苦豆子总碱对溃疡性结肠炎小鼠结肠及肝组织中FXR、TGR5、CYP7A1蛋白表达的影响  

Effect of total alkaloids from Sophora alopecurides on FXR,TGR5,CYP7A1 protein expression in colon and liver tissues of mice with ulcerative colitis

作　　者：肖莹 华永丽[1] 贾娅倩 董嘉琪 李芳 魏彦明[1] 袁子文[1] 纪鹏[1] XIAO Ying;HUA Yong-li;JIA Ya-qian;DONG Jia-qi;LI Fang;WEI Yan-ming;YUAN Zi-wen;Ji Peng(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070

出　　处：《天然产物研究与开发》2021年第9期1452-1462,共11页Natural Product Research and Development

基　　金：甘肃省高等学校创新基金(2020A-057);国家自然科学基金(31860717);甘肃农业大学伏羲杰出人才培育计划(Gaufx-03J01);财政部和农业农村部:国家现代农业产业技术体系资助(CARS-37)。

摘　　要：基于法尼醇X受体和G蛋白偶联胆汁酸受体5(farnesoid X receptor/G protein-coupled bile acid receptor 5,FXR/TGR5)通路探讨苦豆子总碱对小鼠溃疡性结肠炎(ulcerative colitis,UC)模型的治疗作用及其作用机制。将60只健康雄性BALB/c小鼠分空白对照组(Con)、葡聚糖硫酸钠模型组(DSS)、柳氮磺胺嘧啶阳性对照组(SASP)和苦豆子总碱治疗组(TASA)。给予BALB/c小鼠3.5%DSS饮水使其连续自由饮用7天以建立小鼠急性UC模型,造模的同时每天灌胃苦豆子总碱(75 mg/kg)进行治疗,并以柳氮磺胺嘧啶(450 mg/kg)为阳性对照药物。试验期间对各组小鼠临床症状进行疾病活动指数(disease activity index,DAI)评分。第8天观察结肠HE染色病理切片,糖原PAS染色以及阿利新蓝染色观察结肠组织杯状细胞。酶联免疫吸附法(ELISA)检测结肠组织匀浆上清液IL-10、IL-23、IL-1β及IL-17的表达。蛋白质免疫印迹(Western blot)检测小鼠结肠和肝组织FXR、TGR5、CYP7A1的表达。结果显示,经苦豆子总碱治疗后,小鼠疾病活动指数、结肠组织病理学损伤及结肠长度明显改善(P<0.01),结肠组织杯状细胞数量显著增加(P<0.01),细胞因子IL-10显著升高(P<0.01);IL-23、IL-17、IL-1β显著降低(P<0.01),血清胆汁酸含量显著升高(P<0.01),结肠和肝脏组织FXR、TGR5、CYP7A1均不同程度上调。结果表明苦豆子总碱对DSS诱导的小鼠溃疡性结肠炎有一定的治疗作用,其主要通过抑制相关炎症因子来抑制结肠炎症的发展,以及上调FXR/TGR5信号通路的相关蛋白的表达发挥其治疗作用。Based on FXR/TGR5(farnesoid X receptor/G protein-coupled bile acid receptor 5)pathway to explore the therapeutic effect and mechanism of action of total alkaloids from Sophora alopecuroides on 3.5%dextran sodium sulfate induced ulcerative colitis model in mice.Sixty healthy male BALB/c mice were divided into blank control group(Con),dextran sodium sulfate model group(DSS),sulfasalazine positive control group(SASP)and total alkaloids from S.alopecuroides treatment group(TASA).BALB/c mice were given 3.5%DSS water for continuous free drinking for 7 days to establish the mouse model of acute UC.During the modeling,S.alopecuroides extract(75 mg/kg)was given daily for treatment,and sulfasalazine(450 mg/kg)was used as the positive control drug.Clinical symptoms of mice in each group were scored by DAI during the experiment.Day eight,the pathological sections of colon were observed by HE staining,glycogen PAS staining and Alcian blue staining were used to observe the expression of goblet cells in colon tissue.The expressions of IL-10,IL-23,IL-1βand IL-17 in serum and colon tissue homogenate supernatant were detected by ELISA.The expressions of FXR,TGR5 and CYP7A1 in liver and colon tissues of mice were detected by Western blot.The results showed that after total alkaloids from S.alopecuroides extract treatment,the DAI,histopathological damage of colon and the length of colon were significantly improvement(P<0.01),the number of goblet cells in colon tissue was significantly increased(P<0.01),anti-inflammatory factor IL-10 was significantly increased(P<0.01),pro-inflammatory factors IL-23,IL-17 and IL-1βwere significantly decreased(P<0.01),serum bile acid content was significantly increased(P<0.01),FXR,TGR5 and CYP7A1 in colon and liver were up-regulated.Results showed that the total alkaloids from S.alopecuroides had a certain therapeutic effect on DSS inducing UC in mice.It inhibited the development of colitis mainly by inhibiting the related inflammatory factors,and up-regulating the expression levels of related prote

关 键 词：苦豆子总碱 UC模型 FXR TGR5 CYP7A1 

分 类 号：R932[医药卫生—生药学]