马甲子干预实验性结肠炎相关结直肠癌前期病变的网络药理学及实验研究  

Network Pharmacology and Animal Experiment of Paliurus ramosissimus Intervention in Experimental Colitis-Associated Colorectal Precancerous Lesions

作　　者：赵欢 胡竟一[2] 白筱璐[2] 雷玲[2] 吴瑕[2] 林雪燕 牛圆圆 陈雨慧 余悦[2] 李东晓[2] 徐超群[2] ZHAO Huan;HU Jingyi;BAI Xiaolu;LEI Ling;WU Xia;LIN Xueyan;NIU Yuanyuan;CHEN Yuhui;YU Yue;LI Dongxiao;XU Chaoqun(School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 610041;Sichuan Provincial Key Laboratory of Chinese Materia Medica Quality and Innovative Chinese Medicine Research/Sichuan-Chongqing Joint Key Laboratory for the Creation and Quality Evaluation of New Chinese Medicines,Sichuan Academy of Chinese Medicine Sciences,Chengdu 610041)

机构地区：[1]成都中医药大学药学院,成都610041 [2]四川省中医药科学院,中药材品质及创新中药研究四川省重点实验室/中药新药创制与质量评价川渝共建重点实验室,成都610041

出　　处：《中药药理与临床》2024年第3期65-74,共10页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家重大新药创制项目(编号:2018ZX09731013);四川省中医药人才培养项目;四川省中药药理学重点学科建设项目(编号:2020ZDXK01);四川省科技厅省级科研院所基本科研业务专项项目(编号:2022JDKY0013)。

摘　　要：目的:研究马甲子(Paliurus ramosissimus,PR)对结肠炎相关结直肠癌(colitis-associated colorectal cancer,CAC)前期病变的干预作用,探索作用机制。方法:采用基因功能注解(GO)、基因数据库(KEGG)富集分析和分子对接等方法,分析马甲子提取物中主要药效成分白桦脂酸干预CAC前期病变的潜在靶点和通路;采用腹腔注射氧化偶氮甲烷(Azoxymethane,AOM)及经饮水摄入葡聚糖硫酸钠(Dextran sulfate sodium,DSS)对小鼠进行循环处理方式,建立实验性CAC前期病变小鼠模型,连续42 d灌胃给予马甲子提取物205、820 mg/kg,观察其对小鼠模型疾病活动指数(DAI)、结直肠组织病理等的影响,ELISA法检测血清中癌胚抗原(CEA)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量,q RT-PCR法检测结直肠组织中信号转导和转录激活子3(Stat3)m RNA表达。结果:网络药理学研究筛选出157个白桦脂酸干预CAC的潜在作用靶点,对激素的反应、跨膜受体蛋白酪氨酸激酶信号通路、细胞迁移的正向调节、等874个生物过程,激酶结合、蛋白激酶活性等75个分子功能,膜筏、谷氨酸能突触等37个细胞组分,涉及的主要通路有癌症通路、癌症中的蛋白聚糖、脂质和动脉粥样硬化、前列腺癌、JAK-STAT信号通路;白桦脂酸与IL-1β、STAT3等靶点亲和力均<-5 kcal/mol。与正常对照组比较,模型对照组小鼠死亡率明显增高(P<0.05),脾脏指数及组织病理评分显著升高(P<0.01),异型增生等提示癌前病变,血清中CEA、IL-1β、TNF-α含量显著升高(P<0.01),Stat3 m RNA的表达显著上调(P<0.01);与模型对照组比较,马甲子提取物820 mg/kg组组织病理评分明显降低(P<0.05),血清中CEA、IL-1β、TNF-α含量显著降低(P<0.01),Stat3 m RNA的表达显著下调(P<0.01)。结论:马甲子提取物经口给药可抑制AOM/DSS所致实验性CAC癌前病变,其作用机制与炎症抑制及下调STAT3等有关。Objective:To investigate the interventional effects of Paliurus ramosissimus(PR)on colitis-associated colorectal precancerous lesions and explore the underlying mechanism.Methods:Gene Ontology(GO)annotation,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,and molecular docking were employed to predict the potential targets and pathways of betulinic acid(the main pharmacological constituent of PR extract)on colitis-associated colorectal precancerous lesions.The mouse model of experimental colitis-associated colorectal precancerous lesions was established by intraperitoneal injection of azoxymethane(AOM)and intake of dextran sulfate sodium(DSS)via drinking water in a circulating manner.After administration of PR extract at 820 mg/kg and 205 mg/kg by gavage for 42 consecutive days,the disease activity index(DAI)and colorectal histopathology of the modeled mice were observed.ELISA was performed to measure the levels of carcinoembryonic antigen(CEA),interleukin(IL)-1β,and tumor necrosis factor(TNF)-αin blood.Quantitative realtime polymerase chain reaction(qRT-PCR)was employed to determine the mRNA level of signal transducer and activator of transcription 3(Stat3)in the colorectal tissue.Results:A total of 157 potential targets of betulinic acid for intervention in colitis-associated colorectal precancerous lesions were predicted.The targets were annotated to 874 biological processes(e.g.,responses to hormones,protein tyrosine kinase signaling pathway,and positive regulation of cell migration),75 molecular functions(e.g.,binding to kinase and protein kinase activity),and 37 cell components(e.g.,membrane raft and glutamatergic synapse).The targets were mainly enriched in the cancer pathway,proteoglycan in cancer,lipid and atherosclerosis,prostate cancer,JAK-STAT signaling pathway,etc.Betulinic acid showed the binding energy less than-5 kcal/mol with IL-1β,STAT3,and other targets.Compared with the normal control group,the model control group showed increased mortality(P<0.05),elevated spleen index and

关 键 词：马甲子 白桦脂酸 结肠炎相关结直肠癌 网络药理学 分子对接 癌胚抗原 信号转导和转录激活子3 

分 类 号：R285.5[医药卫生—中药学]