Effect Of A-FABP Silencing On Triglycerides Production And Adiponectin Secretion In 3T3-l1 Adipocyte

Objective:To construct the MicroRNA expression vector of A-FABP gene, observing the effect of RNA interference on A-FABP gene expression and Synthesis of triglycerides and adiponectin secretion in 3T3-L1 Adipocyte.Methods: Constructed the MicroRNA expression vector of A-FABP gene, LipofectamineTM2000 transfected the constructed recombinant plasmid into 3T3-L1 adipocyte.Expression of A-FABP mRNA and protein were detected by reverse transcription polymerase chain reaction(RT-PCR)and Western blotting respectively. The induction of matured 3T3-L1 adipocytes were incubated for 24h with 0 1mmol/ L free fatty acids (FFA) Measured their concentration of triglycerides and adiponectin by ELISA Kit Established cell model of A-FABP gene which be silenced,and incubate with 0.5mmol / L FFA ,divided into 4 groupcontrol 0.5mmol/LFFA RNAi RNAi +0.5mmol/LFFA the concentration of triglycerides measured by enzyme assay and adiponectin determined by ELISA Kit. A-FABP and adiponectin mRNA Expression levels were avalued by RT-PCR.Results:1.Designed and constructed the targeting microRNA A-FABP gene expression vector, transfected it into 3T3-L1 adipocyte, inhibited the A-FABP mRNA and protein expression significantly. 2. With the fatty acid concentration increased, triglyceride concentration increased (P <0.05), while adiponectin secretion decreased in matured 3T3-L1 adipocytes (P <0.05). 3.The triglyceride concentration significantly decreased and the adiponectin secretion and mRNA expression significantly increased in RNA interference group compared with control group P < 0.05 .Conclusions: A-FABP gene Silenced can decreased triglyceride synthesis,but increased adiponectin secretion in 3T3-L1 adipocyte,the effects might be a new target for preventing and treating atherosclerosis and diabetes.