Article(id=1217471086472642705, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1217471079325549522, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250547, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1752681600000, receivedDateStr=2025-07-17, revisedDate=null, revisedDateStr=null, acceptedDate=1755878400000, acceptedDateStr=2025-08-23, onlineDate=1768197326533, onlineDateStr=2026-01-12, pubDate=1767456000000, pubDateStr=2026-01-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768197326533, onlineIssueDateStr=2026-01-12, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768197326533, creator=13701087609, updateTime=1768197326533, updator=13701087609, issue=Issue{id=1217471079325549522, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='1', pageStart='1', pageEnd='475', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768197324830, creator=13701087609, updateTime=1768198886678, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1217477630291530315, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1217471079325549522, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1217477630291530316, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1217471079325549522, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=92, endPage=103, ext={EN=ArticleExt(id=1217471086724300965, articleId=1217471086472642705, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Research progress in the antiviral effects and mechanisms of chloroquine, columnId=1192149543727808575, journalTitle=Acta Microbiologica Sinica, columnName=Review, runingTitle=null, highlight=null, articleAbstract=

Chloroquine, a low-cost antimalarial agent, has garnered significant interest due to its extensive research foundation and potential anti-tumor and antiviral properties. Chloroquine exhibits broad-spectrum inhibitory effects against diverse human and animal pathogenic viruses in vitro. Its antiviral efficacy has been demonstrated against Zika virus and feline coronavirus in vivo. The primary action mechanisms of chloroquine include inhibition of viral binding to host cells and subsequent internalization, modulation of viral nucleic acid recognition pathways, blockade of autophagosome maturation, and regulation of cytokine secretion in the immune response. This review systematically summarizes the antiviral effects and mechanisms of chloroquine, providing a theoretical foundation for the future development of chloroquine and its derivatives as antivirals.

, correspAuthors=Chao YE, authorNote=null, correspAuthorsNote=
*E-mail:
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氯喹是一种价格低廉的传统抗疟药物,因其广泛的研究基础以及潜在的抗肿瘤、抗病毒特性而备受关注。氯喹在体外对多种人类和动物致病病毒具有广谱抑制作用,在体内对寨卡病毒以及猫冠状病毒也具有抗病毒效果。氯喹的作用机制主要包括:抑制病毒与宿主细胞的结合及内化过程,调控病毒核酸识别通路,阻断自噬体成熟,以及调节细胞因子分泌等免疫应答。本文系统综述了氯喹的抗病毒作用及其分子机制,为后续将氯喹及其衍生物开发为抗病毒药物提供了理论参考。

, correspAuthors=叶超, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=yba61azrTH3LoQXVesNMOQ==, magXml=yKgWl0RN/CrGK/7xEWUylw==, pdfUrl=null, pdf=MJivNNIUrZ0oqOozNNZI9g==, pdfFileSize=757384, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=cqDhEfrJlTXgreFGk8qNwQ==, mapNumber=null, authorCompany=null, fund=null, authors=

作者贡献声明

李浩林:提出概念,撰写文章,撰写、审阅;代宇:提出概念,审阅;方仁东:审阅,获取基金;叶超:获取基金,撰写、审阅。

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氯喹抗病毒作用及机制研究进展
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李浩林 , 代宇 , 方仁东 , 叶超 *
微生物学报 | 综述 2026,66(1): 92-103
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微生物学报 | 综述 2026, 66(1): 92-103
氯喹抗病毒作用及机制研究进展
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李浩林, 代宇, 方仁东, 叶超*
作者信息
  • 西南大学 动物医学院,动物健康与动物性食品安全国际合作联合实验室,重庆
Research progress in the antiviral effects and mechanisms of chloroquine
Haolin LI, Yu DAI, Rendong FANG, Chao YE*
Affiliations
  • Joint International Research Laboratory of Animal Health and Animal Food Safety, College of Veterinary Medicine, Southwest University, Chongqing, China
出版时间: 2026-01-04 doi: 10.13343/j.cnki.wsxb.20250547
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氯喹是一种价格低廉的传统抗疟药物,因其广泛的研究基础以及潜在的抗肿瘤、抗病毒特性而备受关注。氯喹在体外对多种人类和动物致病病毒具有广谱抑制作用,在体内对寨卡病毒以及猫冠状病毒也具有抗病毒效果。氯喹的作用机制主要包括:抑制病毒与宿主细胞的结合及内化过程,调控病毒核酸识别通路,阻断自噬体成熟,以及调节细胞因子分泌等免疫应答。本文系统综述了氯喹的抗病毒作用及其分子机制,为后续将氯喹及其衍生物开发为抗病毒药物提供了理论参考。

氯喹  /  病毒  /  机制

Chloroquine, a low-cost antimalarial agent, has garnered significant interest due to its extensive research foundation and potential anti-tumor and antiviral properties. Chloroquine exhibits broad-spectrum inhibitory effects against diverse human and animal pathogenic viruses in vitro. Its antiviral efficacy has been demonstrated against Zika virus and feline coronavirus in vivo. The primary action mechanisms of chloroquine include inhibition of viral binding to host cells and subsequent internalization, modulation of viral nucleic acid recognition pathways, blockade of autophagosome maturation, and regulation of cytokine secretion in the immune response. This review systematically summarizes the antiviral effects and mechanisms of chloroquine, providing a theoretical foundation for the future development of chloroquine and its derivatives as antivirals.

