S-雌马酚通过ERβ调控PI3K/AKT/NF-κB信号通路改善油酸钠诱导的BRL细胞脂肪变性

Add to Favorite S-EQUOL IMPROVES SODIUM OLEATE-INDUCED STEATOSIS BY REGULATING PI3K/AKT/NF-κB SIGNALING PATHWAY THROUGH ERβIN BRL CELLS

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,E210^(-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。

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-αand IL-6 were detected in cells,the expression levels of TNF-αand 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 E2intervention could significantly reduce the contents of TG,TNF-αand IL-6(P<0.05),as well as the expression levels of TNF-αand 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.