黄蜀葵根多糖治疗慢传输型便秘作用与网络药理学机制研究  

Pharmacodynamics of Polysaccharides from Abelmoschus Manihot Radix in Treating Slow Transit Constipation and Mechanistic Study of Network Pharmacology

作　　者：徐文丽 李舟媛 王梓灵 陈乐[2] 陈科力 侯晓莹 刘大会 杜鸿志 XU Wenli;LI Zhouyuan;WANG Ziling;CHEN Le;CHEN Keli;HOU Xiaoying;LIU Dahui;DUHongzhi(Resource Center for Chinese Materia Medica,Hubei University of Chinese Medicine,Wuhan 430065,China;Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,National and Local CollaborativeEngineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine,Key Laboratory ofChinese Medicinal Resources Recycling Utilization,National Administration of Traditional Chinese Medicine,NanjingUniversity of Chinese Medicine,Nanjing 210023,China;Wuhan Institutes of Biomedical Sciences,School of Medicine,Jianghan University,Wuhan 430056,China)

机构地区：[1]湖北中医药大学药学院中药资源与中药化学湖北省重点实验室,武汉430065 [2]南京中医药大学、江苏省中药资源产业化过程协同创新中心、中药资源产业化与方剂创新药物国家地方联合工程研究中心、国家中医药管理局中药资源循环利用重点研究室,南京210023 [3]江汉大学医学部武汉生物医学研究院,武汉430056

出　　处：《世界科学技术-中医药现代化》2024年第12期3071-3085,共15页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家财政部和农业农村部国家现代农业产业技术体系项目(CARS-21):黄冈综合试验站;负责人:刘大会;中华中医药学会青年人才托举工程(2021-QNRC2-B16);负责人:杜鸿志。

摘　　要：目的首先分析黄蜀葵根多糖的单糖组成,并进行治疗慢传输型便秘的作用评价,最后通过网络药理学、分子对接技术探讨黄蜀葵根多糖中改善慢传输型便秘的药效物质和分子机制。方法水提醇沉法制备黄蜀葵根多糖,通过离子色谱法测定黄蜀葵根多糖的单糖组成;采用盐酸洛哌丁胺(10 mg·kg^(-1))诱导的慢传输型便秘模型,以粪便含水率和小肠推进率两个指标评价其治疗便秘的治疗作用;通过苏木素-伊红(HE)染色法和免疫组化法检测结肠组织病理变化以及闭锁小带蛋白-1(ZO-l)蛋白和紧密连接蛋白claudin-1表达;利用RT-qPCR法检测各组结肠组织AQP3、AQP4、TNF-α、IL-1β和IL-6 mRNA表达水平,最后通过网络药理学和分子对接探究黄蜀葵根多糖治疗STC的潜在核心靶点及相关信号通路,探索其分子机制。结果黄蜀葵根多糖能增加小鼠粪便含水率及小肠推进率(P<0.01),抑制AQP3、AQP4、TNF-α、IL-1β和IL-6mRNA的表达(P<0.01,P<0.001)。同时,黄蜀葵根多糖高剂量组可保护结肠屏障的完整性并显著上调便秘小鼠结肠组织ZO-1和claudin-1蛋白表达。此外,网络药理学结果显示,鼠李糖、岩藻糖和葡萄糖醛酸主要作用于STAT3、JUN、CASP3、HSP90AA1、VEGFA和IL-1β等关键靶点,通过调节TRP通道的炎症介质调节、EGFR酪氨酸激酶抑制剂耐药性和糖尿病并发症中的AGE-RAGE信号通路改善便秘。Western blot结果显示黄蜀葵根多糖干预可显著降低便秘小鼠结肠组织的CASP3、VEGFA和IL-1β3蛋白表达量。结论黄蜀葵根多糖可通过多成分、多靶点、多途径治疗慢传输型便秘,为后续的临床应用和药物开发提供科学依据(国家专利已授权ZL202310894613.3),有望促进黄蜀葵非药用部位的资源高效利用。Objective The monosaccharide composition of Abelmoschus Manihot Radix polysaccharide was identified and then we evaluated the therapeutic effect on slow transit constipation.Finally,the pharmacodynamic substances and molecular mechanisms in the polysaccharide from Abelmoschus Manihot Radix to improve slow transit constipation were explored by network pharmacology and molecular docking techniques.Methods The polysaccharide from Abelmoschus Manihot Radix has been prepared by aqueous-alcoholic precipitation and has been determined by HPAEC method;The mice model of slow transit constipation was made by sc loperamide(10 mg·kg^(-1))and the therapeutic effect for the treatment of constipation was evaluated by two indicators:fecal water content and small intestinal propulsion rate;Pathological changes in the colon tissue of STC mice were observed by HE.Immunohistochemical method was used to detect zonula occludens-1(ZO-1)and claudin-1 expression in colon tissue of STC mice;qRT-PCR method was used to detect mRNA expressions of AQP3,AQP4,TNF-α,IL-1β and IL-6 in each group;Network pharmacology and molecular docking technology were used to explore the potential targets and pathways of the polysaccharide from Abelmoschus Manihot Radix in treating slow transit constipation.Results The Polysaccharide from Abelmoschus Manihot Radix significantly increased fecal water content and intestinal propulsion rate in mice caused by slow transit constipation,decreased the expressions of AQP3,AQP4,TNF-α,IL-1β and IL-6 mRNA(P<0.01,P<0.001),protected the integrity of the colonic barrier in STC mice,and increased the protein expressions of ZO-1 and claudin-1 in colon tissues of STC mice.By network pharmacology,it was found that monosaccharides such as rhamnose,fucose and glucuronic acid could mainly act on key targets such as STAT3,JUN,CASP3,HSP90AA1,VEGFA and IL-1βand regulate the Inflammatory mediator regulation of TRP channels,EGFR tyrosine kinase inhibitor resistance and AGERAGE signaling pathway in diabetic complications to improve

关 键 词：黄蜀葵 非药用部位 多糖 慢传输型便秘 网络药理学 

分 类 号：R285[医药卫生—中药学]