丹参酮IIA抑制肠上皮细胞铁死亡的作用与机制研究  

The inhibitory effect of tanshinone IIA on intestinal epithelial cell ferroptosis and its mechanisms

作　　者：刘翔[1,3] 孙扬[2] 马丽莉 马颖才[4] LIU Xiang;SUN Yang;MA Li-li;MA Ying-cai(Suzhou Medical College of Soochow University,Jiangsu 215000,China;School of Basic Medicine,Qingdao University,Shandong 266071,China;Department of Emergency Internal Medicine,Qinghai Provincial People's Hospital,Xining 810000,China;Department of Gastroenterology,Qinghai Provincial People's Hospital,Xining 810000,China)

机构地区：[1]苏州大学苏州医学院,江苏215000 [2]青岛大学基础医学院,山东266071 [3]青海省人民医院急诊内科,西宁810000 [4]青海省人民医院消化内科,西宁810000

出　　处：《中国医药生物技术》2023年第6期550-557,共8页Chinese Medicinal Biotechnology

摘　　要：目的阐释丹参酮IIA对肠上皮细胞铁死亡的作用及其调控机制。方法利用铁死亡诱导剂RSL3处理大鼠IEC-6细胞建立肠上皮细胞铁死亡模型;利用CCK8方法测定细胞活性;利用姬姆萨染色和透射电镜技术观察细胞形态学和结构;利用超高效液相色谱分离和质谱分析进行非靶向代谢物检测;利用RT-qPCR检测相关基因mRNA水平。结果RSL3能够显著诱导肠上皮IEC-6细胞的死亡,并可被铁死亡的抑制剂Fer-1逆转;IEC-6细胞铁死亡形态学结构特征包括线粒体变小、嵴减少、膜密度增高及外膜断裂、ROS聚集等;丹参酮IIA能够逆转RSL3诱导的细胞铁死亡;丹参酮IIA表现出抑制IEC-6细胞铁死亡的活性;非靶向代谢组学数据表明丹参酮IIA能提高细胞内谷胱甘肽的含量,并上调GPX4基因表达。结论丹参酮IIA能够有效抑制肠上皮细胞的铁死亡;丹参酮IIA影响谷胱甘肽的代谢和上调GPX4表达。靶向上皮细胞铁死亡可能是丹参酮IIA治疗炎症性肠病的重要机制。Objective Aim to clarify the effect of tanshinone IIA on ferroptosis of intestinal epithelial cells,as well as its mechanisms.Methods The ferroptosis model of intestinal epithelial cells was established using the RSL3-treated rat intestinal cell No.6(IEC-6)cells.Cell viability was determined by CCK8 assay.Cellular morphology and structure were characterized by Giemsa staining and transmission electron microscopy techniques;Untargeted metabolites were detected using ultra-performance liquid chromatography separation and mass spectrometry analysis.RT-qPCR was used to detect the mRNA levels of relevant genes.Results We demonstrated that RSL3-induced IEC-6 cell death is an ideal ferroptosis model of epithelial cells.RSL3 significantly induced ferroptosis of intestinal epithelial IEC-6 cells,which could be reversed by a ferroptosis inhibitor Fer-1.The structural features of ferroptosis in IEC-6 cells included mitochondrial shrinkage,increased membrane density and membrane rupture,and ROS aggregation,etc.Tanshinone IIA treatment reversed RSL3-induced cell death and exhibited inhibitory activity against ferroptosis in IEC-6 cells.Untargeted metabolomic data showed that tanshinone IIA increased the concentration of glutathione and upregulated GPX4 gene expression.Conclusions Tanshinone IIA could effectively reverse the ferroptosis of intestinal epithelial cells by affecting the metabolism of glutathione and increasing GPX4 expression.Targeting ferroptosis of intestinal epithelial cells may be an important mechanism for the therapeutic effect of tanshinone IIA on IBD.

关 键 词：炎症性肠病 铁死亡 丹参酮IIA 谷胱甘肽 GPX4 

分 类 号：R285[医药卫生—中药学]