山药中熊果苷通过ERβ抑制LPS诱导NRK-52e细胞凋亡

Arbutin isolated from Chinese yam inhibits LPS-induced NRK-52e cell apoptosis by ERβ

目的探究山药中熊果苷(arbutin,Ar)对LPS诱导NRK-52e细胞凋亡的干预作用及潜在机制。方法建立LPS诱导的NRK-52e细胞损伤模型,将细胞分为正常组、LPS组(1 mg·L^(-1))、低剂量Ar(LPS,1 mg·L^(-1)+Ar,5μmol·L^(-1))和高剂量Ar(LPS,1 mg·L^(-1)+Ar,10μmol·L^(-1))及其对应的抑制剂THC组(1μmol·L^(-1)),检测细胞活力;流式细胞术检测ROS、细胞凋亡水平、Ca2+浓度及线粒体膜电位;In cell western技术检测细胞凋亡关键蛋白表达水平;结果Ar可有效调节LPS诱导的NRK-52e细胞ROS水平、Ca^(2+)浓度、凋亡关键蛋白和ERβ水平,抑制线粒体膜电位下降,但是该作用被雌激素受体β抑制剂THC所阻断,且Ar与ERβ具有较好的结合活性。结论Ar可能通过ERβ抑制LPS诱导NRK-52e细胞凋亡。

Aim To investigate the effect of arbutin on apoptosis of NRK-52e cells induced by LPS and the potential mechanism.Methods The model of NRK-52e cells injury was constructed by LPS,and NRK-52e cells were divided into control,LPS(1 mg·L^(-1)),low dose arbutin(LPS,1 mg·L^(-1)+arbutin,5μmol·L^(-1)),high dose arbutin(LPS,1 mg·L^(-1)+arbutin,10μmol·L^(-1))and its corresponding inhibitor THC group(1μmol·L^(-1)).The cell viability was detected;the levels of ROS,apoptosis,Ca2+concentration and mitochondrial membrane potential(MMP)were detected by flow cytometry;the levels of key apoptosis proteins were detected by in cell western;the binding activity of arbutin with ERβwas imitated by molecular docking technology,and verified by in cell western.Results Arbutin could effectively regulate the levels of ROS,Ca^(2+),apoptosis proteins and ERβin NRK-52e cells induced by LPS and inhibit the decline of MMP,which is blocked by estrogen receptorβinhibitor THC.In addition,arbutin has good binding activity with ERβ.Conclusion This study confirms that arbutin could inhibit LPS-induced apoptosis of NRK-52e cells through ERβ.