chloroquine  /  viruses  /  mechanisms
李浩林, 代宇, 方仁东, 叶超. 氯喹抗病毒作用及机制研究进展. 微生物学报, 2026 , 66 (1) : 92 -103 . DOI: 10.13343/j.cnki.wsxb.20250547
Haolin LI, Yu DAI, Rendong FANG, Chao YE. Research progress in the antiviral effects and mechanisms of chloroquine[J]. Acta Microbiologica Sinica, 2026 , 66 (1) : 92 -103 . DOI: 10.13343/j.cnki.wsxb.20250547
传染病是人类发病与死亡的主要原因之一,也是阻碍全世界人民发展与福祉提升的巨大障碍。在困扰全球的传染病疫情中病毒病占一半以上[1]。开发具有广谱抗病毒活性的药物对于病毒病疫情的防控具有重要意义。氯喹(chloroquine, CQ)作为疟疾的传统治疗药物已在临床应用多年。由于其对自噬的抑制作用以及其免疫调节等特性,氯喹显示出治疗癌症和病毒性传染病的潜力[2],并被报道为一种潜在的广谱抗病毒药物。它可在体外抑制多种病毒的复制,如埃博拉病毒(ebola virus, EBOV)、基孔肯雅热病毒(chikungunya virus, CHIKV)、寨卡病毒(Zika virus, ZIKV)等,并且氯喹在小鼠体内对寨卡病毒、在猫体内对猫传染性腹膜炎病毒(feline infectious peritonitis virus, FIPV)均显示出抗感染活性。因此氯喹具有良好的抗病毒应用前景。
氯喹类药物属于4-氨基喹啉衍生物,氯喹的硫酸盐、磷酸盐均已被制成商品化的抗疟药[3],主要通过在疟原虫食物泡中积累,升高pH值来发挥抗疟作用[4]。此外,氯喹还可用于治疗系统性红斑狼疮(systemic lupus erythematosus, SLE)、类风湿性关节炎(rheumatoid arthritis, RA)等自身免疫性疾病[5]。通过对氯喹进行结构修饰研究,在其结构中引入羟基合成了羟氯喹(hydroxychloroquine, HCQ),研究发现羟氯喹的毒性较氯喹更低,作为氯喹的替代品也得到了广泛应用[6]。氯喹和羟氯喹均可被胃肠道快速吸收,口服后2-3 h血药浓度即可达到峰值,且能渗透到大多数组织中,生物利用度可达70%-80%[7]。氯喹是一种二元弱碱,能以带电(质子化)和不带电(去质子化) 2种形式存在。去质子化的氯喹可自由、快速地扩散穿过细胞膜和细胞器膜[8]。一旦质子化,氯喹便无法再自由扩散,会滞留在pH值较低的内体、高尔基体和溶酶体等细胞器中,升高这些细胞器的pH值,影响细胞器中酶的活性,进而影响其生物学功能,最终产生抗病毒效应。
埃博拉病毒(Ebola virus, EBOV)是一种具有包膜的单股负链RNA丝状病毒,属于丝状病毒科(Filoviridae),主要通过体液接触传播,具有高死亡率[9]。Dowall等[10]发现氯喹可减少人肺原代成纤维细胞(medical research council cell strain-5, MRC-5)中EBOV的复制,但无法预防豚鼠感染EBOV。Falzarano等[11]证实,以对氯喹的最大耐受剂量(90 mg/kg)治疗EBOV感染的小鼠并未改变其存活率,在仓鼠模型中由于氯喹毒性较大,较高剂量(90 mg/kg)的氯喹会直接导致仓鼠死亡;当以较低剂量(50 mg/kg)的氯喹与多西环素和阿奇霉素合用时仓鼠存活率也未发生改变。尽管氯喹在体外细胞实验中显示出抑制EBOV复制的效果,但在多种动物模型(豚鼠、小鼠、仓鼠)的体内试验中氯喹均未能表现出预防感染或提高存活率的治疗效果。
