补脾益肠丸抑制树突状细胞炎性分化及促进紧密连接蛋白表达治疗溃疡性结肠炎的作用机制  

Mechanism of Bupi Yichang Pills alleviating ulcerative colitis by suppressing inflammatory differentiation of dendritic cells and promoting expression of tight junction proteins

作　　者：肖秋萍 钟友宝 黄佳琦 陈丽玲[1] 李姗姗[1] 施旻[1] 刘端勇[1,2] 刘荣华 XIAO Qiuping;ZHONG Youbao;HUANG Jiaqi;CHEN Liling;LI Shanshan;SHI Min;LIU Duanyong;LIU Ronghua(Jiangxi University of Chinese Medicine,Nanchang 330004,China;Institute of Traditional Chinese Medicine Health Industry,China Academy of Chinese Medical Sciences,Nanchang 330115,China)

机构地区：[1]江西中医药大学,南昌330004 [2]中国中医科学院中医药健康产业研究所,南昌330115

出　　处：《中华中医药杂志》2024年第5期2204-2210,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82260863);江西省自然科学基金项目(No.20232BAB206172);江西省卫生健康委员会科技计划(No.202110121);江西省中医药管理局科技计划(No.2022A344);江西省教育厅科学技术研究项目(No.GJJ201261)。

摘　　要：目的:研究补脾益肠丸调节树突状细胞(DCs)炎性分化及紧密连接蛋白(TJs)表达缓解溃疡性结肠炎(UC)的作用机制。方法:低、中、高剂量补脾益肠丸及5-氨基水杨酸(5-ASA)连续14 d治疗2.5%葡聚糖硫酸钠(DSS)诱导的UC模型小鼠,每天称重并观察粪便黏稠度及便血情况,采用疾病活动指数(DAI)评估结肠炎症状,实验结束后,取结肠称重并测量长度,HE染色评价结肠组织损伤情况,流式细胞术、ELISA法分别检测脾脏不同表型DCs及结肠炎性因子水平,免疫组化、q PCR检测结肠TJs的蛋白及m RNA表达水平。结果:中、高剂量补脾益肠丸能显著减轻结肠炎小鼠腹泻、便血症状,并显著降低小鼠DAI、结肠质量及结肠病理组织损伤(P<0.05),抑制DCs炎性分化,使脾脏炎性DCs(CD11c^(+)CD103^(-)E-cadherin^(+)、CD11c^(+)CD103^(-)CD80^(+)、CD11c^(+)CD103^(-)MHC-Ⅱ^(+)、CD11c^(+)CD103^(-)CD86^(+)、CD11c^(+)CD103^(-)TNF-α^(+)DCs)细胞比例显著下调(P<0.01),炎性因子(IFN-γ、IL-1β、IL-6及IL-23)水平亦显著降低(P<0.05,P<0.01);此外,中、高剂量补脾益肠丸均能显著上调Caludin1、Occludin及ZO-1的蛋白及m RNA表达水平(P<0.05,P<0.01)。结论:补脾益肠丸能有效缓解UC,其作用与抑制DCs炎性分化及促进TJs表达紧密相关。Objective:The mechanism of Bupi Yichang Pills alleviating ulcerative colitis(UC)were explored by inflammatory differentiation of dendritic cells(DCs)and expression of tight junction proteins(TJs).Methods:Mice in a UC model induced by 2.5%dextran sulfate sodium(DSS)were treated with low,medium,and high doses of Bupi Yichang Pills and5-aminosalicylic acid(5-ASA)via gavage for 14 consecutive days.They were weighed daily,and the consistency of feces and presence of blood in the stool were observed.Disease activity index(DAI)was used to assess colitis symptoms.At the end of the experiment,the colon was weighed and its length measured.HE staining was used to evaluate colonic tissue damage.Flow cytometry and ELISA were employed to detect different phenotypes of DCs in the spleen and levels of colonic inflammatory factors,respectively.Immunohistochemistry and q PCR were utilized to measure the protein and m RNA expression levels of colonic tight junctions(TJs).Results:High and medium doses of Bupi Yichang Pills significantly alleviated diarrhea and bloody stool symptoms in mice with colitis,and significantly reduced DAI,colon weight,and colonic histopathological damage(P<0.05).They inhibited the inflammatory differentiation of DCs,leading to a significant decrease in the proportion of inflammatory DCs in spleen(CD11c^(+)CD103^(-)E-cadherin^(+),CD11c^(+)CD103^(-)CD80^(+),CD11c^(+)CD103^(-)MHC-Ⅱ^(+),CD11c^(+)CD103^(-)CD86^(+),CD11c^(+)CD103^(-)TNF-α^(+)DCs)(P<0.01),as well as a significant decrease in levels of inflammatory cytokines(IFN-γ,IL-1β,IL-6,and IL-23)(P<0.05,P<0.01).Furthermore,both medium and high doses of Bupi Yichang Pills significantly upregulated the protein and m RNA expression levels of Claudin1,Occludin,and ZO-1(P<0.05,P<0.01).Conclusion:Bupi Yichang Pills effectively alleviates UC,which is closely related to inhibiting the inflammatory differentiation of DCs and promoting the expression of TJs.

关 键 词：补脾益肠丸 溃疡性结肠炎 炎性树突状细胞 紧密连接蛋白 作用机制 

分 类 号：R285.5[医药卫生—中药学]