Abstract: 【Objective】 In this study, the purpose is to induce 3T3-L1 cells to differentiate into 3T3-L1 adipocytes and to study the effects of various biotin levels on the transcription and expression of adipose synthase related genes in 3T3-L1 adipocytes.【Method】3T3-L1 cells were induced to differentiate into 3T3-L1 adipocytes by triplex induction. When 3T3-L1 adipocytes were dense, they were treated by 0 μmol/L biotin (control group), 0.2, 0.5 and 1 μmol/L The relative expression of competitive PKmRNA, GLUT-4 mRNA, FAS mRNA and ACC1 mRNA in cell supernatant was detected at 12h, 24h and 48h, respectively.【Result】Results showed that when the test time was 12 h: GLUT-4 mRNA, PK mRNA and ACC1 mRNA relative expression quantity of experimental groups was significantly (P＜0.01) higher than that of control group, FAS mRNA relative expression quantities of 0.5 μmol/L group and 1 μmol/L group were significantly (P＜0.01) higher than that of control group. When the test time was 24 h: GLUT-4 mRNA and PK mRNA relative expression quantity of 1 μmol/L group was significantly (P＜0.05) higher than that of control group, ACC1 mRNA relative expression quantity of 1 μmol/L group was extremely significantly higher (P＜0.01) than that of control group,ACC1 mRNA relative expression quantity of 0.2 μmol/L group and 0.5 μmol/L group was significantly higher (P＜0.05) than that of control group. When the test time was 48 h, GLUT-4 mRNA relative expression quantity of 1 μmol/L group was significantly (P＜0.05) lower than that of control group, PK mRNA relative expression quantity of 1 μmol/L group was extremely significantly (P＜0.01) higher than that of control group, FAS mRNA relative expression quantity of 0.5 μmol/L group was significantly (P＜0.05) higher than that of 1 μmol/L group, FAS mRNA relative expression quantity of 1 μmol/L group was significantly (P＜0.05) higher than that of control group, ACC1 mRNA relative expression quantity of 1 μmol/L group was extremely significantly (P＜0.01) higher than that of other groups.【Conclusion】The addition of biotin can inhibit the adipogenesis through promoting the relative expression of GLUT-4, PK, FAS and ACC1 mRNA in adipocytes. Moreover, when the concentration is equal to 1 μmol/L, the effect is optimal to GLUT-4, PK and ACC1 mRNA, and when the concentration is equal to 0.5 μmol/L, the effect is optimal to FAS mRNA.