Effects Of Metformin On MRNA Expression Of Vesicle-associated Membrane Protein In Differentiation Process Of 3T3-L1 Preadipocytes

Objective:To observe mRNA expression of v-SNAREs in differentiation process of 3T3-L1 preadipocytes,and to explore mechanism of metformin in GLUT4 transport.Method:3T3-L1 preadipocytes were cultured.The cells were induced when they were grown to confluence reaches 100% or over-fusion at.The cells were induced by fluid induced to differentiate, induction medium A was 3 days, B solution one day, alternately 3 times, a total of 12 days.In the induction process, were given different concentrations of metformin(0、0.5、1、2mmol/L) continued to intervene, the first 12 days to collect specimens. Induce fat by oil red O staining. Using RT-PCR technology to detect mRNA of AMPK、GLUT4、VAMP2、3、4、5、7、8 in adipocytes. Experimental data applications SPSS13.0 software for statistical analysis, differences between groups using single factor analysis of variance,multiple comparisons between groups using LSD test, test level taking α=0.05.Results:Using ANOVA,the results of AMPK、VAMP4、VAMP7 mRNA expression were not statistically significant.The results of GLUT4、VAMP2、3、5、8 mRNA expression were statistical significance, LSD test results: 1) Compared with group of 0mmol/L, GLUT4、VAMP2、3、8 mRNA levels in group of 0.5mmol/L were no significant difference(P>0.05);2) compared with group of 0mmol/L, GLUT4、VAMP2、3、8 mRNA levels increase in group of 1mmol/L,the difference reached statistical significance(P<0.05); 3) compared with group of 1mmol/L,GLUT4 、 VAMP2 、 3 、 8 mRNA levels increase in group of2mmol/L,the difference reached statistical significance(P<0.05).Conclusion:Metformin may act on pre-adipocyte differentiation process.In 1mmol/L concentration range, with increasing concentrations of metformin, may increase mRNA expression of GLUT4 、 VAMP2 、 3 、 8 in adipocyte,with a certain degree of dose-dependently.1mmol/L is probably the largest concentration in 3T3-L1 preadipocytes for metformin. Metformin may exert on the GLUT4 transport in adipocyte regulated by adjusting the v-SNAREs protein family.When metformin concentration is 2mmol/L,metformin will decrease mRNA expression of GLUT4、VAMP2、3、8. In this study provide a theoretical basis for further exploration of metformin in GLUT4 translocation to the specific mechanism of action.