中药复方益糖康对db/db小鼠小肠肠道屏障损伤的修复作用及机制探讨  

Repair Effect and Mechanism of Traditional Chinese Medicine Compound Yitangkang(益糖康)on Intestinal Barrier Damage in db/db Mice

作　　者：陈胡蓉 安继仁 杨宇峰[1] 刘军彤 石岩[1] CHEN Hurong;AN Jiren;YANG Yufeng;LIU Juntong;SHI Yan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Hebei University of Traditional Chinese Medicine,Shijiazhuang 050200,Hebei,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]河北中医药大学,河北石家庄050200

出　　处：《辽宁中医药大学学报》2024年第7期31-36,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家重点基础研究发展计划(973计划)项目(2013CB532004);辽宁省教育厅高校基本科研项目(JYTZD2023199);青年岐黄学者项目(2022年);辽宁省科学技术应用基础研究项目(2022020244-JH2/1013);辽宁省糖尿病中医病证结合重点实验室项目(2019JH6/10200009)。

摘　　要：目的观察中药复方益糖康对db/db小鼠小肠炎症的抑制作用及小肠黏膜损伤的修复作用并探讨其作用机制。方法将24只db/db小鼠随机分为模型组(db/db组)、益糖康组(YTK组)以及阳性药(选用利拉鲁肽Liraglutide)对照组(Liraglutide组),另选择8只C57BL/6J小鼠为对照组(Con组)。YTK组小鼠予以益糖康煎剂灌胃30 g·kg^(-1)·d^(-1),Liraglutide组每日腹腔注射利拉鲁肽200μg·kg^(-1)·d^(-1),Con组和db/db组给予等量蒸馏水灌胃,给药时间共计5周。药物处理5周后,对小鼠禁食12 h,取小鼠尾尖血进行血糖检测,之后进行动物取材和组织分离。用ELISA检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)表达;免疫荧光法检测Occludin、F4/80、CD86、CD206表达;Western blot检测法检测小肠Occludin、ZO-1、TNF-α蛋白表达。结果与Con组相比,db/db组小鼠血糖明显升高(P<0.05),与db/db组相比,Liraglutide组与YTK组血糖明显下降(P<0.05)。与db/db组相比,Liraglutide组与YTK组Occludin和ZO-1表达较明显增强(P<0.05);Western blot结果显示Occludin蛋白表达与免疫荧光趋势一致(P<0.05);与Con组相比,db/db组小鼠血清炎症因子IL-6与IL-1β均明显升高(P<0.05),Liraglutide组与YTK组较db/db组明显下降(P<0.05);与Con组比较,db/db小鼠小肠黏膜及黏膜下层IL-1β、IL-6表达量明显增强(P<0.05),而Liraglutide组与YTK组呈中度表达,较db/db组明显减弱(P<0.05)。western blot结果显示,TNF-α表达与免疫荧光趋势一致;Liraglutide组与YTK组F4/80的表达较db/db组明显减弱(P<0.05);CD86和CD206在Con组小鼠小肠表达较弱;在db/db组表达明显增强(P<0.05),其中CD86表达更为显著,而Liraglutide组与YTK组明显改善(P<0.05)。结论中药复方益糖康可以下调F4/80、CD86和CD206等巨噬细胞相关蛋白的表达,缓解小肠巨噬细胞聚集和活化,下调小肠IL-Iβ、IL-6和TNF-α等炎症因子,上调肠通道蛋白Occludin和ZO-1表达,修复db/db小鼠小肠黏膜损伤。Objective To observe the inhibitory effect of traditional Chinese medicine Compound Yitangkang(益糖康)on small intestinal inflammation and the repair effect of small intestinal mucosal damage in db/db mice,and explore its mechanism of action.Methods 24 db/db mice were randomly divided into model group(db/db group),Yitangkang group(YTK group),and positive drug group control group(Liraglutide group).Additionally,8 C57BL/6J mice were selected as the control group(Con group).YTK group mice were given Yitangkang decoction by gavage of 30 g·kg^(-1)·d^(-1),while Liraglutide group mice were given daily intraperitoneal injection of 200μg·kg^(-1)·d^(-1),Con group and db/db group were given equal amount of distilled water by gavage for a total of 5 weeks.After 5 weeks of drug treatment,the mice were fasted for 12 hours,and their tail tip blood was taken for blood glucose testing.Subsequently,animal samples and tissue isolation were performed.ELISA was used to detect the expression of IL-1βand IL-6;Immunofluorescence assay was used to detect the expression of Occludin,F4/80,CD86,and CD206;Western blot detection method for detecting small intestine Occludin,ZO-1,and TNF-αprotein expression.Results Compared with the Con group,the blood glucose levels in the db/db group were significantly increased(P<0.05),while the blood glucose levels in the Liraglutide group and YTK group were significantly decreased compared with db/db group(P<0.05).Compared with the db/db group,the expression of Occludin and ZO-1 were significantly increased in the Liraglutide group and YTK group(P<0.05);Western blot results showed that the expression of Occludin protein was consistent with the trend of immunofluorescence(P<0.05);Compared with the Con group,the expression levels of IL-1βand IL-6 in the small intestine mucosa and submucosa of db/db mice were significantly increased(P<0.05),while the Liraglutide group and YTK group showed moderate expression,which was significantly reduced compared with the db/db group(P<0.05).Western blot results

关 键 词：糖尿病 小肠炎症 巨噬细胞 益糖康 肠道屏障 

分 类 号：R285.5[医药卫生—中药学]