12,3.41±0.21, significantly higher than the control group (1.12± 0.18), the difference was significant (all P<0.05), and showed a dose-dependent manner (all P<0.05). Blank control group compared with control group also significantly different (3.67±0.26VS1.12±0.18, P<0.05).7. After using1,10,50,100μmol/L of t-AUCB intervention, the GLUT-4protein optical density (GLUT-4/GAPDH) of adipocyte respectively were0.329±0.118,0.472±0.123,0.623±0.172,0.756±0.112, significantly higher than the control group (0.312±0.115), the difference was significant (all P<0.05), and dose-dependent manner (all P<0.05). Blank control group compared with control group also significantly different (0.810±0.121VS0.312±0.115, P <0.05).Conclusions1. Epoxide hydrolase inhibitors t-AUCB may promote the GLUT-4protein expression, and glucose uptake in Insulin resistance Adipocytes By a way of dose-dependent manner, which improved the insulin resistance in Insulin resistance adipocytes.2. Epoxide hydrolase inhibitors t-AUCB may increase the secretion of adiponectin while reducing the secretion of TNF-a levels, By a way of dose-dependent manner in Insulin resistance Adipocytes,which improved Insulin resistance adipocytes secretory function. |
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