Article(id=1212692427429827341, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212692423956939344, articleNumber=1001-2494(2024)17-1581-09, orderNo=null, doi=10.11669/cpj.2024.17.005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1715356800000, receivedDateStr=2024-05-11, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1767058005435, onlineDateStr=2025-12-30, pubDate=1725724800000, pubDateStr=2024-09-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1767058005435, onlineIssueDateStr=2025-12-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1767058005435, creator=13701087609, updateTime=1767058005435, updator=13701087609, issue=Issue{id=1212692423956939344, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='17', pageStart='1553', pageEnd='1664', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1767058004596, creator=13701087609, updateTime=1767058886858, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1212696124457140722, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212692423956939344, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1212696124457140723, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212692423956939344, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1581, endPage=1589, ext={EN=ArticleExt(id=1212692427807314704, articleId=1212692427429827341, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Coptisine Derivative Q3 Ameliorates DSS-induced Ulcerative Colitis in Mice by Modulating TLR4/NF-κB Signaling Pathway and Intestinal Microbiota, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To explore the mechanism of action of coptisine derivative Q3 against dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice from the perspective of changes in the inflammatory signaling pathway TLR4/NF-κB and compositions of the intestinal microbiota. METHODS Fifty mice were randomly divided into normal control group, model group, sulfasalazine (SASP) group, Q3-low dose group, Q3-high dose group, with 10 in each group. Mice in the groups except the control group were orally administered with 2.5% DSS solution to induce UC model. The SASP was given 700 mg·kg^-1·d^-1 of sulfasalazine by intragastric administration, and the Q3 low and high dose groups were given 50 and 100 mg·kg^-1·d^-1 of Q3, respectively. The other groups were given an equal amount of distilled water. After 6 days of administration, the mouse colon tissues were taken for DAI score and length measurement. HE staining was used to detect the degree of pathological damage in each group. 16S rRNA high-throughput sequencing was used to detect changes in intestinal flora in the intestinal contents of mice. Immunohistochemistry and Western blot were used to detect the expression of TLR4 and p-p65 in colon tissue. The protein expressions of TLR4, p-p65 and p-IκBα and the nuclear translocation of NF-κB p65 in IEC6 cells and RAW264.7 cells were detected by Western blot and immunofluorescence. RESULTS In the DSS-induced mouse ulcerative colitis model, compared with the model group, in vivo Q3 could significantly improve the weight loss, colon length, and increase in DAI scores of UC mice. HE staining results showed that Q3 significantly improved the intestinal pathological damages such as tract epithelial damage, crypt structure disorder and goblet cell reduction; immunohistochemistry and Western blot results showed that Q3 could significantly reduce the expression of TLR4 and p-p65 in the colon tissue of mice in the model group. The results of 16S rRNA showed that Q3 could increase the biodiversity of intestinal microbiota and regulate the composition of intestinal microbiota after DSS administration. It was also shown that in vitro Q3 could inhibit the nuclear translocation of NF-κB p65 in both IEC6 and RAW264.7 cells by immunofluorescence. CONCLUSION Q3 can improve intestinal inflammation by inhibiting the TLR4/NF-κB pathway and regulating the composition of intestinal flora, exerting anti-UC effects, which is expected to become a candidate compound for the treatment of UC.

, correspAuthors=Lianqiu WU, 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, authorCompany=null, fund=null, authors=null, authorsList=Xiaoqian CHI, Changwei CHAI, Haijing ZHANG, Lianqiu WU), CN=ArticleExt(id=1212692430021907283, articleId=1212692427429827341, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=黄连碱衍生物Q3通过调节TLR4/NF-κB信号通路和肠道微生物改善葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎研究, columnId=1212692426674848354, journalTitle=中国药学杂志, columnName=炎症性肠病研究专栏, runingTitle=null, highlight=null, articleAbstract=