基孔肯雅热病毒(chikungunya virus, CHIKV)是一种有包膜的单股正链RNA病毒,属于甲病毒科(Alphavirus),起源于非洲,自2005年以来,该病毒在世界范围内多次暴发[12]。CHIKV在感染的晚期阶段可于巨噬细胞(如滑膜巨噬细胞)中持续存在,导致组织损伤、凋亡、纤维化和极化炎症反应[13]。Roques等[14]证实无论使用何种浓度(1-50 μmol/L)的氯喹均可抑制CHIKV在巨噬细胞中的复制,但经氯喹处理的几只猕猴在感染CHIKV后症状更加严重。氯喹在体外细胞实验中能够抑制CHIKV在易感细胞中的复制,但在以猕猴进行的体内试验中氯喹未表现出治疗效果,反而加重了症状。氯喹在其他实验中显示出抑制I型干扰素(type I interferon, IFN-I)的作用[15],而IFN-I对CHIKV复制有抑制作用[16],这可能是导致氯喹在体内实验中无法产生抗CHIKV效果的原因。
寨卡病毒(Zika virus, ZIKV)是一种有包膜的单股正链RNA病毒,属于黄病毒属(Flavivirus),具有嗜神经毒性,每年导致数百万例感染[17]。感染ZIKV的母亲所生儿童的出生缺陷率比正常人高出约20倍[18]。Delvecchio等[19]发现氯喹以剂量依赖性方式抑制了Vero细胞中ZIKV的复制,且主要对病毒复制周期的早期有抑制作用,使用氯喹处理神经祖细胞形成的神经球时,ZIKV的复制也被抑制。缺乏I型(α/β)和II型(γ) IFN受体的AG129小鼠感染ZIKV后症状最严重,是研究ZIKV体内感染的理想小鼠模型[20],氯喹延长了被ZIKV感染的AG129小鼠的平均寿命且减轻了小鼠的症状[17]。Swiss Jim Lambert (SJL)小鼠模型常用于研究高剂量ZIKV感染引发的胎儿传播[21],可用于测试可抑制病毒垂直传播的药物。氯喹处理ZIKV感染的SJL小鼠可使母体血液和胎儿大脑中的ZIKV滴度降低约95%[17]。氯喹不仅在体外显示出抑制ZIKV的能力,而且在AG129小鼠模型中也能有效抑制ZIKV复制并减轻感染症状,尤其能显著降低母婴传播模型中胎儿的病毒载量,显示出潜在的治疗价值。
登革热病毒(dengue virus, DENV)是一种有包膜的单链RNA病毒,属于黄病毒属(Flavivirus),是全球分布最广泛的虫媒病毒,目前近半数世界人口面临感染登革热病毒的风险[22]。氯喹可抑制DENV在U937细胞和Vero细胞中的复制,但不会抑制DENV在白纹伊蚊(Aedes albopictus) C6/36细胞中的复制[23-24]。Farias等[25]观察到氯喹可缩短感染DENV的夜猴(Aotus trivirgatus)的病毒血症持续时间,但夜猴模型不会像人类感染DENV那样出现登革热(dengue fever, DF)、登革出血热(dengue hemorrhagic fever, DHF)以及登革休克综合征(dengue shock syndrome, DSS)等症状,因此该结果不足以充分证明氯喹在体内具有显著的抗病毒作用。Borges等[26]研究表明氯喹治疗3 d可改善登革热患者的症状,但不会改变疾病或发烧的持续时间。Tricou等[27]同样使用氯喹治疗3 d,治疗组患者的病毒血症清除时间、退烧时间以及炎性细胞因子等相较于对照组均无显著差异。因此氯喹在体内对登革热病毒可能仅具有轻微甚至无抗病毒活性。
严重急性呼吸综合征冠状病毒(severe acute respiratory syndrome coronavirus, SARS-CoV)是有包膜的单股正链RNA病毒[28],属于Beta冠状病毒属(Betacoronavirus),于2002年首次出现后的几个月内迅速传播到至少30个国家,最终确定病原体为SARS-CoV[29]。Vincent等[30]在SARS-CoV感染Vero细胞前用不同浓度的氯喹预处理,结果显示病毒抗原阳性细胞以剂量依赖性减少,并且10 μmol/L的氯喹可完全抑制SARS-CoV感染。在病毒感染后加入氯喹,病毒抗原阳性细胞的数量急剧下降,且感染几乎不会向邻近细胞扩散。Barnard等[31]用不同浓度的氯喹预处理小鼠,结果显示不同剂量的氯喹不具备对SARS-CoV的复制产生影响。虽然氯喹在体外模型中可有效抑制SARS-CoV复制与扩散,但在体内实验未观察到其对SARS-CoV的显著抗病毒效应。
