前体脂肪细胞3T3-L1诱导分化条件的优化

利用L18(37)正交试验,考察了联合诱导时3-异丁基-1-甲基黄嘌呤(IBMX)、地塞米松和胰岛素用量以及联合诱导时间,胰岛素单独诱导时间以及胰岛素用量对3T3-L1前体脂肪细胞分化为成熟脂肪细胞的影响.极差分析的结果表明,各条件对3T3-L1前体脂肪细胞分化的影响程度由大到小排序为:IBMX、联合诱导时间、胰岛素单独作用量、联合诱导时胰岛素作用量、地塞米松、胰岛素单独作用时间;方差分析的结果表明,IBMX的诱导效应极显著,联合诱导时间和胰岛素单独作用量呈显著水平,而其他条件(联合诱导时胰岛素作用量、地塞米松和胰岛素以及单独作用时间)对结果的影响并不显著.3T3-L1前体脂肪细胞分化的最适诱导条件为:500~750μmol/L IBMX、0.25~1μmol/L地塞米松与1μg/mL胰岛素,在联合诱导72 h后,再用5~10μg/mL胰岛素诱导48 h.油红染色镜检结果显示,在该条件下前体脂肪细胞诱导分化率均高于联合诱导时胰岛素作用量、地塞米松和胰岛素以及单独作用时间.

Optimization of Induction Condition of 3T3-L1 Preadipocytes Differentiation

The present study investigated the influences of the concentration of 3-isobutyl-l-methylxanthine （IBMX）, dexamethasone and insulin during the combined induction stage （first stage） and the corresponding induction time, the induction time and concentration of insulin during the differentiation stage （second stage） on 3T3- L1 preadipocytes differentiation using Lls （37） orthogonal experiment. Range analyses showed that the impacts of those factors on results were （from high to low） ： IBMX, induction time of the first stage, insulin concentration during the second stage, insulin concentration dur ing the first stage, dexamethasone concentration and induction time of the second stage. Analysis of variance showed that IBMX played a highly significant influence on the process of 3T3-L1 preadipocytes differentiation, and the influence of induction time of the first stage and insulin concentration during the second stage were significant. In contrast, no significant difference was observed in the treatments of dexamethasone and insulin concentration. The optimal induction condition for differentiation of 3T3-Ll preadipocytes was presented as following： 500 - 750 μmol/L IBMX, 0. 25 - 1 μmol/L dexamethasone, and 1 μg/mL insulin for 72 h, and then 5 - 10 μg/mL insulin for 48 h. The result of red oil O staining showed that the differentiation rate of 3T3-L1 preadipocytes of the presented optimal condition was higher than that of the rest.