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目的:观察齐墩果酸(oleanolic acid,OA)对脂肪细胞分化和糖脂代谢的影响并探讨其机制。方法:在诱导脂肪分化时用OA干预,油红O染色后,通过比色法分析脂肪细胞的分化程度。采用葡萄糖氧化酶法检测OA对脂肪细胞葡萄糖消耗的影响;采用酶联免疫吸附法检测上清液中的游离脂肪酸浓度;采用RT-PCR法检测脂肪细胞PPAR-γ和GLUT-4 mRNA的表达。结果:与溶媒对照组比较,OA能显著增加脂肪细胞葡萄糖消耗量(P<0.05或P<0.01),在1~30μmol/L浓度范围时,其作用随着剂量增加显著增强;OA能促进前脂肪细胞的分化,在3~30μmol/L浓度范围内,OA组油红O染液光密度值均显著高于溶媒对照组(P<0.05或P<0.01)。同时,OA还能减少游离脂肪酸的产生,其30μmol/L剂量时游离脂肪酸的含量只有溶媒对照组的47.2%,显著低于溶媒对照组(P<0.01)。与溶媒对照组比较,30μmol/L OA能显著增加脂肪细胞PPAR-γ及GLUT-4 mRNA的表达(P<0.05)。结论:OA能促进脂肪细胞分化和葡萄糖的消耗,减少游离脂肪酸的产生,其机制可能与上调脂肪细胞PPAR-γ和GLUT-4 mRNA的表达、增加脂肪细胞对胰岛素的敏感性有关。
Objective: To observe the effect of oleanolic acid (OA) on adipocyte differentiation and glycolipid metabolism and to explore its mechanism. Methods: In the induction of fat differentiation with OA intervention, oil red O staining, by colorimetric analysis of adipocyte differentiation. Glucose oxidase method was used to detect the effect of OA on glucose consumption in adipocytes. The concentrations of free fatty acids in supernatants were determined by enzyme-linked immunosorbent assay. The expression of PPAR-γ and GLUT-4 mRNA in adipocytes was detected by RT-PCR. Results: Compared with the vehicle control group, OA significantly increased the glucose consumption (P <0.05 or P <0.01), and the effect was significantly enhanced with the increase of the dose in the concentration range of 1 ~ 30 μmol / L. In the range of 3 ~ 30μmol / L, the optical density of oil red O in OA group was significantly higher than that in vehicle control group (P <0.05 or P <0.01). At the same time, OA also reduced the production of free fatty acids. At 30μmol / L, the content of free fatty acids was only 47.2% in the vehicle control group, which was significantly lower than that in the vehicle control group (P <0.01). Compared with the vehicle control group, 30μmol / L OA significantly increased the expression of PPAR-γ and GLUT-4 mRNA in adipocytes (P <0.05). Conclusion: OA can promote adipocyte differentiation and glucose consumption and reduce the production of free fatty acids. The mechanism may be related to up-regulating the expression of PPAR-γ and GLUT-4 mRNA in adipocytes and increasing the sensitivity of adipocytes to insulin.