严重急性呼吸系统综合征冠状病毒2型(severe acute respiratory syndrome coronavirus 2, SARS-CoV-2)是一种有包膜的单股正链RNA病毒,属于Beta冠状病毒属(Betacoronavirus),潜伏期短、传播迅速,较难控制,且感染后会导致患者死亡[32]。Wang等[33]使用氯喹对SARS-CoV-2感染的Vero细胞进行全过程处理、入侵前处理、入侵后处理,抑制率分别达到了80%、60%、40%,这表明氯喹主要在SARS-CoV-2的侵入阶段发挥作用。尽管氯喹在体外可有效抑制SARS-CoV-2的复制,但目前相关研究结果显示氯喹对SARS-CoV-2的无症状感染者和有症状感染者几乎无作用[34-35]
单纯疱疹病毒1型(herpes simplex virus type 1, HSV-1)是有包膜的双链DNA病毒,属于疱疹病毒科(Herpesviridae),可引发原发性和复发性水疱性病变[36]。HSV-1穿入细胞以及细胞与细胞之间的传播都需要糖蛋白D (glycoprotein D, gD),其对HSV-1的感染具有重要作用。Singh等[37]研究表明虽然氯喹(150 μmol/L)不能有效阻止HSV对细胞的吸附或侵入,并且在亚效应剂量下氯喹和干扰素单独使用不会阻断HSV-1的gD蛋白的运输,也不会对HSV-1的感染性产生影响,但当它们联合使用时gD蛋白在药物处理过的细胞中出现积累,并且HSV-1的感染性也降低。Maheshwari等[38]发现干扰素会提高高尔基体反面囊膜的pH值,阻断蛋白从反式高尔基体到质膜的运输。氯喹可能通过增强这一过程增强了干扰素对gD蛋白运输的阻断作用。
猪流行性腹泻病毒(porcine epidemic diarrhea virus, PEDV)是有包膜的单股正链RNA病毒,属于α冠状病毒属(Alphacoronavirus),可引起新生仔猪的急性腹泻、呕吐、脱水和高死亡率[39]。Wang等[40]证实PEDV感染显著增加了自噬体和自噬溶酶体的数量,氯喹处理减少了PEDV感染的Vero细胞中自噬溶酶体的数量,并降低了Vero细胞裂解液中的猪流行性腹泻病毒S蛋白和N蛋白水平以及病毒滴度。
牛病毒性腹泻病毒(bovine viral diarrhea virus, BVDV)是有包膜的单股正链RNA病毒,属于黄病毒科(Flaviviridae),可导致牛出现腹泻、呼吸道症状以及生殖功能障碍[41]。研究显示BVDV可以诱导自噬[42]。Wang等[43]在使用氯喹处理阻断牛肾细胞(madin-darby bovine kidney, MDBK)的自噬时,BVDV E2蛋白的表达减少,病毒复制受到抑制。
猫杯状病毒(feline calici virus, FCV)是无包膜的单股正链RNA病毒,属于杯状病毒科(Caliciviridae),具有高度传染性,高毒力FCV可诱发严重的全身性疾病,并伴有高死亡率[44]。Kreutz等[45]证明在FCV感染猫肾细胞的初期阶段加入氯喹能够抑制感染性病毒的产生,但如果在感染后2 h加入则不能产生明显抑制作用,并且氯喹的作用是可逆的,当从培养基中移除氯喹后病毒复制可以继续进行。Stuart等[46]也表明氯喹并不会影响FCV与细胞的结合,而是在病毒结合后、病毒RNA释放之前的早期阶段起作用。因此氯喹可能主要通过抑制FCV的进入发挥抗病毒作用。
猫冠状病毒(feline coronavirus, FCoV)是有包膜的单股正链RNA病毒,属于α冠状病毒属(Alphacoronavirus),FCoV有2种血清型,即猫肠道冠状病毒(feline enteric coronavirus, FECV) (无毒FCoV)和猫传染性腹膜炎病毒(feline infectious peritonitis virus, FIPV) (毒性FCoV),FIPV是一种致死性强的病毒,并且目前对FIPV感染尚无有效疫苗和特异性治疗药物[47]。Takano等[48]证实氯喹显著抑制单核细胞中FIPV的增殖以及炎性细胞因子mRNA的水平,然而用氯喹处理已经感染FIPV的细胞并不会产生抗病毒效果;FIPV感染猫经氯喹治疗后的临床评分优于未治疗组,这表明氯喹对感染FIPV的猫具有治疗效果。