目的 从炎症信号通路Toll 样受体4(TLR4)/核因子κB(NF-κB)以及肠道微生物群组成改变的角度探究黄连碱衍生物Q3抗葡聚糖硫酸钠(dextran sodium sulfate, DSS)诱导小鼠溃疡性结肠炎(ulcerative colitis, UC)的作用机制。方法 将50只小鼠随机分为正常对照组、模型组、阳性药柳氮磺吡啶(SASP)组、Q3-低剂量组和Q3-高剂量组,每组10只,除正常对照组外其余组小鼠以2.5% DSS溶液喂饮建立UC模型。SASP组灌胃给予700 mg·kg^-1·d^-1SAPA,Q3低与高两个剂量组分别灌胃给予50和100 mg·kg^-1·d^-1Q3,其余组给予等量蒸馏水。给药6 d后取小鼠结肠组织进行疾病活动指数(disease activity index, DAI)评分和长度测量,苏木精-伊红染色(hematoxylin-eosin staining,HE)检测各组病理损伤程度,16S rRNA高通量测序检测小鼠肠内容物中肠道菌群的变化,免疫组织化学(IHC)检测结肠组织TLR4和p-p65的表达,Western blot法检测结肠组织中TLR4、p-p65和p-IκBα的蛋白表达,体外实验,利用脂多糖(lipopolysaccharide, LPS)诱导炎症反应,免疫荧光法检测IEC6细胞和RAW264.7细胞中NF-κB p65的核转位。结果 体内:在DSS诱导的小鼠UC模型中,与模型组相比,Q3能够显著改善UC小鼠体重降低、结肠挛缩和DAI评分升高等情况,HE染色结果显示Q3明显改善UC小鼠肠道上皮破坏、隐窝结构紊乱和杯状细胞减少等病理损伤;16S rRNA结果表明Q3可以增加DSS作用后肠道菌群的生物多样性,调节肠道菌群组成。免疫组化和Western blot结果显示Q3可以显著降低模型组小鼠结肠组织中TLR4和p-p65的表达;体外:免疫荧光和Western blot结果显示Q3能够抑制IEC6和RAW264.7两种细胞中NF-κB p65的核转位。结论 Q3能够通过抑制TLR4/NF-κB通路以及调节肠道菌群组成改善肠道炎症,发挥抗UC作用,有望成为治疗UC的候选化合物。

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Immunology, 2008, 125(2):145-153., articleTitle=The role of T‐regulatory cells and Toll‐like receptors in the pathogenesis of human inflammatory bowel disease, refAbstract=null)], funds=[Fund(id=1212786708005966661, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, awardId=2022-I2M-1-014, language=CN, fundingSource=中国医学科学院医学与健康科技创新工程资助(2022-I2M-1-014), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1212786702595314291, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, xref=null, ext=[AuthorCompanyExt(id=1212786702607897203, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, companyId=1212786702595314291, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=State Key Laboratory of Digestive Health, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China), AuthorCompanyExt(id=1212786702616285812, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, companyId=1212786702595314291, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=中国医学科学院、北京协和医学院药物研究所 消化健康全国重点实验室, 北京 100050)])], figs=[ArticleFig(id=1212786705359360774, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=EN, label=Fig.1, caption=Effects of Q3 on body weight, colon length, DAI score and pathology of UC model mice (×40). n=10,$\stackrel{-}{x}$±s

A-weight growth rate; B-colon length; C-disease activity index(DAI); D-colon morphology; E-hematoxylin-eosin staining of colons;→labeled inflammatory infiltrations and destroyed crypt structures;¹⁾ P<0.01, vs control group; ²⁾ P<0.05,³⁾ P<0.01, vs model group.

, figureFileSmall=vzQuCyt4UGIbDSDyUkFdGg==, figureFileBig=CBK5h93Xz4lf7Wu67vHX6w==, tableContent=null), ArticleFig(id=1212786705426469642, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=CN, label=图1, caption=Q3对溃疡性结肠炎(UC)模型小鼠体质量、结肠长度、疾病活动指数(DAI)评分以及病理的影响(×40)。n=10,$\stackrel{-}{x}$±s

A-体质量增长率变化;B-结肠长度;C-疾病活动指数(DAI);D-结肠形态;E-苏木精-伊红(HE)染色;→标记的是炎性浸润与被破坏的隐窝结构;与对照组相比,¹⁾ P<0.01; 与模型组相比,²⁾ P<0.05,³⁾ P<0.01。

, figureFileSmall=vzQuCyt4UGIbDSDyUkFdGg==, figureFileBig=CBK5h93Xz4lf7Wu67vHX6w==, tableContent=null), ArticleFig(id=1212786705581658896, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=EN, label=Fig.2, caption=Effects of Q3 on the expression of TLR4-related proteins in mouse colon tissue. n=10,$\stackrel{-}{x}$±s

A-immunohistochemistry of colon tissues(×200); B-expression changes of TLR4,p-p65,and p-IκBα protein in colon tissues; C-quantitative results of TLR4 expression; D-quantitative results of p-IκBα expression;¹⁾ P<0.05, vs control group;²⁾ P<0.05, vs model group.

, figureFileSmall=Kv+JLhdUcud08OXJms/r/Q==, figureFileBig=GRG/iA5DJQDszGhLd1t1LA==, tableContent=null), ArticleFig(id=1212786705682322198, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=CN, label=图2, caption=Q3对小鼠结肠组织Toll样受体4(TLR4)相关蛋白表达的影响。n=10,$\stackrel{-}{x}$±s

A-结肠组织免疫组化(×200);B-结肠组织中TLR4、p-p65、p-IκBα蛋白的表达变化;C-TLR4表达的定量结果;D-p-IκBα表达的定量结果;与对照组相比,¹⁾ P<0.05;与模型组相比,²⁾ P<0.05。

, figureFileSmall=Kv+JLhdUcud08OXJms/r/Q==, figureFileBig=GRG/iA5DJQDszGhLd1t1LA==, tableContent=null), ArticleFig(id=1212786705787179798, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=EN, label=Fig.3, caption=16S rRNA analysis of the effects of Q3 on the diversity of intestinal microbiota in mice. n=6-7

A-shannon index;B-ACE index;C-chao 1 index;D-observed-species index.

