基于网络药理学及实验验证探究葛根素治疗溃疡性结肠炎的作用机制  

Application of network pharmacology and experimental validation in investigating therapeutic potential of puerarin for ulcerative colitis

作　　者：但文利 赵鑫 鲁星妤 郭紫婵 秦琪 李娟 唐康 张慧媛 施京红 陈丽华 DAN Wenli;ZHAO Xin;LU Xingyu;GUO Zichan;QIN Qi;LI Juan;TANG Kang;ZHANG Huiyuan;SHI Jinghong;CHEN Lihua(College of Medicine Technology,Shaanxi University of Traditional Chinese Medicine,Xianyang 712046,China;School of Basic Medical Science,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]陕西中医药大学医学技术学院,咸阳712046 [2]空军军医大学基础医学院,西安710032

出　　处：《中国免疫学杂志》2024年第5期1055-1063,共9页Chinese Journal of Immunology

基　　金：国家自然科学基金面上项目(82071848)。

摘　　要：目的:探索葛根素(PUE)对溃疡性结肠炎(UC)的治疗效果及作用机制。方法:利用网络药理学和分子对接技术对PUE调控UC的靶点进行筛选和分析。C57BL/6小鼠自由饮用2.5%DSS水溶液7 d后,观测PUE对小鼠体质量变化及疾病活动指数(DAI)评分的影响。HE染色观察小鼠结肠组织病理变化;阿利新蓝染色观察小鼠结肠组织杯状细胞变化;qRT-PCR和ELISA检测小鼠结肠组织炎症细胞因子表达。CCK-8检测PUE对MODE-K细胞活力的影响;流式细胞术检测PUE对MODE-K细胞凋亡的影响。结果:筛选得到AKT1、TNF、STAT3、CASP3、HIF1A等38个PUE调节UC的靶点,主要涉及TNF、IL-17和PI3K-Akt等信号通路。体内实验结果显示,与DSS组相比,DSS+PUE组小鼠结肠缩短程度、体质量减轻程度和DAI评分降低,结肠组织炎症细胞浸润减少,结肠组织TNF-α、IL-1βmRNA和蛋白表达降低。体外实验证实PUE减轻DSS诱导的上皮细胞凋亡。结论:PUE可减轻DSS诱导的小鼠UC发生发展。Objective:To explore therapeutic efficacy and mechanism of puerarin(PUE)in treating of ulcerative colitis(UC).Methods:Network pharmacology and molecular docking technique were used to screen and analyze targets of PUE in regulating UC.C57BL/6 mice were given free access to 2.5%DSS aqueous solution for 7 days,and influence of PUE on changes in body weight and disease activity index(DAI)score were subsequently observed.Histopathological alterations of colon tissue were observed by HE staining,changes of goblet cell population in colon tissue were evaluated through Alcian blue staining;expressions of inflammatory factors in colon tissue were detected by qRT-PCR and ELISA.Effect of PUE on MODE-K cell viability and apoptosis were assessed by CCK-8 and flow cytometry.Results:A total of 38 common targets of PUE in modulating UC,such as AKT1,TNF,STAT3,CASP3,HIF1A and etc,mainly involving TNF,IL-17 and PI3K-Akt signaling pathway.In vivo experiments confirmed that PUE ameliorated degree of colon shortening,body weight and DAI scores and reduced inflammatory cell infiltration in mice.Besides,expressions of inflammatory factors in colon,such as TNF-αand IL-1β,were inhibited by PUE.Furthermore,in vitro experiments validated that PUE relieved DSS-induced apoptosis of epithelial cells.Conclusion:PUE alleviates occurrence and development of DSS-induced UC in mice.

关 键 词：葛根素 溃疡性结肠炎 网络药理学 上皮细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]