唾液酸(sialic acid, SA)是一类含有9碳骨架的酸性糖类物质的总称,广泛存在于真核生物细胞的细胞表面糖蛋白、糖脂和游离寡糖的糖链非还原末端[49]。SA参与多种病毒感染和传播周期的多个阶段。许多病毒利用SA末端聚糖作为主要受体或辅助受体介导与宿主细胞结合和内化[50]。Helander等[51]证明SA对于呼肠孤病毒(reovirus, ReoV) T1L血清型黏附兔M细胞具有重要作用;Desforges等[52]表明SA可以与人冠状病毒OC43 (human coronavirus OC43, HCoV-OC43)的S蛋白发生相互作用促进HCoV-OC43的吸附;Liu等[53]证明SA作为人类肠道病毒D68 (human enterovirus D68, EV-D68)的功能性细胞受体,诱导病毒构象变化以启动病毒脱壳和进入细胞。氯喹被证明可以抑制参与SA生物合成的醌还原酶2 (quinone oxidoreductase 2, QR2)[54]。因此氯喹可通过抑制QR2干扰唾液酸的合成抑制病毒进入前步骤。
SARS-CoV可通过结合易感细胞膜上的血管紧张素转换酶2 (angiotensin converting enzyme 2, ACE2)受体感染细胞[55]。Vincent等[30]证明在氯喹的有效抗SARS-CoV浓度下细胞表面的ACE2表达量无显著变化,但其末端糖基化受损。氯喹预处理可能导致糖基化不足的ACE2在细胞表面表达。Wang等[56]证实氯喹和羟氯喹会与ACE2结合,这表明氯喹可能通过竞争性抑制来阻止SARS-CoV-2和ACE2的结合,从而抑制病毒感染。因此氯喹预处理可能导致细胞表面表达糖基化不足的ACE2,并通过竞争性抑制阻止病毒和ACE2的结合,降低其与病毒刺突蛋白的结合,从而抑制病毒感染易感细胞。
病毒可通过经内吞作用进入胞内的内体,进入内体后许多病毒会因pH值下降和酸性内体蛋白酶(尤其是组织蛋白酶L和组织蛋白酶B)的水解切割导致穿透能力变化,触发穿透过程进入胞质[57]。氯喹可使溶酶体pH值升高[58],抑制了需要酸化环境才能从内体到达细胞质的病毒感染过程。ZIKV的E蛋白与宿主细胞受体结合可促进病毒颗粒与内体膜融合,释放到宿主细胞细胞质中[59]。Zhang等[60]发现在用氯喹处理的Vero细胞中大多数ZIKV与早期内体共定位,极少数病毒颗粒与溶酶体共定位,证实氯喹阻止了病毒从内体释放到细胞质进行复制的过程,抑制了ZIKV的复制。SARS-CoV-2的S蛋白在蛋白酶的作用下暴露出S2融合肽区,促进病毒囊膜和内体膜发生融合[61],从而将病毒基因组释放到胞质中,而氯喹可抑制内体酸化过程,这可能会抑制溶酶体蛋白酶的活性,从而干扰病毒和宿主膜的融合[62]。氯喹还可直接抑制病毒内化发挥抗病毒作用,Diaz-Griffero等[63]证实禽白血病病毒(avian leukosis virus, ALV) B亚型是需要内吞作用才能到达内体的pH依赖性病毒,使用氯喹可将进入细胞的ALV-B减少了79.8%。因此氯喹可通过抑制病毒释放到胞质和内化过程发挥抗病毒作用。
Chiang等[64]表明氯喹可抑制T细胞和单核细胞中人类免疫缺陷病毒(human immunodeficiency virus, HIV)-1糖蛋白-gp120的翻译后修饰。Randolph等[65]也证实氯喹会抑制黄病毒(flavivirus, FLAV) prM蛋白水解加工为M蛋白。因此氯喹可通过升高酸性区室(如内质网、反式高尔基体网络)的pH值抑制依赖酸性环境的蛋白酶与糖基转移酶活性,从而干扰病毒蛋白的翻译后修饰(如糖基化、蛋白水解)及病毒颗粒组装[66]。氯喹也可抑制高尔基体的出芽过程。在HSV-1模型中氯喹会升高高尔基体囊泡的pH[67],并抑制高尔基体的出芽,导致非感染性HSV-1颗粒在反式高尔基体网络中积累[68-69],从而抑制合成的病毒蛋白的翻译后修饰。
