加味七方胃痛颗粒含药血清对胃黏膜上皮细胞铁死亡的影响及机制研究

Effects and Mechanism of Modified Qifang Weitong Granules Drug Serum on Ferroptosis of Gastric Epithelial Cells

目的:探究加味七方胃痛颗粒含药血清对胃黏膜上皮细胞铁死亡的影响及机制。方法:GES-1细胞分为空白对照组、模型组、加味七方胃痛颗粒含药血清低浓度组、加味七方胃痛颗粒含药血清中浓度组、加味七方胃痛颗粒含药血清高浓度组、si-E-cad-NC+加味七方胃痛颗粒含药血清组、si-E-cad+加味七方胃痛颗粒含药血清组。CCK-8法检测细胞活力;流式细胞术检测细胞凋亡、ROS水平、线粒体膜电位和Fe~(2+)水平;qPCR检测E-cadherin、NF2、YAP、TEAD、TfR1 mRNA表达;Western Blot检测E-cadherin、Merlin、p-YAP、YAP、TEAD、TfR1蛋白表达。结果:与空白对照组比较,模型组GES-1细胞活力及E-cadherin、NF2(Merlin)、TEAD表达显著降低,细胞凋亡率、ROS水平、线粒体膜电位、Fe~(2+)水平、TfR1 mRNA及蛋白表达和p-YAP/YAP水平显著升高。与模型组比较,加味七方胃痛颗粒含药血清可逆转上述除TEAD外各指标的变化。给予加味七方胃痛颗粒的同时抑制E-cadherin表达则可抑制加味七方胃痛颗粒含药血清的作用。结论:加味七方胃痛颗粒可通过上调E-cadherin表达,活化NF2而拮抗Hippo通路,抑制胃黏膜上皮细胞铁死亡。

Objective:To explore the effect and mechanism of modified Qifang weitong granules drug serum on ferroptosis of gastric epithelial cells.Methods:GES-1 cells were divided into black control group,model group,modified Qifang weitong granules drug serum low,middle,high doses groups,si-E-cad-NC with modified Qifang weitong granules drug serum group,si-E-cad with modified Qifang weitong granules drug serum group.Cell viability was measured by CCK-8;The apoptosis,ROS level,mitochondrial membrane potential and Fe2+level were measured by flow cytometry;The mRNA expressions of E-cadherin,NF2,YAP,TEAD and TfR1 were measured by qPCR,the protein expressions of E-cadherin,Merlin,p-YAP,YAP,TEAD,TfR1 were measured by Western Blot.Results:Compared with blank control group,GES-1 cell viability and expressions of E-cadherin,NF2(Merlin)and TEAD were significantly decreased in model group,while apoptosis rate,ROS level,mitochondrial membrane potential,Fe2+level,TfR1 mRNA and protein expressions and p-YAP/YAP level were significantly increased.Compared with model group,modified Qifang weitong granules drug serum could reverse the changes of the above indexes except TEAD.The effect of modified Qifang weitong granules drug serum was inhibited when modified Qifang weitong granules were given and the expression of E-cadherin was inhibited.Conclusion:Modified Qifang weitong granules can antagonize Hippo pathway and inhibit iron death in gastric mucosal epithelial cells by up-regulating expression of E-cadherin and activating NF2.