Article(id=1276976095077462705, tenantId=1146029695717560320, journalId=1276577828834349082, issueId=1276975955985961244, articleNumber=null, orderNo=null, doi=10.19540/j.cnki.cjcmm.20250825.501, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1749052800000, receivedDateStr=2025-06-05, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1782384426135, onlineDateStr=2026-06-25, pubDate=1767196800000, pubDateStr=2026-01-01, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782384426135, onlineIssueDateStr=2026-06-25, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782384426135, creator=13701087609, updateTime=1782384426135, updator=13701087609, issue=Issue{id=1276975955985961244, tenantId=1146029695717560320, journalId=1276577828834349082, year='2026', volume='51', issue='1', pageStart='1', pageEnd='300', issueExtLink='null', onlineDate='null', pubDate='1767196800000', pubDateStr='2026-01-01', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782384392973, creator='13701087609', updateTime=1782384484405, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276976339546673871, tenantId=1146029695717560320, journalId=1276577828834349082, issueId=1276975955985961244, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276976339546673872, tenantId=1146029695717560320, journalId=1276577828834349082, issueId=1276975955985961244, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=269, endPage=280, ext={EN=ArticleExt(id=1276976095308149427, articleId=1276976095077462705, tenantId=1146029695717560320, journalId=1276577828834349082, language=EN, title=Xihuang Pills induce mitochondria-associated ferroptosis to enhance therapeutic efficacy of temozolomide against glioblastoma via Nrf2/HO-1/GPX4 signaling axis, columnId=null, journalTitle=Chinese Journal of Traditional Chinese Medicine, columnName=null, runingTitle=null, highlight=null, articleAbstract=

This study aimed to investigate whether Xihuang Pills (XHP) enhance the therapeutic efficacy of temozolomide (TMZ) against glioblastoma (GBM) U251 cells by inducing mitochondria-associated ferroptosis via regulation of Nrf2/HO-1/GPX4 signaling axis. The XHP-containing serum was prepared, and the effects of various concentrations of TMZ (0-200 μmol·L-1) and XHP (0-20%) on U251 cell viability were assessed by CCK-8 assay. The optimal synergistic dose (200 μmol·L-1 TMZ + 10% XHP) was identified via the SynergyFinder platform for establishing a combined intervention model. Control, XHP, TMZ, and XHP + TMZ groups were designed. Cell proliferation was evaluated by EdU staining and colony formation assay, and cell migration and invasion were assessed by Transwell and wound healing assays. FerroOrange and DCFH-DA fluorescent probes were used to measure the intracellular levels of Fe2+ and ROS, respectively. The JC-1 fluorescent probe was used to assess mitochondrial membrane potential, while Mito-PeDPP fluorescent probe to examine mitochondrial lipid peroxidation. Mito-Tracker Green and MitoSOXTM Red were used to examine mitochondrial morphology and oxidative stress. Transmission electron microscopy was adopted to observe the mitochondrial ultrastructure. GSH and MDA levels were measured by colorimetric assays. Western blot was employed to assess the protein levels of Nrf2, HO-1, xCT, and GPX4. The results showed that, XHP + TMZ suppressed cell viability, proliferation, migration, and invasion (P<0.01), increased Fe2+ accumulation, ROS generation, GSH depletion, and lipid peroxidation (P<0.001), reduced mitochondrial membrane potential, caused severe structural damage, and down-regulated the protein levels of Nrf2, HO-1, GPX4, and xCT, which indicated that the antioxidant ferroptosis-protective pathway was co-inhibited. In conclusion, XHP enhances TMZ-induced ferroptosis in GBM cells by suppressing the Nrf2/HO-1/GPX4 signaling axis. The findings provide new mechanism insights into the synergistic antitumor effect of traditional Chinese medicine in glioblastoma therapy and offer experimental evidence for the treatment of refractory brain tumors with combined strategies.

, authors=null, authorsList=Yuan-yuan ZHU, Xue-ling ZHAO, Nuo-jin GENG, Xin-hua ZHU, Ning TANG, authorCompany=null, correspAuthors=Ning TANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1276976095564001973, articleId=1276976095077462705, tenantId=1146029695717560320, journalId=1276577828834349082, language=CN, title=基于Nrf2/HO-1/GPX4信号轴探究西黄丸诱导线粒体相关性铁死亡增敏替莫唑胺治疗胶质母细胞瘤的机制, columnId=1276975994707775974, journalTitle=中国中药杂志, columnName=药理, runingTitle=null, highlight=null, articleAbstract=

