目的 观察S-雌马酚(S-equol,S-Eq)对油酸钠(sodium oleate,NaOL)诱导的BRL细胞脂肪变性的影响,并探讨其作用机制。方法 体外培养BRL细胞,以不同浓度的S-Eq、NaOL、PHTPP处理48 h,通过CCK-8法检测细胞存活率、油红O染色及细胞TG检测确定诱导BRL细胞脂肪病变模型的NaOL以及S-Eq、PHTPP干预的适宜浓度。将细胞分为对照组、模型组,模型+S-Eq低、高浓度组(10-6 mol/L,10-5 mol/L),模型+雌二醇干预组(estradiol,E2, 10-7 mol/L),模型+ S-Eq(10-5 mol/L)+PHTPP (10-6 mol/L)。干预48 h后检测细胞TG、TNF-α、IL-6含量,qRT-PCR检测细胞TNF-α、IL-6 mRNA表达水平,Western bloting检测细胞ERβ,PI3k ,AKT,p-AKT,p-p65蛋白表达水平。结果 与对照组比较,当NaOL浓度为0.48 mmol/L时BRL细胞活力无明显减低、TG含量明显增加,同时细胞出现大量脂滴聚集,确定为诱导细胞脂肪病变模型的适宜浓度;干预结束时,与模型组比较,S-Eq或E2干预可显著减少细胞TG、TNF-α及IL-6水平(P<0.05),同时ERβ、PI3k 、p-AKT表达水平显著升高(P<0.05),p-p65、TNF-α、IL-6表达水平显著降低(P<0.05);PHTPP可显著抑制S-Eq对细胞脂肪变性的改善作用。结论 S-Eq可有效改善NaOL诱导的BRL细胞脂肪变性,其部分机制与S-Eq通过ERβ调控PI3K/AKT/NF-κB信号通路减轻炎症反应有关。
Abstract
Objective To observe the effect of S-equol (S-Eq) on sodium oleate (NaOL)-induced steatosis in BRL cells, and to investigate its mechanism of action. Methods BRL cells were cultured in vitro, treated with different concentrations of S-Eq, NaOL and PHTPP for 48h. The appropriate concentration of NaOL for the induction of BRL cell steatosis and the appropriate concentration of S-Eq and PHTPP for intervention were determined by cell viability, oil red O staining and cell TG content. The cells were divided into control group, model group, model + low and high S-Eq concentration (10-6 mol/L, 10-5 mol/L) groups, model + estradiol (E2, 10-7 mol/L) group, model + S-Eq (10-5 mol/L) + Fulvestrant (10-6 mol/L) group. After 48 h of intervention, the contents of TG, TNF-α and IL-6 were detected in cells, the expression levels of TNF-α and IL-6 mRNA were detected by qRT-PCR, and the expression levels of ERβ, PI3k, AKT, p-AKT, p-p-p65 proteins were detected by Western blotting. Results Compared with the control group, when the NaOL concentration was 0.48 mmol/L, the viability of BRL cells did not decrease significantly, the content of TG was significantly increased, and a large number of lipid droplets were accumulated in the cells, which was determined as the appropriate concentration of NaOL for inducing steatosis. At the end of the intervention, compared with the model group, S-Eq or E2 intervention could significantly reduce the contents of TG, TNF-α and IL-6 (P<0.05), as well as the expression levels of TNF-α and IL-6 mRNA in cells (P<0.05). Meanwhile, the expressions of ERβ, PI3k, p-AKT were significant increased, and the expression level of p-p65 protein was significanty reduced (P<0.05). PHTPP could significantly inhibit the improvement effect of S-Eq on cell steatosis. Conclusion S-Eq could effectively improve NAOL-induced BRL cell steatosis, and part of the mechanism is related to the reduction of inflammatory response by S-Eq via regulating the PI3K/AKT/NF-κB signaling pathway through ERβ.
关键词
S-雌马酚 /
脂肪变性 /
磷脂酰肌醇3-激酶/丝氨酸苏氨酸蛋白激酶 /
核因子κB
Key words
s-equol /
steatosis /
phosphatidylinositol 3-kinase/serine-threonine protein kinase /
nuclear factor kappa-B
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基金
国家自然基金面上项目（No.81973040）
