Article(id=1238813310288711956, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1238813307784712441, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250765, pmid=null, cstr=null, oa=null, hot=1, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1760284800000, receivedDateStr=2025-10-13, revisedDate=null, revisedDateStr=null, acceptedDate=1765814400000, acceptedDateStr=2025-12-16, onlineDate=1773285709210, onlineDateStr=2026-03-12, pubDate=1772553600000, pubDateStr=2026-03-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773285709210, onlineIssueDateStr=2026-03-12, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773285709210, creator=13701087609, updateTime=1775797808660, updator=13701087609, issue=Issue{id=1238813307784712441, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='3', pageStart='961', pageEnd='1466', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773285708614, creator=13701087609, updateTime=1773291912509, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1238839328915378858, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1238813307784712441, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1238839328915378859, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1238813307784712441, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1225, endPage=1235, ext={EN=ArticleExt(id=1238813310674587941, articleId=1238813310288711956, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Upregulation of Kelch-like ECH-associated protein 1 represses the replication of herpes simplex virus type 1, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the effect of Kelch-like ECH-associated protein 1 (KEAP1) on the replication of herpes simplex virus type 1 (HSV-1) and thus provide theoretical support for anti-herpes simplex virus research. Methods The mRNA and protein levels of molecules in the KEAP1-NRF2 signaling pathway and viral molecules in ARPE-19 cells infected with HSV-1 were determined by qPCR and Western blotting, respectively. KEAP1-silenced and overexpressing ARPE-19 cell lines were constructed, and Western blotting was employed to assess the effects of KEAP1 silencing and overexpression on the nuclear factor erythroid 2-related factor 2 (NRF2) signaling pathway. The KEAP1-silenced and overexpressing cell lines were subsequently infected with HSV-1. Changes in viral mRNA expression were detected via qPCR, while immunofluorescence and Western blotting were used to evaluate alterations in viral protein expression. Additionally, a plaque formation assay was conducted to measure variations in viral titer. Western blotting was performed on KEAP1-silenced cell lines infected with HSV-1 to assess the expression levels of NRF2 signaling pathway and viral proteins at different time points. Results Silencing of KEAP1 activated the NRF2 signaling pathway and promoted HSV-1 replication, whereas KEAP1 overexpression downregulated the NRF2 signaling pathway and inhibited HSV-1 replication. These findings contradict previous studies suggesting that upregulation and activation of the NRF2 signaling pathway can suppress HSV-1 replication. Further investigation revealed that KEAP1 silencing-induced NRF2 upregulation was significantly inhibited following HSV-1 infection. Conclusion KEAP1 plays a crucial role in the host cell resistance to HSV-1 infection, and its interaction with NRF2 exerts complex biological functions in antiviral immune responses.

, correspAuthors=Lizu XIAO, Rongzhen LI, authorNote=null, correspAuthorsNote=, 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, authorCompany=null, fund=null, authors=, authorsList=Songbin WU, Lingfeng YE, Pengtao HU, Donglin XIONG, Lizu XIAO, Rongzhen LI), CN=ArticleExt(id=1238813313329582467, articleId=1238813310288711956, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=上调Kelch样ECH相关蛋白1 (KEAP1)抑制单纯疱疹病毒1型复制, columnId=1192149544164012138, journalTitle=微生物学报, columnName=研究报告, runingTitle=null, highlight=null, articleAbstract=