探讨西黄丸(Xihuang Pills, XHP)通过调控核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)/谷胱甘肽过氧化物酶4(GPX4)信号轴,诱导线粒体相关性铁死亡,从而增敏替莫唑胺(TMZ)对胶质母细胞瘤(GBM)U251细胞的治疗效应。实验制备XHP含药血清,采用CCK-8法评估不同浓度TMZ(0~200 μmol·L-1)和XHP(0~20%)对U251细胞活力的影响,通过SynergyFinder平台筛选最佳联合抑制剂量(200 μmol·L-1 TMZ+10% XHP),建立联合干预模型。设置对照组、XHP组、TMZ组、XHP+TMZ组,采用EdU染色与克隆形成实验评估细胞增殖能力,Transwell小室与划痕实验检测侵袭迁移;FerroOrange荧光探针检测Fe2+水平,DCFH-DA荧光探针检测活性氧(ROS)水平,JC-1荧光探针检测线粒体膜电位,Mito-PeDPP荧光探针检测线粒体内脂质过氧化水平,Mito-Tracker Green和Mito-SOXTM Red荧光染料检测线粒体形态结构和过氧化水平,透射电镜观察线粒体超微结构,比色法测定谷胱甘肽(GSH)与丙二醛(MDA)水平,蛋白免疫印迹法(Western blot)检测Nrf2、HO-1、溶质载体家族成员SLC7A11(xCT)、GPX4蛋白表达。结果显示,XHP+TMZ联合处理显著抑制细胞活力、增殖及迁移侵袭能力(P<0.01),并加重Fe2+积累、ROS生成、GSH耗竭及脂质过氧化(P<0.001);线粒体膜电位明显丧失,结构严重破坏;Nrf2、HO-1、GPX4、xCT蛋白表达明显下调,提示抗氧化铁死亡通路被协同抑制。结果表明,XHP通过抑制Nrf2/HO-1/GPX4信号轴,诱导线粒体相关性铁死亡,从而显著增强TMZ对GBM细胞的治疗效果,揭示中医药协同抗脑胶质瘤的新机制,为难治性脑瘤的联合治疗提供了实验依据与理论支撑。

, authors=

祝圆圆,住院医师,主要从事中西结合防治脑胶质瘤研究,E-mail:

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*

唐宁,主治医师,主要从事中西结合防治脑胶质瘤研究,E-mail:
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祝圆圆,住院医师,主要从事中西结合防治脑胶质瘤研究,E-mail:

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基于Nrf2/HO-1/GPX4信号轴探究西黄丸诱导线粒体相关性铁死亡增敏替莫唑胺治疗胶质母细胞瘤的机制
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祝圆圆 , 赵雪玲 , 耿诺金 , 朱新华 , 唐宁 *
中国中药杂志 | 药理 2026,51(1): 269-280
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中国中药杂志 |药理 2026 , 51 (1) : 269 -280
基于Nrf2/HO-1/GPX4信号轴探究西黄丸诱导线粒体相关性铁死亡增敏替莫唑胺治疗胶质母细胞瘤的机制
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祝圆圆 , 赵雪玲, 耿诺金, 朱新华, 唐宁*
作者信息
  • 湖南中医药大学 第一附属医院,湖南 长沙 410007
通讯作者:

*

唐宁,主治医师,主要从事中西结合防治脑胶质瘤研究,E-mail:
Xihuang Pills induce mitochondria-associated ferroptosis to enhance therapeutic efficacy of temozolomide against glioblastoma via Nrf2/HO-1/GPX4 signaling axis
Yuan-yuan ZHU , Xue-ling ZHAO, Nuo-jin GENG, Xin-hua ZHU, Ning TANG*
Affiliations
  • the First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China
出版时间: 2026-01-01 doi: 10.19540/j.cnki.cjcmm.20250825.501
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探讨西黄丸(Xihuang Pills, XHP)通过调控核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)/谷胱甘肽过氧化物酶4(GPX4)信号轴,诱导线粒体相关性铁死亡,从而增敏替莫唑胺(TMZ)对胶质母细胞瘤(GBM)U251细胞的治疗效应。实验制备XHP含药血清,采用CCK-8法评估不同浓度TMZ(0~200 μmol·L-1)和XHP(0~20%)对U251细胞活力的影响,通过SynergyFinder平台筛选最佳联合抑制剂量(200 μmol·L-1 TMZ+10% XHP),建立联合干预模型。设置对照组、XHP组、TMZ组、XHP+TMZ组,采用EdU染色与克隆形成实验评估细胞增殖能力,Transwell小室与划痕实验检测侵袭迁移;FerroOrange荧光探针检测Fe2+水平,DCFH-DA荧光探针检测活性氧(ROS)水平,JC-1荧光探针检测线粒体膜电位,Mito-PeDPP荧光探针检测线粒体内脂质过氧化水平,Mito-Tracker Green和Mito-SOXTM Red荧光染料检测线粒体形态结构和过氧化水平,透射电镜观察线粒体超微结构,比色法测定谷胱甘肽(GSH)与丙二醛(MDA)水平,蛋白免疫印迹法(Western blot)检测Nrf2、HO-1、溶质载体家族成员SLC7A11(xCT)、GPX4蛋白表达。结果显示,XHP+TMZ联合处理显著抑制细胞活力、增殖及迁移侵袭能力(P<0.01),并加重Fe2+积累、ROS生成、GSH耗竭及脂质过氧化(P<0.001);线粒体膜电位明显丧失,结构严重破坏;Nrf2、HO-1、GPX4、xCT蛋白表达明显下调,提示抗氧化铁死亡通路被协同抑制。结果表明,XHP通过抑制Nrf2/HO-1/GPX4信号轴,诱导线粒体相关性铁死亡,从而显著增强TMZ对GBM细胞的治疗效果,揭示中医药协同抗脑胶质瘤的新机制,为难治性脑瘤的联合治疗提供了实验依据与理论支撑。