机体的先天免疫系统可以通过模式识别受体(pattern recognition receptor, PRR)识别病毒的病原体相关分子模式(pathogen-associated molecular patterns, PAMPs)。Toll样受体(Toll-like receptors, TLR)是研究最深入的PRR之一,位于内体的TLR (主要是TLR3、TLR7、TLR9)可识别病毒的单链RNA (single-stranded RNA, ssRNA)、双链RNA (double-stranded RNA, dsRNA)和富含CpG岛的DNA,在机体的先天免疫应答中发挥着重要作用;TLR识别病毒核酸后会激活核因子κB (nuclear factor kappa B, NF-κB)等激酶,最终调节细胞因子、趋化因子和IFN的表达[70]。TLR3、TLR7和TLR9在细胞内表达于内质网、内体、多泡体和溶酶体中,激活仅发生在酸化的内体,而氯喹可以通过升高内体pH影响TLR的激活过程[71-72],氯喹还可以通过结合内体内的病毒核酸阻止内体TLR与其配体的相互作用[73],从而干扰TLR下游信号通路。
细胞质DNA感受器环鸟苷酸-腺苷酸合成酶(cyclic GMP-AMP synthase, cGAS)可以识别细胞质中的双链DNA,催化合成第二信使2′3′-环鸟苷酸-腺苷酸(2′3′-cyclic-GMP-AMP, cGAMP)[74]。cGAMP随后结合并激活内质网驻留蛋白干扰素基因刺激因子(stimulator of interferon genes, STING),激活的STING转位至高尔基体,募集并激活抑制因子激酶(inhibitor of kappa B kinase, IKK)和TANK结合激酶1 (TANK-binding kinase 1, TBK1),分别磷酸化激活NF-κB和干扰素调节因子3 (interferon regulatory factor 3, IRF3)。这些转录因子入核诱导IFN-I和细胞因子表达,介导抗病原体免疫[75-77]。氯喹会显著抑制HSV-1诱导的cGAS的降解[78],Ma等[79]发现伪狂犬病病毒(pseudorabies virus, PRV)的UL21通过自噬-溶酶体途径降解cGAS抑制cGAS-STING1介导的先天免疫,并且氯喹抑制了这个过程。Jiao等[80]证实氯喹显著减弱了SARS-CoV-2的非结构蛋白6 (nonstructural protein 6, NSP6)对STING1表达的抑制效果,并消除了NSP6在HEK293T细胞中对cGAS-STING1激活的IFN-β表达的抑制作用。因此氯喹可通过抑制病毒感染中cGAS-IFN-I信号通路关键蛋白的降解维持cGAS-IFN-I信号通路激活。
视黄酸诱导基因-I样受体(RIG-I-like receptor, RLR)在几乎所有有核细胞中表达,是识别RNA病毒的主要胞质PRR,是抵御多种病毒家族的第一道防线[81-82]。RIG-I caspase募集结构域(caspase recruitment domain, CARD)和MDA5 CARD通过CARD-CARD介导的相互作用与线粒体抗病毒信号转导蛋白(mitochondrial antiviral signaling protein, MAVS)结合,最终激活NF-κB和IFN-I信号通路[83]。Liu等[84]发现氯喹完全抑制了稳态铁调节因子(hemochromatosis protein, HFE)蛋白(抗病毒先天免疫的负调节因子)通过选择性自噬-溶酶体途径介导的MAVS降解。猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus, PRRSV)的非结构蛋白Nsp5会依赖自噬-溶酶体途径诱导RLR信号通路的MAVS、TBK1、IRF3和IRF7等多个关键蛋白降解,而氯喹抑制了Nsp5介导的RLR信号通路蛋白降解[85]。因此氯喹可通过抑制自噬-溶酶体途径介导的RLR信号通路关键蛋白的降解,增强针对RNA病毒感染的先天免疫反应。