目的 探讨Kelch样ECH相关蛋白1 (Kelch-like ECH-associated protein 1, KEAP1)对单纯疱疹病毒1型(herpes simplex virus type 1, HSV-1)复制的影响，为抗单纯疱疹病毒研究提供理论依据。 方法 通过qPCR和Western blotting试验检测人视网膜色素上皮细胞(ARPE-19细胞)感染HSV-1后KEAP1-NRF2信号通路及病毒分子的mRNA和蛋白表达情况。构建KEAP1沉默和过表达的ARPE-19细胞株，采用Western blotting检测KEAP1沉默和过表达对核因子红细胞2相关因子2 (nuclear factor erythroid 2-related factor 2, NRF2)信号通路的影响。用KEAP1沉默和过表达细胞株感染HSV-1，采用qPCR检测病毒mRNA表达变化，采用免疫荧光和Western blotting检测病毒蛋白表达变化，采用空斑形成试验检测病毒滴度变化。用KEAP1沉默细胞株感染HSV-1，采用Western blotting检测不同时间点NRF2信号通路和病毒蛋白的表达情况。 结果 KEAP1沉默可激活NRF2信号通路，促进HSV-1的复制；KEAP1过表达可下调NRF2信号通路，抑制HSV-1的复制。这一结果与先前研究中关于NRF2信号通路上调及活化可抑制HSV-1复制的结论相矛盾。进一步研究发现，KEAP1沉默诱导的NRF2上调在HSV-1感染后受到显著抑制。 结论 KEAP1在宿主细胞抗HSV-1感染过程中发挥重要作用，其与NRF2的相互作用在抗病毒免疫应答中具有复杂的生物学功能。

