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目的:观察神经肽Y对3T3L1前脂肪细胞增殖和分化的影响,并对其机制进行初步探讨。方法:3T3-L1细胞由3-异丁基-1-甲基黄嘌呤(3-isobutyl-1-methylxanthine,IB-MX)、胰岛素和地塞米松联合诱导分化,2 d后将细胞分为空白对照组(未加任何诱导剂组)、经典诱导组(胰岛素诱导)和NPY干预组(10-8、10-9、10-10mol/L NPY)。培养的第7、12天用相差显微镜观察各组细胞形态的变化,培养第12天用油红O染色观察脂肪细胞分化程度。MTT检测细胞增殖。Western blot检测脂肪细胞分化相关基因过氧化物体增殖剂活化受体-γ(peroxisome proliferator-activated receptor gamma,PPAR-γ)、CAAT/增强子结合蛋白-α(CCAAT/enhancer-bind-ing protein-α,C/EBP-α)蛋白的表达。结果:10-8mol/L NPY能促进3T3-L1细胞的分化。10-9mol/L NPY,10-8mol/L NPY均能促进3T3-L1细胞增殖。10-8mol/L NPY增加C/EBPα、PPARγ的表达。结论:NPY促进3T3L1前脂肪细胞的增殖和分化,其机制可能与上调PPARγ、C/EBPα的表达有关。
Objective: To observe the effect of neuropeptide Y on the proliferation and differentiation of 3T3L1 preadipocytes, and to explore its mechanism. Methods: The 3T3-L1 cells were induced to differentiate by 3-isobutyl-1-methylxanthine (IB-MX), insulin and dexamethasone. After 2 days, the cells were divided into blank Control group (without any inducer), classic induction group (insulin induced) and NPY intervention group (10-8,10-9,10-10 mol / L NPY). On the 7th and 12th days of culture, the morphological changes of the cells in each group were observed by phase contrast microscopy. On the 12th day of culture, the adipocyte differentiation degree was observed by oil red O staining. MTT assay of cell proliferation. Western blot was used to detect the expression of adipocyte differentiation-related genes peroxisome proliferator-activated receptor gamma (PPAR-γ), CAAT / enhancer-bind-ing protein-α , C / EBP-α) protein expression. Results: 10-8mol / L NPY promoted the differentiation of 3T3-L1 cells. 10-9mol / L NPY, 10-8mol / L NPY can promote 3T3-L1 cell proliferation. 10-8 mol / L NPY increased the expression of C / EBPα and PPARγ. Conclusion: NPY can promote the proliferation and differentiation of 3T3L1 preadipocytes. The mechanism may be related to up-regulating the expression of PPARγ and C / EBPα.