靛花通过抑制NLRP3炎症小体和保护肠道屏障治疗溃疡性结肠炎的作用机制  

Purified Indigo naturalis alleviated ulcerative colitis by protecting colon barrier and inhibiting NLRP3 inflammasome pathway

作　　者：马乐乐 刘雪梅 黄胜杰 徐超 袁小玉 阳向波 杨明 韩丽[1] 许润春[1] 张定堃[1,4] MA Le-le;LIU Xue-mei;HUANG Sheng-jie;XU Chao;YUAN Xiao-yu;YANG Xiang-bo;YANG Ming;HAN Li;XU Run-chun;ZHANG Ding-kun(State Key Laboratory of Southwestern Chinese Medicine Resources,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Sichuan Liancheng Xunkang Pharmaceutical Co.,Ltd,Yaan 625000,China;Key Laboratory of Modern Preparation of Traditional Chinese Medicine,Ministry of Education,Jiangxi University of Traditional Chinese Medicine,Nanchang 330000,China;Sichuan Provincial Engineering Research Center of Innovative Re-development of Famous Classical Formulas,Tianfu TCM Innovation Harbour,Chengdu University of Traditional Chinese Medicine,Chengdu 611930,China)

机构地区：[1]成都中医药大学药学院,西南特色中药资源国家重点实验室,四川成都611137 [2]四川联成迅康医药股份有限公司,四川雅安625000 [3]江西中医药大学现代中药制剂教育部重点实验室,江西南昌330000 [4]成都中医药大学,天府中医药创新港四川省经典名方二次创新开发工程研究中心,四川成都611930

出　　处：《药学学报》2025年第4期965-975,共11页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金项目面上项目(82173976);国家重点研发计划中医药现代化研究(2018YFC1707205);江西中医药大学全国重点实验室项目(2023QJZDSYS003)。

摘　　要：青黛作为一种传统中药,在治疗溃疡性结肠炎方面具有显著疗效。现代生产工艺将打靛形成的沉淀物——“粗靛”,除杂干燥后制得青黛成品,已成为现代药用的主流形式。“靛花”取打靛形成的顶层泡沫状物的干燥品,其中靛蓝和靛玉红的含量高于粗靛。本研究评估了粗靛和靛花在治疗溃疡性结肠炎中的疗效并探讨其作用机制。使用3%葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导急性结肠炎小鼠模型(实验获得成都中医药大学动物伦理委员会批准,批准号:2024075)。同时,小鼠灌胃给药粗靛(400、200和100 mg·kg^(-1))和靛花(400、200、100和50 mg·kg^(-1))溶液。通过酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)检测结肠组织中的髓过氧化物酶(myeloperoxidase,MPO)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6和IL-18水平来评估粗靛和靛花的抗炎活性。采用蛋白免疫印迹和免疫荧光法评估紧密连接蛋白-1(zonula occludens1,ZO-1)和膜整合蛋白(occludin)的表达。此外,还采用蛋白免疫印迹和实时荧光定量PCR(RT-q PCR)分析结肠组织中腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)、NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)和相关因子的表达水平。结果显示,粗靛和靛花可有效缓解DSS诱导的溃疡性结肠炎。与粗靛相比,靛花在治疗溃疡性结肠炎中效果更显著,尤其是在靛花200 mg·kg^(-1)剂量下。本研究进一步证实了粗靛和靛花具有显著抗炎和保护肠道屏障完整性的能力。此外,粗靛和靛花治疗溃疡性结肠炎作用可以通过抑制AMPK/SIRT1通路介导的NLRP3炎症小体的形成和激活来阐明。Indigo naturalis[Baphicacanthus cusia(Nees)Bremek.,QD],as a traditional Chinese medicine,has exhibited efficacy in the ulcerative colitis(UC).Cu Dian(CD),the current form of Indigo naturalis,has been regarded as the mainstream form of medicine today.Dian Hua(DH)is the traditional purified form of QD,in which the content of indigo and indirubin is higher than that of CD.The study evaluated the efficacy of DH and CD in UC and explored their mechanism.Male BALB/c mice were subjected to an 8-day regimen of 3%dextran sodium sulphate(DSS)drinking water to induce UC.The experiment was approved by the Animal Ethics Committee of Chengdu University of Traditional Chinese Medicine(approval number:2024075).Concurrently,the mice received intragastric administration of CD(400,200,and 100 mg·kg^(-1))and DH(400,200,100,and 50 mg·kg^(-1))for the same duration.The anti-inflammatory properties of CD and DH were evaluated by quantifying levels of myeloperoxidase(MPO),tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6,and IL-18 in colon tissue.Western blot and immunofluorescence assays were employed to assess the protein levels of zonula occludens 1(ZO-1)and occludin.Additionally,Western blot and RT-q PCR were utilized to analyze the protein and gene expression levels of AMP-activated protein kinase(AMPK),nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3),and related factors in colon tissue.CD and DH demonstrated a capacity to alleviate inflammatory responses in mice with UC.The protective impact of both CD and DH on the intestinal mucosal barrier was associated with an elevation of ZO-1 and occludin.Furthermore,the antiinflammatory effects of CD and DH were attributed to the inhibition of the NLRP3 inflammasome through the activation of the AMPK/silent information regulator of transcription 1(SIRT1)pathway.Notably,DH exhibited a more pronounced improvement in UC compared to CD,particularly at the dosage of DH-M(200 mg·kg^(-1)).Our investigation substantiates the effectiveness of

关 键 词：青黛 靛花 溃疡性结肠炎 肠道屏障 AMPK/SIRT1 NLRP3 

分 类 号：R967[医药卫生—药理学]