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目的:观察不同质量浓度熊果酸(UA)对RAW264.7巨噬细胞源性泡沫细胞促进胆固醇流出的影响及过氧化物酶体增殖物激活受体-γ(PPAR-γ)转运蛋白的表达,探讨可能作用机制。方法:体外培养RAW264.7巨噬细胞,用20 mg·L~(-1)的氧化型低密度脂蛋白(ox-LDL)孵育48 h诱导成泡沫细胞,采用油红O染色,光镜下鉴定泡沫细胞形态变化,液体闪烁计数仪检测胆固醇的流出率,并用实时荧光定量聚合酶链式反应(Real-time PCR)及酶联免疫吸附(ELISA)测定PPAR-γ的基因及蛋白表达。结果:巨噬细胞经ox-LDL诱48 h后转化为泡沫细胞,与空白组比较,10,15,20,25 mg·L~(-1)质量浓度UA干预组泡沫细胞的胆固醇流出率上升(P<0.01);10,15,20,25 mg·L~(-1)质量浓度UA干预组泡沫细胞内PPAR-γ的基因表达上调(P<0.01),在一定质量浓度范围内呈剂量依赖性;10,15,20,25 mg·L~(-1)质量浓度UA干预组泡沫细胞内PPAR-γ的蛋白表达增加(P<0.01),差异有统计学意义。结论:经20 mg·L~(-1)ox-LDL的诱导后,巨噬细胞分化为泡沫细胞,细胞内脂质大量增加,UA促进巨噬细胞内胆固醇流出,可能与上调细胞内PPAR-γ的表达有关。
OBJECTIVE: To investigate the effects of different concentrations of UA on the cholesterol efflux in RAW264.7 macrophage-derived foam cells and the expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) transporter , To explore the possible mechanism. Methods: RAW264.7 macrophages were cultured in vitro and were induced to foam cells by ox-LDL (20 mg · L -1) for 48 h. The cells were stained with oil red O, The morphological changes of foam cells were identified. The efflux rate of cholesterol was detected by liquid scintillation counter. The gene and protein expression of PPAR-γ were detected by Real-time PCR and ELISA. RESULTS: After induced by ox-LDL for 48 h, macrophages were transformed into foam cells. Compared with the blank group, the rate of cholesterol efflux from foam cells was increased in the 10, 15, 20 and 25 mg · L -1 concentrations of UA (P <0.01). The gene expression of PPAR-γ was up-regulated in foam cells treated with 10, 15, 20 and 25 mg · L -1 UA (P <0.01) Dependent. The protein expression of PPAR-γ in foam cells increased with 10, 15, 20, 25 mg · L -1 concentration of UA (P <0.01), and the difference was statistically significant. CONCLUSION: Macrophages differentiate into foam cells induced by 20 mg · L -1 ox-LDL, the intracellular lipid is greatly increased, and UA promotes the efflux of cholesterol from macrophages, which may be related to upregulation of intracellular PPAR- γ expression.