3T3-L1前体脂肪细胞诱导分化方法优化的初步探讨

目的 观察不同诱导剂对3T3-L1前体脂肪细胞诱导分化的作用。方法 在经典鸡尾酒诱导剂成分基础上，根据不同诱导剂成分，将3T3-L1前体脂肪细胞分为4组：（1）对照组；（2）罗格列酮干预组；（3）吲哚美辛干预组；（4）罗格列酮和吲哚美辛联合干预组。在诱导前体脂肪细胞分化过程中，通过显微镜观察细胞形态学变化、油红O染色以及检测胞内和胞外三酰甘油含量，评价各组成脂效率。结果 罗格列酮和吲哚美辛单独干预组及联合干预组前体脂肪细胞诱导过程中更早出现脂滴，且形成的脂滴也较对照组多，而油红O染色进一步证实以上结果。采用Image Pro Plus 6.0软件统计经油红O染色后的脂滴，单个脂滴的平均直径、平均面积以及最大直径，罗格列酮干预组和联合干预组均大于对照组，而吲哚美辛干预组与对照组比较，差异无统计学意义（P>0.05）。三酰甘油含量检测，胞外三酰甘油含量4组比较，差异无统计学意义，但胞内三酰甘油含量联合干预组高于对照组[（0.77±0.07）mmol/g vs（0.19±0.02）mmol/g，P<0.05]。结论 优化经典鸡尾酒诱导方法，在原成分基础上加用过氧化物酶体增生因子活化γ型受体（peroxisome proliferator-activated receptor，PPAR-γ）激动剂罗格列酮，可显著提高3T3-L1前体脂肪细胞的诱导效率。

Optimization of induction and differentiation of 3T3-L1 preadipocyte

Objective To optimize the methods of the induction and differentiation of 3T3-L1 preadipocytes. Methods Based on the "cocktail" method, 3T3-L1 preadipocytes were divided into four groups according to the differentiation medium A:(1) Control group; (2) Ros:rosiglitazone group; (3) Indo:rosiglitazone group; (4) Ros+Indo:rosiglitazone+indometacin combination group. During the differentiation, cell morphology was observed through microscope. Oil red O was stained and quantified. The intracellular and extracellular triglyceride (TG) content was measured via TG GPO-PAP enzymatic kit. Results The differentiation rates were higher in group "Ros", "Indo" and "Ros+Indo", with earlier adipogenesis and more lipid droplets, which was further demonstrated in oil red O staining. In the evaluation of one simple lipid droplet with three parameters:mean diameter, mean area and max diameter, the droplet in group "Ros", "Indo" and "Ros+Indo" were much bigger than that in control group, while there was no statistically significance between control and "Indo" group. Regarding TG content, there was no significant difference among four groups in extracellular TG, but intracellular TG was higher in "Ros+Indo" group than that in control group (0.77±0.07 mmol/g vs 0.19±0.02 mmol/g, P<0.05). Conclusion Based on the classical "cocktail" method, PPAR-γ agonist rosiglitazone significantly improves the differentiation rate of 3T3-L1 preadipocytes.