西黄丸  /  胶质母细胞瘤  /  替莫唑胺  /  铁死亡  /  线粒体损伤

This study aimed to investigate whether Xihuang Pills (XHP) enhance the therapeutic efficacy of temozolomide (TMZ) against glioblastoma (GBM) U251 cells by inducing mitochondria-associated ferroptosis via regulation of Nrf2/HO-1/GPX4 signaling axis. The XHP-containing serum was prepared, and the effects of various concentrations of TMZ (0-200 μmol·L-1) and XHP (0-20%) on U251 cell viability were assessed by CCK-8 assay. The optimal synergistic dose (200 μmol·L-1 TMZ + 10% XHP) was identified via the SynergyFinder platform for establishing a combined intervention model. Control, XHP, TMZ, and XHP + TMZ groups were designed. Cell proliferation was evaluated by EdU staining and colony formation assay, and cell migration and invasion were assessed by Transwell and wound healing assays. FerroOrange and DCFH-DA fluorescent probes were used to measure the intracellular levels of Fe2+ and ROS, respectively. The JC-1 fluorescent probe was used to assess mitochondrial membrane potential, while Mito-PeDPP fluorescent probe to examine mitochondrial lipid peroxidation. Mito-Tracker Green and MitoSOXTM Red were used to examine mitochondrial morphology and oxidative stress. Transmission electron microscopy was adopted to observe the mitochondrial ultrastructure. GSH and MDA levels were measured by colorimetric assays. Western blot was employed to assess the protein levels of Nrf2, HO-1, xCT, and GPX4. The results showed that, XHP + TMZ suppressed cell viability, proliferation, migration, and invasion (P<0.01), increased Fe2+ accumulation, ROS generation, GSH depletion, and lipid peroxidation (P<0.001), reduced mitochondrial membrane potential, caused severe structural damage, and down-regulated the protein levels of Nrf2, HO-1, GPX4, and xCT, which indicated that the antioxidant ferroptosis-protective pathway was co-inhibited. In conclusion, XHP enhances TMZ-induced ferroptosis in GBM cells by suppressing the Nrf2/HO-1/GPX4 signaling axis. The findings provide new mechanism insights into the synergistic antitumor effect of traditional Chinese medicine in glioblastoma therapy and offer experimental evidence for the treatment of refractory brain tumors with combined strategies.

Xihuang Pills  /  glioblastoma  /  temozolomide  /  ferroptosis  /  mitochondrial damage
祝圆圆, 赵雪玲, 耿诺金, 朱新华, 唐宁. 基于Nrf2/HO-1/GPX4信号轴探究西黄丸诱导线粒体相关性铁死亡增敏替莫唑胺治疗胶质母细胞瘤的机制. 中国中药杂志, 2026 , 51 (1) : 269 -280 . DOI: 10.19540/j.cnki.cjcmm.20250825.501
Yuan-yuan ZHU, Xue-ling ZHAO, Nuo-jin GENG, Xin-hua ZHU, Ning TANG. Xihuang Pills induce mitochondria-associated ferroptosis to enhance therapeutic efficacy of temozolomide against glioblastoma via Nrf2/HO-1/GPX4 signaling axis[J]. Chinese Journal of Traditional Chinese Medicine, 2026 , 51 (1) : 269 -280 . DOI: 10.19540/j.cnki.cjcmm.20250825.501
  • 湖南省卫健委科研计划项目(D202304089721; W20243137)
  • 湖南中医药大学校院联合基金重点项目(2024XYLH005)
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2026年第51卷第1期
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doi: 10.19540/j.cnki.cjcmm.20250825.501
  • 接收时间:2025-06-05
  • 首发时间:2026-06-25
  • 出版时间:2026-01-01
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  • 收稿日期:2025-06-05
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湖南省卫健委科研计划项目(D202304089721; W20243137)
湖南中医药大学校院联合基金重点项目(2024XYLH005)
作者信息
    湖南中医药大学 第一附属医院,湖南 长沙 410007

通讯作者:

*

唐宁,主治医师,主要从事中西结合防治脑胶质瘤研究,E-mail:
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https://castjournals.cast.org.cn/joweb/zgzyzz/CN/10.19540/j.cnki.cjcmm.20250825.501
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2种不同金属材料的力学参数

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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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