自噬体与内体和溶酶体的囊泡融合通过酸性蛋白酶清除被自噬体吞噬的物质[86]。自噬是免疫的主要参与者,它通过调节IFN的产生和炎性小体活性来减轻炎症[87],还可通过降解病毒粒子发挥抗病毒作用[86]。自噬能够清除细胞内病原体,然而自噬又是一把“双刃剑”,在某些情况下会促进病毒的存活和复制。O’Donnell等[88]证明口蹄疫病毒(foot-and-mouth disease virus, FMDV)会触发自噬,并促进病毒进入细胞,进而增强病毒复制。人类B19细小病毒在其具备病毒复制能力之前诱导自噬,从而促进感染细胞的存活,使其能够在细胞崩溃之前完成病毒复制[89]。自噬体可作为脊髓灰质炎病毒(poliovirus, PV)复制的膜性支撑结构,也可作为小鼠肝炎病毒(mouse hepatitis virus, MHV)复制的位点[90],最终促进病毒复制。氯喹可沿pH梯度进入溶酶体并在溶酶体中积累,通过升高pH值抑制溶酶体酶的活性,从而抑制自噬溶酶体中货物的降解[91]。氯喹还会影响自噬体-溶酶体融合,Mauthe等[92]通过观察氯喹处理后的细胞中LC3与LAMP2的共定位情况证实氯喹损害了自噬体与溶酶体的融合。这最终可抑制依赖宿主自噬促进自身复制的病毒。进入细胞后的日本脑炎病毒(Japanese encephalitis virus, JEV)会被靶向运送至自噬溶酶体进行病毒脱壳,Li等[93]敲低自噬必需基因自噬相关蛋白5 (autophagy related 5, ATG5)以及使用磷脂酰肌醇-3激酶(phosphatidylinositol-3-kinase, PI3K)抑制剂3-甲基腺嘌呤(3-methyladenine, 3-MA)后LC3-II水平降低,JEV复制减少。Zhang等[94]发现在JEV感染小鼠模型中使用氯喹处理可导致脑组织中LC3与JEV共定位的神经元数量减少,并减轻了小鼠的症状,因此氯喹可通过阻断自噬抑制病毒的复制。ZIKV感染可高效诱导LC3-I向LC3-II转化,促进自噬。Zhang等[60]发现ZIKV在ATG5缺陷的小鼠胚胎成纤维细胞(mouse embryonic fibroblasts, MEFs)中的复制完全被抑制,而氯喹可逆转ZIKV诱导的p62降解,并且在野生型MEFs中对ZIKV产生的抑制效果与ATG5缺陷MEFs中对ZIKV复制的抑制效果相当。
病毒感染宿主会导致不同的免疫细胞异常激活,这些激活的免疫细胞产生大量促炎细胞因子和趋化因子,会通过正反馈进一步促进其他免疫细胞的激活[95],而这种过度激活的免疫反应会对宿主产生负面影响。在SARS-CoV-2导致的细胞因子风暴中促炎细胞因子水平升高并伴有广泛的肺部炎症和肺部损伤[96]。Tang等[95]认为抗病毒药物和适当的免疫调节疗法(控制过度激活的炎症反应)的组合可能是对抗COVID-19的重要策略。Satarker等[97]认为羟氯喹可通过预防细胞因子风暴抑制SARS-CoV-2的复制。因此氯喹可能通过调控过度激活的炎症反应在减轻病毒感染对宿主的进一步损伤方面发挥重要作用。氯喹能以剂量依赖性方式诱导细胞周期蛋白依赖性激酶抑制剂p21的转录,抑制CD4+ T细胞增殖,还会通过影响c-Jun氨基末端激酶(c-Jun N-terminal kinase, JNK)/转录因子活化蛋白-1 (activator protein-1, AP-1)信号传导影响T细胞亚群细胞因子的分泌,显著减少Th1细胞分泌IFN-γ和Th17细胞分泌白细胞介素-17 (interleukin-17, IL-17)[98-99]。氯喹还以剂量依赖性方式抑制TLR9诱导的B细胞增殖、浆母细胞形成以及IgG的分泌,并对分泌IL-10的调节性B细胞(regulatory B cell, Breg)的抑制作用较颗粒酶B阳性B细胞(granzyme B-secreting B cells, GrB-secreting B cells)更为显著,其在临床浓度下可强烈抑制所有B细胞亚群产生诸如IL-6、IL-1β和TNF-α等炎症细胞因子[100-101]。由此可见,氯喹通过其免疫调节作用可减少病毒感染过程中的免疫细胞过度激活,从而抑制细胞因子风暴并减轻宿主损伤。
