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目的观察游离脂肪酸对3T3-L1细胞分化及insig2 mRNA基因表达的影响,进一步了解影响3T3-L1细胞分化成熟及insig2表达的因素,探讨insig2基因在脂肪细胞分化与代谢中的作用。方法将3T3-L1细胞分为高脂组与对照组,分别置于低糖高脂与低糖环境中分化培养12 d,油红O染色计算染色阳性细胞面积,RT-PCR法测定in-sig2、ap2 mRNA的表达。结果分化培养12 d后,油红O染色显示两组细胞均得到明显分化,但高脂组油红O染色阳性细胞面积较对照组明显升高(P<0.05)。分化后,两组细胞ap2、insig2表达水平均较分化前明显升高(P>0.05),且高脂组较对照组上调明显(P<0.05)。结论游离脂肪酸可促进3T3-L1细胞分化成熟与insig2表达;insig2在细胞内的表达水平可能与脂肪细胞成熟及分化程度相关。
Objective To investigate the effects of free fatty acids on the differentiation of 3T3-L1 cells and the expression of insig2 mRNA, to further understand the factors that affect the differentiation and insig2 expression of 3T3-L1 cells, and to explore the role of insig2 in adipocyte differentiation and metabolism. Methods 3T3-L1 cells were divided into hyperlipidemia group and control group. The cells were cultured in low glucose, high fat and low glucose for 12 days. The area of positive cells was calculated by oil red O staining. The expression of in-sig2, ap2 mRNA expression. Results After differentiation for 12 days, oil red O staining showed that both groups of cells differentiated significantly, but the area of oil red O staining positive cells in hyperlipidemia group was significantly higher than that in control group (P <0.05). After differentiation, the expression levels of ap2 and insig2 in both groups were significantly higher than those before differentiation (P> 0.05), and were significantly up-regulated in hyperlipidemia group compared with the control group (P <0.05). Conclusion Free fatty acids can promote the differentiation and maturation of 3T3-L1 cells and insig2 expression. The expression of insig2 in cells may be related to the maturation and differentiation of adipocytes.