, figureFileSmall=RWfpmCtInbDwJVOwMNsV/A==, figureFileBig=YxqWgHI0eckaInb7ReBqQA==, tableContent=null), ArticleFig(id=1212786707024499480, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=CN, label=图3, caption=16S rRNA 分析Q3对小鼠肠道菌群多样性的影响。n=6~7

A-香农多样性指数;B-基于丰度的覆盖估计值指数;C-Chao 1 指数;D-观测物种指数。

, figureFileSmall=RWfpmCtInbDwJVOwMNsV/A==, figureFileBig=YxqWgHI0eckaInb7ReBqQA==, tableContent=null), ArticleFig(id=1212786707125162783, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=EN, label=Fig.4, caption=Effects of Q3 on intestinal microbiota abundance in mice at genus level. n=6-7,$\stackrel{-}{x}$±s, figureFileSmall=iS87OvELS69Mh/00RDFMbA==, figureFileBig=OcSfAVbUIQORnTMQoKL+HA==, tableContent=null), ArticleFig(id=1212786707209048870, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=CN, label=图4, caption=Q3在属水平对小鼠肠道菌群丰度的影响。n=6~7,$\stackrel{-}{x}$±s, figureFileSmall=iS87OvELS69Mh/00RDFMbA==, figureFileBig=OcSfAVbUIQORnTMQoKL+HA==, tableContent=null), ArticleFig(id=1212786707309712170, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=EN, label=Fig.5, caption=Effects of Q3 on the abundance of beneficial and pathogenic bacteria in the intestine of mice at the genus level. n=6-7,$\stackrel{-}{x}$±s

A-Effect of Q3 on the abundance of beneficial bacteria at the genus level;B-Effect of Q3 on the abundance of pathogenic bacteria at the genus level;¹⁾ P<0.05,³⁾ P<0.01, vs control group; ²⁾ P<0.05, vs model group.

, figureFileSmall=/N05wJUhPZWsyUfNKBRtZQ==, figureFileBig=71bXo7V2ZUTiEjjnK4bdpg==, tableContent=null), ArticleFig(id=1212786707393598251, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=CN, label=图5, caption=Q3在属水平对小鼠肠道有益菌群和致病菌群丰度的影响。n=6~7,$\stackrel{-}{x}$±s

A-属水平下Q3对有益菌群丰度的影响;B-属水平下Q3对致病菌群丰度的影响;与对照组相比,¹⁾ P<0.05,³⁾ P<0.01; 与模型组相比,²⁾ P<0.05。

, figureFileSmall=/N05wJUhPZWsyUfNKBRtZQ==, figureFileBig=71bXo7V2ZUTiEjjnK4bdpg==, tableContent=null), ArticleFig(id=1212786707511038768, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=EN, label=Fig.6, caption=The intestinal microbiota of mice in each group was based on the results of principal component analysis (PCA) of OTU. n=6-7,$\stackrel{-}{x}$±s, figureFileSmall=N0Cyx7RB+St1ZzUqusghGw==, figureFileBig=ql2jAkZllwYtJfMYL8iSuw==, tableContent=null), ArticleFig(id=1212786707611702072, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=CN, label=图6, caption=小鼠肠道菌群基于操作分类单元(OTU)的主成分分析(PCA)结果。n=6~7,$\stackrel{-}{x}$±s, figureFileSmall=N0Cyx7RB+St1ZzUqusghGw==, figureFileBig=ql2jAkZllwYtJfMYL8iSuw==, tableContent=null), ArticleFig(id=1212786707733336893, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=EN, label=Fig.7, caption=Effects of Q3 on changes of TLR4/NF-κB related protein expression in IEC6 and RAW 264.7 cells stimulated by LPS. n=3,$\stackrel{-}{x}$±s

A-expression of TLR4/NF-κB related proteins in IEC6 and RAW264.7 cells; B-nuclear translocation of NF-κB p65 in IEC6 and RAW264.7 cells(×63).

, figureFileSmall=VTrMkYhSuutpTBwq8IstCQ==, figureFileBig=mn+GsUj2XSKadvGo4TQZNA==, tableContent=null), ArticleFig(id=1212786707829805887, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212692427429827341, language=CN, label=图7, caption=Q3对脂多糖(LPS)刺激IEC6与RAW264.7细胞后TLR4/NF-κB相关蛋白表达变化。n=3,$\stackrel{-}{x}$±s

A-IEC6和RAW264.7细胞中TLR4/NF-κB相关蛋白表达;B-IEC6和RAW264.7细胞中NF-κB p65核转位情况 (×63)。

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