寻找价格低廉的抗病毒药物对于兽医领域病毒性传染病的防控至关重要。本课题组Dai等[102]研究发现,抗疟药氯喹能显著抑制PRV在猪肾细胞(porcine kidney-15, PK-15)中的感染,并通过阻断病毒吸附和内化发挥抗病毒作用;该研究通过阐明氯喹抗伪狂犬病病毒的作用与机制为伪狂犬病的药物防治奠定了理论基础。因此本文对氯喹的抗病毒作用和抗病毒机制进行了梳理,希望为深入研究和开发氯喹的抗病毒作用提供理论参考。
有研究显示氯喹与大环内酯类药物联合使用并不能对感染SARS-CoV-2的患者起到治疗作用,但Singh等[37]研究表明氯喹会增强干扰素对HSV-1的抑制作用,因此氯喹也可能通过与其他药物联合用药取得良好的抗病毒作用。大量实验表明氯喹在体外显示出抗病毒作用,并且主要是在病毒感染的吸附和侵入阶段发挥作用。将氯喹开发为靶向病毒感染早期的抗病毒药物,并通过联用其他抗病毒药物可能获得更好的疗效。氯喹在发挥抗病毒作用的同时,还会对机体产生免疫抑制作用并抑制炎性细胞因子的释放,而氯喹对于机体的免疫抑制作用可能正是其体内抗病毒作用不理想的原因。寻找与氯喹联用的药物以平衡氯喹的抗病毒与免疫抑制作用可提升其体内抗病毒效果。对氯喹分子本身进行结构优化,或许也能实现氯喹的体内抗病毒作用。其衍生物羟氯喹就是一个成功先例,它作为抗疟药具有更低毒性和更高安全性。通过进一步的结构修饰降低氯喹对肝、肾及心脏等器官的毒性,有望在提高临床使用剂量的同时实现更佳的体内抗病毒效果。
在成年人中氯喹的剂量超过20 mg/kg即表现出毒性,超过40 mg/kg可致命[103]。相比之下,小鼠可耐受90 mg/kg的氯喹,仓鼠能耐受50 mg/kg的氯喹[11]。这表明不同物种对氯喹的耐受性存在显著差异。目前针对大型经济动物(如家畜)的氯喹药代动力学研究仍较为缺乏。现有动物实验中所观察到的耐受剂量普遍高于人类耐受剂量,因此大型经济动物可能对氯喹具有更高的耐受剂量,这可能使氯喹的体内抗病毒作用更易实现,这仍需通过进一步的药代动力学与药效学实验加以验证。此外,药代动力学研究显示,氯喹主要分布在脑脉络丛区域、心脏浦肯野细胞、支气管上皮细胞、II型肺泡细胞以及肺泡壁表面[104],提示氯喹可能对主要感染脑、支气管以及肺的病毒更易实现体内抗病毒作用。
  • 国家自然科学基金(32372982)
  • 国家生猪技术创新中心项目(NCTIP-XD/C17)
  • 重庆现代农业产业技术体系(CQMAITS202512)
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doi: 10.13343/j.cnki.wsxb.20250547
  • 接收时间:2025-07-17
  • 首发时间:2026-01-12
  • 出版时间:2026-01-04
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  • 收稿日期:2025-07-17
  • 录用日期:2025-08-23
基金
National Natural Science Foundation of China(32372982)
国家自然科学基金(32372982)
National Center of Technology Innovation for Pigs(NCTIP-XD/C17)
国家生猪技术创新中心项目(NCTIP-XD/C17)
Chongqing Modern Agricultural Industry Technology System(CQMAITS202512)
重庆现代农业产业技术体系(CQMAITS202512)
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    西南大学 动物医学院,动物健康与动物性食品安全国际合作联合实验室,重庆

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
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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