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Herpes simplex virus 1-induced ferroptosis contributes to viral encephalitis[J]. mBio, 2023, 14(1): e0237022., articleTitle=Herpes simplex virus 1-induced ferroptosis contributes to viral encephalitis, refAbstract=null)], funds=[Fund(id=1238891101055144436, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=NS2023041, language=EN, fundingSource=Science and Technology Major Project of Shenzhen Nanshan District Health System(NS2023041), fundOrder=null, country=null), Fund(id=1238891101168390655, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=NS2023041, language=CN, fundingSource=深圳市南山区技术研发和创意设计项目(NS2023041), fundOrder=null, country=null), Fund(id=1238891101290025480, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=NS2024033, language=EN, fundingSource=Science and Technology Major Project of Shenzhen Nanshan District Health System(NS2024033), fundOrder=null, country=null), Fund(id=1238891101428437524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=NS2024033, language=CN, fundingSource=深圳市南山区技术研发和创意设计项目(NS2024033), fundOrder=null, country=null), Fund(id=1238891101516517917, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=NSZD2024029, language=EN, fundingSource=Shenzhen Nanshan District Healthcare System Science and Technology Key Project(NSZD2024029), fundOrder=null, country=null), Fund(id=1238891101629764134, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=NSZD2024029, language=CN, fundingSource=深圳市南山区卫生健康系统科技重大项目(NSZD2024029), fundOrder=null, country=null), Fund(id=1238891101763981877, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=null, language=EN, fundingSource=Municipal Financial Subsidy of Shenzhen Medical Key Discipline Construction, fundOrder=null, country=null), Fund(id=1238891101906588221, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, awardId=null, language=CN, fundingSource=深圳市医学重点学科建设项目(市级财政补贴), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1238891094801436824, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, xref=null, ext=[AuthorCompanyExt(id=1238891094809825433, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, companyId=1238891094801436824, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Pain Medicine and Shenzhen Municipal Key Laboratory for Pain Medicine, Shenzhen Nanshan People’s Hospital, Shenzhen, Guangdong, China), AuthorCompanyExt(id=1238891094818214042, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, companyId=1238891094801436824, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=深圳市南山区人民医院疼痛科，深圳市疼痛学重点实验室，广东 深圳)])], figs=[ArticleFig(id=1238891099318702440, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=EN, label=Figure 1, caption=HSV-1 infection inhibits KEAP1 expression.A: mRNA levels of KEAP1, NRF2, HO-1, ICP0, US4 and LAT after HSV-1 infection; B: Protein levels of KEAP1, NRF2, HO-1, ICP0 and gB after HSV-1 infection. **: P<0.01, ***: P<0.001, ****: P<0.000 1 vs. mock group (not infected with HSV-1), ns indicates no significant difference., figureFileSmall=ay+eT4vVSWJXOB9+DPPH1g==, figureFileBig=ip0DqR6BcnDj06OjBwRdlA==, tableContent=null), ArticleFig(id=1238891099461308788, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=CN, label=图1, caption=HSV-1感染抑制KEAP1表达。A：HSV-1感染后，KEAP1、NRF2、HO-1、ICP0、US4和LAT的mRNA水平；B：HSV-1感染后，KEAP1、NRF2、HO-1、ICP0和gB的蛋白水平。与mock组(无HSV-1感染)相比，**：P<0.01，***：P<0.001，****：P<0.000 1，ns表示无显著差异。, figureFileSmall=ay+eT4vVSWJXOB9+DPPH1g==, figureFileBig=ip0DqR6BcnDj06OjBwRdlA==, tableContent=null), ArticleFig(id=1238891099591332230, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=EN, label=Figure 2, caption=The effect of KEAP1 silencing on HSV-1 replication. A: ARPE-19 cells were transfected with KEAP1 shRNA or empty plasmid, and the expression of KEAP1 gene was detected by qPCR; B: After silencing KEAP1 using shRNAs that specifically targeted KEAP1, Western blotting was used to detect the protein level of KEAP1; C: KEAP1 silenced and control cells were infected with HSV-1 for a period of 24 h, and qPCR experiments were performed to detect the expression of virus genes ICP0, US4 and LAT; D: ICP0 and gB localization was determined by immunofluorescence assays using a fluorescence microscope to preliminarily determine the replication level of the virus; E: The expression of virus proteins ICP0 and gB were identified by Western blotting; F: Detection of viral titer in the supernatants of KEAP1 silenced cell group and control cell group by plaque assay. **: P<0.01, ***: P<0.001, ****: P<0.000 1 vs. shcontrol group., figureFileSmall=wie9bY8B7g9nDFE/qaYLYw==, figureFileBig=sKlE93FMdGhfGYVFW7LSRQ==, tableContent=null), ArticleFig(id=1238891099700384144, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=CN, label=图2, caption=KEAP1沉默对HSV-1复制的影响, figureFileSmall=wie9bY8B7g9nDFE/qaYLYw==, figureFileBig=sKlE93FMdGhfGYVFW7LSRQ==, tableContent=null), ArticleFig(id=1238891099826213271, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=EN, label=Figure 3, caption=The effect of KEAP1 overexpression on HSV-1 replication. A: ARPE-19 cells were transfected with KEAP1 plasmid or empty vector plasmid, and the expression of the KEAP1 gene was detected by qPCR; B: Western blotting was used to detect the protein level of KEAP1 after overexpression of KEAP1; C: KEAP1 overexpressing and control cells were infected with HSV-1 for a period of 24 h, and qPCR experiments were performed to detect the expression of virus genes ICP0, US4 and LAT; D: ICP0 and gB localization was determined by immunofluorescence assays using a fluorescence microscope to preliminarily determine the replication level of the virus; E: The expression of virus proteins ICP0 and gB were identified by Western blotting; F: Detection of viral titer in the supernatants of KEAP1 overexpressing cell group and control cell group by plaque assay. *: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.000 1 vs. vector group., figureFileSmall=HnJq2P/tLUjGTaGasAMw6Q==, figureFileBig=3PRe5NXKi+ybXJsfnRLHKg==, tableContent=null), ArticleFig(id=1238891099943653797, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=CN, label=图3, caption=KEAP1过表达对HSV-1复制的影响, figureFileSmall=HnJq2P/tLUjGTaGasAMw6Q==, figureFileBig=3PRe5NXKi+ybXJsfnRLHKg==, tableContent=null), ArticleFig(id=1238891100019151276, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=EN, label=Figure 4, caption=The effect of KEAP1 silencing or overexpression on the expression of NRF2 and HO-1. A: Protein levels of KEAP1, NRF2 and HO-1 after KEAP1 silencing; B: Protein levels of KEAP1, NRF2 and HO-1 after KEAP1 overexpression. *: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.000 1 vs. shcontrol or vector group., figureFileSmall=ej4oJDiBwbyO7+5JIhqWaQ==, figureFileBig=YOOdU8TWYnyZD8+n3WmbPQ==, tableContent=null), ArticleFig(id=1238891100174340533, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=CN, label=图4, caption=KEAP1沉默或过表达对NRF2和HO-1表达的影响。A：KEAP1沉默后，KEAP1、NRF2和HO-1的蛋白水平；B：过表达KEAP1后，KEAP1、NRF2和HO-1的蛋白水平。*：P<0.05，**：P<0.01，***：P<0.001，****：P<0.000 1 vs.对照组。, figureFileSmall=ej4oJDiBwbyO7+5JIhqWaQ==, figureFileBig=YOOdU8TWYnyZD8+n3WmbPQ==, tableContent=null), ArticleFig(id=1238891100442776002, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=EN, label=Figure 5, caption=HSV-1 infection suppresses NRF2 expression in shKEAP1 cell lines. Protein levels of NRF2, HO-1, ICP0, and gB in shKEAP1 cell lines following HSV-1 infection. **: P<0.01, ***: P<0.001, ****: P<0.000 1 vs. control group (not infected with HSV-1), ns indicates no significant difference., figureFileSmall=/pORkW8NBqDQTs9B021flA==, figureFileBig=RaHhTMMl39eGvyZvCJX0Mw==, tableContent=null), ArticleFig(id=1238891100585382346, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=CN, label=图5, caption=HSV-1感染抑制shKEAP1细胞株中NRF2表达。shKEAP1细胞株感染HSV-1后，NRF2、HO-1、ICP0和gB的蛋白水平。**：P<0.01，***：P<0.001，****：P<0.000 1 vs.对照组(无HSV-1感染)，ns表示无显著差异。, figureFileSmall=/pORkW8NBqDQTs9B021flA==, figureFileBig=RaHhTMMl39eGvyZvCJX0Mw==, tableContent=null), ArticleFig(id=1238891100707017172, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=EN, label=Table 1, caption=

