摘要
目的基于核因子E2相关因子2(nuclear factor erythroid 2-related factor,Nrf2)信号通路观察通腑养髓方对Wilson病(Wilson’s disease,WD)模型TX小鼠神经细胞铁死亡的干预效应,并探讨通腑养髓方对WD神经细胞损伤的保护机制。方法以DL小鼠为正常对照,将TX小鼠随机分为模型组和通腑养髓方组,通腑养髓方组以通腑养髓方中药灌胃治疗30 d,模型组和对照组小鼠以生理盐水灌胃,末次灌胃后取各组小鼠脑组织。采用电感耦合等离子体质谱法检测小鼠脑组织铜、铁微量元素含量,免疫荧光染色技术检测小鼠脑组织转铁蛋白受体(transferrin receptor,TfR)表达水平,比色法检测小鼠脑组织超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehyde,MDA)含量,透射电子显微镜观察小鼠脑组织细胞线粒体形态变化,Fluoro-Jade B(FJB)染色观察小鼠神经细胞损伤程度,Western blot法检测小鼠脑组织铁死亡及Nrf2信号通路相关蛋白[Nrf2,铁蛋白重链(ferritin heavy chain,FTH1)、血红素加氧酶1(heme oxygenase 1,HO1)、醌氧化还原酶1(NADPH quinone oxidoreductase 1,NQO1)]表达水平。结果与正常对照组比较,模型组小鼠脑内铜、铁含量,TfR表达水平,MDA含量显著升高(P<0.05),SOD活性显著下降(P<0.05);与模型组比较,通腑养髓方组小鼠脑内铁含量,TfR表达水平,MDA含量显著降低(P<0.05),SOD活性显著升高(P<0.05)。模型组小鼠脑组织线粒体膜皱缩,嵴数量减少或消失,空泡产生;通腑养髓方组小鼠脑组织线粒体结构损伤程度明显减轻。与正常对照组比较,模型组小鼠神经细胞损伤程度显著增加(P<0.05);与模型组比较,通腑养髓方组神经细胞损伤程度显著减少(P<0.05)。模型组小鼠脑内谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)表达水平较正常对照组显著降低(P<0.05),通腑养髓方组脑内GPX4表达水平较模型组显著升高(P<0.05)。模型组小鼠脑内Nrf2、FTH1、HO1、NQO
Objective To investigate the intervention effect of Tongfu Yangsui prescription on ferroptosis of neural cells in TX mice with Wilson’s disease(WD)based on the nuclear factor erythroid 2-related factor(Nrf2)signaling pathway,as well as the protective mechanism of Tongfu Yangsui prescription against neural cell injury in WD.Methods With DL mice as normal control group,TX mice were randomly divided into model group and Tongfu Yangsui prescription group.The mice in the Tongfu Yangsui prescription group were given the traditional Chinese medicine Tongfu Yangsui prescription by gavage for 30 days,and those in the model group and the normal control group were given normal saline by gavage.Brain tissue samples were collected after the last administration by gavage.Inductively coupled plasma mass spectrometry was used to measure the content of the trace elements copper and iron in brain tissue;immunofluorescent staining was used to measure the expression level of transferrin receptor(TfR)in brain tissue;colorimetry was used to measure the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in brain tissue;transmission electron microscopy was used to observe the morphological changes of mitochondria in brain cells;Fluoro-JadeB(FJB)staining was used to observe the degree of neural cell injury in mice;Western blot was used to measure the expression of the proteins associated with ferroptosis and the Nrf2 signaling pathway in brain tissue,i.e.,Nrf2,ferritin heavy chain(FTH1),heme oxygenase 1(HO1),and NADPH quinone oxidoreductase 1(NQO1).Results Compared with the normal control group,the model group had significant increases in the content of copper and iron,the expression level of TfR,and the content of MDA(P<0.05)and a significant reduction in the activity of SOD in brain tissue(P<0.05);compared with the model group,the Tongfu Yangsui prescription group had significant reductions in the content of iron,the expression level of TfR,and the content of MDA(P<0.05)and a significant increase in the activ
作者
李东昇
程楠
董健健
徐陈陈
耿昊
高曼莉
LI Dong-sheng;CHENG Nan;DONG Jian-jian;XU Chen-chen;GENG Hao;GAO Man-li(Graduate School of Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Affiliated Hospital of Neurology Institute,Anhui University of Chinese Medicine,Anhui Hefei 230061,China)
出处
《安徽中医药大学学报》
CAS
2022年第5期95-101,共7页
Journal of Anhui University of Chinese Medicine
基金
国家自然科学基金项目(81673948,81603596,81904086)。
关键词
WILSON病
铁死亡
氧化应激
Nrf2通路
通腑养髓方
Wilson’s disease
Ferroptosis
Oxidative stress
Nuclear factor erythroid 2-related factor 2 pathway
Tongfu Yangsui prescription
