积雪草酸对葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠肠黏膜屏障的保护作用及机制研究  

Protective Effect and Mechanism of Asiatic Acid on Intestinal Mucosal Barrier Repair in Mice with DSS-Induced Ulcerative Colitis

作　　者：李阳 徐秉忠 王加玉[2] 闫梦丹 叶耘峰 LI Yang;XU Bingzhong;WANG Jiayu;YAN Mengdan;YE Yunfeng(Jiangsu College of Nursing,Huai’an 223003 Jiangsu,China;Huai’an First People’s Hospital,Huai’an 223001 Jiangsu,China)

机构地区：[1]江苏护理职业学院,江苏淮安223003 [2]淮安市第一人民医院,江苏淮安223001

出　　处：《中药新药与临床药理》2023年第5期628-635,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：淮安市“天医星”医学检验重点实验室项目(HAP202004)。

摘　　要：目的研究积雪草酸对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎(UC)小鼠肠黏膜屏障的保护作用及机制。方法将C57BL/6小鼠随机分为空白组、模型组、柳氮磺胺吡啶组(450 mg·kg^(-1))及积雪草酸低、高剂量组(3、30 mg·kg^(-1)),每组6只。小鼠自由饮用2.5%DSS溶液诱导建立UC模型,采用小鼠体质量变化及疾病活动指数(DAI)评分来评价造模效果。采用苏木素-伊红(HE)、过碘酸雪夫染色(PAS)法观察结肠组织病理变化;ELISA法检测小鼠血浆肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β水平;RT-qPCR法检测小鼠结肠组织中黏蛋白2(MUC2)、claudin-1、occludin、闭锁小带蛋白1(ZO-1)mRNA表达;Western Blot法检测小鼠结肠组织中MUC2蛋白表达;免疫组化法检测小鼠结肠组织中claudin-1、occludin、ZO-1表达。结果与空白组比较,模型组小鼠的体质量显著下降(P<0.01),DAI评分显著升高(P<0.01);模型组小鼠的结肠隐窝结构严重丧失,杯状细胞结构破坏严重;血浆TNF-α、IL-6、IL-1β水平显著升高(P<0.01);结肠组织MUC2蛋白表达及MUC2、claudin-1、occludin、ZO-1 mRNA表达显著下调(P<0.01);结肠组织中claudin-1、occludin、ZO-1阳性细胞表达显著减少(P<0.01)。与模型组比较,积雪草酸低、高剂量组小鼠的体质量显著增长(P<0.01),DAI评分显著下降(P<0.01);结肠隐窝结构恢复,炎性浸润减轻,杯状细胞结构和数量明显恢复;血浆TNF-α、IL-6、IL-1β水平明显下降(P<0.05,P<0.01);结肠组织MUC2蛋白表达及MUC2、claudin-1、occludin、ZO-1 mRNA表达明显上调(P<0.05,P<0.01);结肠组织claudin-1、occludin、ZO-1阳性细胞表达明显增加(P<0.05,P<0.01)。结论积雪草酸对DSS诱导的UC小鼠结肠组织病理损伤具有明显改善作用,可能与其上调MUC2及紧密连接蛋白的表达,保护肠黏膜屏障的完整性有关。Objective To investigate the protective effect and mechanism of asiatic acid on the intestinal mucosal barrier in mice with dextran sulfate sodium(DSS)-induced ulcerative colitis(UC).Methods C57BL/6 mice were randomly divided into blank group,model group,Salazosulfapyridine(SASP)group(450 mg·kg^(-1))and asiatic acid low-and high-dose groups(3 and 30 mg·kg^(-1)).Mice were induced to drink 2.5%DSS solution ad libitum to establish UC modeling,and changes in body mass and disease activity index(DAI)scores were used to evaluate the effect of modeling.Histopathological changes in the colon were observed by hematoxylin-eosin(HE)and periodic acid-Schiff(PAS)stain;the levels of tumor necrosis factorα(TNF-α),interleukin 6(IL-6)and IL-1βin plasma were measured by ELISA;mRNA expressions of mucin 2(MUC2),claudin-1,occludin and zonula occluden 1(ZO-1)in mouse colon tissue were detected by RT-qPCR;MUC2 protein expression in mouse colon tissue was detected by Western Blot;expressions of claudin-1,occludin,ZO-1 in mouse colon tissue were measured by immunohistochemistry.Results Compared with the blank group,the mice in the model group showed a significant decrease in body mass(P<0.01)and a significant increase in DAI score(P<0.01);the mice in the model group showed severe loss of colonic crypt structure and severe destruction of cupular cell structure;TNF-α,IL-6 and IL-1βlevels in plasma were significantly increased(P<0.01);the protein expression of MUC2 and the mRNA expressions of MUC2,claudin-1,occludin,ZO-1 were significantly down-regulated in colonic tissue(P<0.01);expressions of positive cells claudin-1,occludin,ZO-1 were significantly reduced in colonic tissue(P<0.01).Compared with the model group,mice in the low-and high-dose groups of asiatic acid showed a significant increase in body mass(P<0.01)and a significant decrease in DAI score(P<0.01);the structure of colonic crypts was restored,inflammatory infiltration was reduced,and the structure and number of cupped cells were significantly restored;plasma TNF-α,IL-6

关 键 词：积雪草酸 溃疡性结肠炎 肠黏膜屏障 黏蛋白2 紧密连接蛋白 小鼠 

分 类 号：R285.5[医药卫生—中药学]