Sequence of the primer for qPCR

, figureFileSmall=null, figureFileBig=null, tableContent=

Gene	Forward primers (5′→3′)	Reverse primers (5′→3′)
KEAP1	CTGGAGGATCATACCAAGCAGG	GGATACCCTCAATGGACACCAC
NRF2	TTCCCGGTCACATCGAGAG	TCCTGTTGCATACCGTCTAAATC
HO-1	AAGACTGCGTTCCTGCTCAAC	AAAGCCCTACAGCAACTGTCG
ICP0	GGTCGCCCTGTCGCCTTA	GGTCGCCATGTTTCCCGT
US4	TGGACACCCTCTTCG	CAGGCACACGTAACG
LAT	CGCCTTTCCTGTTCTCGCT	ACGCGGCGTCTTTGTTGA
GAPDH	TGGCCTTCCGTGTTCCTAC	GAGTTGCTGTTGAAGTCGCA

), ArticleFig(id=1238891100816069089, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813310288711956, language=CN, label=表1, caption=

qPCR引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=

Gene	Forward primers (5′→3′)	Reverse primers (5′→3′)
KEAP1	CTGGAGGATCATACCAAGCAGG	GGATACCCTCAATGGACACCAC
NRF2	TTCCCGGTCACATCGAGAG	TCCTGTTGCATACCGTCTAAATC
HO-1	AAGACTGCGTTCCTGCTCAAC	AAAGCCCTACAGCAACTGTCG
ICP0	GGTCGCCCTGTCGCCTTA	GGTCGCCATGTTTCCCGT
US4	TGGACACCCTCTTCG	CAGGCACACGTAACG
LAT	CGCCTTTCCTGTTCTCGCT	ACGCGGCGTCTTTGTTGA
GAPDH	TGGCCTTCCGTGTTCCTAC	GAGTTGCTGTTGAAGTCGCA

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