Effects Of Different Biotin’s Concentrations On Lipid Metabolism Of 3T3-L1 Adipocytes

This experiment was divided into two parts,cell induction and biotin intervention,with the object to study the effects of different biotin concentrations on the lipid metabolism of 3T3-L1 adipocytes.In part 1,3T3-L1 cells were induced differentiation into 3T3-L1 adipocytes.In part 2,different doses of biotin were added into cell culture medium as the trial group,in which the concentration of biotin in the culture medium were 0 μmol/L,0.2 μmol/L,0.5 μmol/L,and 1 μmol/L,respectively.Each group had 3 replicates,and each replicate had 2 parallel samples.Cell supernatants and adipocytes were harvested at 12 h,24 h,and 48 h and tested.The results of the experiment indicate that the survival rate of cultured 3T3-L1 cells tended to be stable after P6 generation;3T3-L1 adipocytes induced by the triple induction method had biological activity and can be used in subsequent experiments.At each time point,addition of biotin promoted triglyceride(TG)deposition and increased m RNA expression of Glucose Transporter-4 protein(GLUT-4),pyruvate kinase(PK),Acetyl-Co A Carboxylase 1(ACC1),fatty acids synthase(FAS),peroxisome proliferator-activated receptor γ(PPARγ),Perilipin A,adipose differentiation-related protein(ADRP),etc.,and decreased glycerol and leptin,and contents of tumor necrosis factor-α(TNF-α),interleukine-6(IL-6),and decreased the m RNA expression of hormone sensitive lipase(HSL)and adipose triglyceride lipase(ATGL).The concentration of biotin at 1 μmol/L significantly increased the m RNA expression of GLUT-4,PK,ACC1,PPARγ,Perilipin A and ADRP(P<0.01),and significantly decreased the release of glycerol and m RNA expression of ATGL(P<0.01).The concentration of biotin at 0.5 μmol/L biotin significantly increased the m RNA expression of HSL at 12 h(P<0.05)and 48 h(P<0.01).Conclusion: Addition of biotin increased the content of ADPN and the m RNA expression of FAS,ACC1,GLUT-4,PK,Perilipin A and ADRP in In vitro cultured 3T3-L1 adipocytes,and decreased the content of leptin,TNF-α,IL-6 and m RNA expression of HSL and ATGL.The concentration of 1 μmol/L was determined optimal concentration to promote the TG deposition and inhibit TG decomposition.