汉黄芩素通过NRF2/HO-1信号途径诱导大鼠CIA-FLS细胞铁死亡  

Wogonin induces ferroptosis of rat CIA-FLS cells via NRF2/HO-1 signaling pathway

作　　者：何凌飞 张超凡 连杰 申傲轩 董倩男 康晓 吴昊 HE Lingfei;ZHANG Chaofan;LIAN Jie;SHEN Aoxuan;DONG Qiannan;KANG Xiao;WU Hao(Hubei Enshi College,Enshi 455099,China)

机构地区：[1]湖北恩施学院,湖北恩施455099

出　　处：《中国病理生理杂志》2024年第7期1276-1282,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81960776);湖北省自然科学基金资助项目(No.2022CFB515);恩施土家族苗族自治州科技局技术支撑类项目(No.D20230022);湖北恩施学院“两院七所”建设科研项目(No.KYQS202301)。

摘　　要：目的:探讨汉黄芩素(WOG)通过核因子E2相关因子2(NRF2)/血红素加氧酶1(HO-1)信号途径诱导胶原诱导性关节炎大鼠成纤维细胞样滑膜细胞(大鼠CIA-FLS细胞)铁死亡的机制。方法:将大鼠CIA-FLS细胞分为:对照组、低、中、高剂量(25、50和100μmol/L)汉黄芩素组、铁死亡抑制剂(LIP-1)组、LIP-1+高剂量汉黄芩素组、HO-1激动剂钴原卟啉(COPP)组和COPP+高剂量汉黄芩素组,CCK-8法检测细胞活力;结晶紫染色法检测细胞形态;检测氧化应激标志物谷胱甘肽(GSH)、丙二醛(MDA)和超氧化物歧化酶(SOD)的水平;DCFH-DA荧光探针检测细胞内活性氧(ROS)的含量,Western blot检测Kelch样ECH关联蛋白1(KEAP-1)、NRF2和HO-1蛋白表达水平。结果:与正常对照组相比,给予WOG处理后,大鼠CIA-FLS细胞活力显著下降(P<0.01),氧化应激水平显著上升(P<0.01),ROS含量显著增加(P<0.01),NRF2和HO-1蛋白表达水平显著下降(P<0.01),KEAP-1水平显著增加(P<0.01);与WOG组相比,LIP-1处理组的细胞活力显著上升(P<0.01),氧化应激水平显著下降(P<0.01),ROS含量显著减少(P<0.01);与WOG组相比,加入COPP后,NRF2和HO-1蛋白表达水平显著上升(P<0.01),KEAP-1水平显著下降(P<0.01)。结论:WOG能通过NRF2/HO-1信号途径,促进氧化应激来诱导大鼠CIA-FLS细胞铁死亡。AIM:To investigate the mechanism by which wogonin(WOG)induces ferroptosis in collagen-induced arthritis rat fibroblast-like synoviocytes(rat CIA-FLS cells)through the nuclear factor E2-related factor 2(NRF2)/heme oxygenase-1(HO-1)signaling pathway.METHODS:Rat CIA-FLS cells were divided into:control group,low,medium,and high dose of(25,50 and 100μmol/L)WOG group,ferroptosis inhibitor(LIP-1)group,LIP-1+high dose WOG group,HO-1 agonist cobalt protoporphyrin(COPP)group,and COPP+high dose WOG group.CCK-8 assay was used for cell viability.Crystal violet staining was used for for cell morphology.The levels of oxidative stress markers glutathione(GSH),malondialdehyde(MDA),and superoxide dismutase(SOD)were measured.DCFH-DA fluorescent probe was used to detect the intracellular reactive oxygen species(ROS)content as well as Western blot to detect the protein expression levels of Kelch-like ECH-associated protein 1(KEAP-1),NRF2 and HO-1.RESULTS:Compared with the normal control group,administration of WOG treatment resulted in a significant decrease in CIA-FLS cell viability(P<0.01),a significant increase in the level of oxidative stress(P<0.01),a significant increase in the content of ROS(P<0.01),a significant decrease in the level of expression of NRF2 and HO-1 proteins(P<0.01),and a significant increase in the level of KEAP-1(P<0.01)in the rat.Compared with the WOG group,the LIP-1-treated group showed a significant increase in cell viability(P<0.01),a significant decrease in the level of oxidative stress(P<0.01),and a significant decrease in the content of ROS(P<0.01).Compared with the WOG group,the addition of COPP resulted in a significant increase in the protein expression levels of NRF2 and HO-1(P<0.01)and a significant decrease in KEAP-1 levels(P<0.01).CONCLUSION:WOG can induce ferroptosis in rat CIA-FLS cells by promoting oxidative stress through the NRF2/HO-1 signaling pathway.

关 键 词：汉黄芩素 类风湿关节炎 CIA-FLS细胞 NRF2/HO-1信号通路 铁死亡 

分 类 号：R593.21[医药卫生—内科学] R363.2[医药卫生—临床医学] R285.5