摘要
目的:探讨通窍明目汤改善青光眼视神经损伤的分子机制。方法:通过网络药理学方法,数据库检索药物、疾病及铁死亡数据库靶点,筛选出共同靶点与关键靶标,构建“通窍明目汤-青光眼-铁死亡”网络,并进行通路富集分析;体外实验以过氧化氢诱导RGC,造模成功后,随机分为模型组、通窍明目汤组、阳性药物组,共3组,另设正常空白组,自第5周开始予通窍明目汤(99.96 g/kg)灌胃干预。实验结束后,CCK-8检测RGC增殖,流式细胞术检测ROS表达,ELISA法检测RGC中铁离子、MDA、SOD、GSH-PX含量及活性,Western blot检测RGC中ACSL4、GPX-4、FTH-1蛋白及p53/SLC7A11相关蛋白的表达变化,RT-PCR法检测RGC中p53、SLC7A11 mRNA含量。结果:经通窍明目汤汤干预后,与模型组比较,通窍明目汤显著降低RGC内活性氧阳性率、铁离子水平及MDA含量、下调RGC细胞核内p53、ACSL4及ACSL4、p53 mRNA表达,增加RGC增殖率,升高SOD、CAT及GSH水平,上调GPX-4、FTH-1、SLC7A11及GPX-4、FTH-1、SLC7A11 mRNA表达(P<0.05)。结论:通窍明目汤可能通过多成分、多靶点,涉及多条分子通路,其机制可能与阻断p53/SLC7A11信号通路中细胞内p53的表达及systemXc-所介导的RGC铁死亡,抑制SLC7A11表达,导致胱氨酸减少,增加铁死亡敏感性,触发的氧化应激和过度炎症反应,从而避免细胞受损和功能紊乱有关。
Objective:To explore the molecular mechanism of Tongqiao Mingmu Decoction in improving optic nerve damage in glaucoma.Method:Using network pharmacology methods,drug,disease,and ferroptosis database targets were searched,common and key targets were screened,and a"Tongqiao Mingmu Decoction-glaucoma-ferroptosis"network was constructed,followed by pathway enrichment analysis;In vitro experiments induced RGC with hydrogen peroxide.After successful modeling,the mice were randomly divided into three groups:model group,Tongqiao Mingmu Decoction group,and positive drug group.A normal blank group was also set up,and Tongqiao Mingmu Decoction(99.96 g/kg)was administered by gavage intervention starting from week 5.After the experiment,CCK-8 was used to detect RGC proliferation,and flow cytometry was used to detect ROS expression.ELISA was used to detect the content and activity of iron ions,MDA,SOD,GSH-PX in RGC.Western blot was used to detect the expression changes of ACSL4,GPX-4,FTH-1 proteins and p53/SLC7A11 related proteins in RGC,and RT-PCR was used to detect the mRNA content of p53 and SLC7A11 in RGC.Result:After intervention with Tongqiao Mingmu Decoction,compared with the model group,Tongqiao Mingmu Decoction significantly reduced the positive rate of reactive oxygen species,iron ion levels,and MDA content in RGC,downregulated the expression of p53,ACSL4,ACSL4,and p53 mRNA in RGC nucleus,increased RGC proliferation rate,increased SOD,CAT,and GSH levels,and upregulated the expression of GPX-4,FTH-1,SLC7A11,GPX-4,FTH-1,and SLC7A11 mRNA(P<0.05).Conclusion:Tongqiao Mingmu Decoction may involve multiple molecular pathways through multiple components and targets,and its mechanism may be related to blocking the expression of intracellular p53 in the p53/SLC7A11 signaling pathway and the RGC ferroptosis mediated by system Xc-,inhibiting the expression of SLC7A11,leading to a decrease in cysteine,increasing ferroptosis sensitivity,triggering oxidative stress and excessive inflammatory response,thereby avoiding cell damage and
作者
杨稀瑞
王继雪
王思琦
孙会兰
袁星星
孙河
YANG Xirui;WANG Jixue;WANG Siqi;SUN Huilan;YUAN Xingxing;SUN He(Ophthalmology Department of the First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou,Henan 450000,China;Peripheral Vascular Department of the First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou,Henan 450000,China;The First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou,Henan 450000,China;Heilongjiang University of Traditional Chinese Medicine,Harbin,Heilongjiang 150001,China;The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin,Heilongjiang 150040,China)
出处
《海南医学院学报》
CAS
北大核心
2024年第22期1717-1726,共10页
Journal of Hainan Medical University
基金
国家自然科学基金青年基金项目(82205195,82205117)
河南省中医临床研究基地科研专项(2022JDZX127,2022JDZX135)
河南省医学科技攻关计划项目(LHGJ20230686)。
