阔叶十大功劳对急性溃疡性结肠炎小鼠肠道菌群及氧化应激的影响  

Effects of Mahonia bealei(Fort.)Carr.on intestinal flora and oxidative stress in mice with acute ulcerative colitis

作　　者：智慧 龙健 涂星 胡泽华 杨宝 聂娟 刘可云 ZHI Hui;LONG Jian;TU Xing;HU Zehua;YANG Bao;NIE Juan;LIU Keyun(Health Science Center,Hubei Minzu University,Enshi 445000,China;Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu University,Enshi 445000,China;Chinese Medicinal Materials Products Quality Supevision and Inspection Center in Wuling Mountainous Area,Hubei Minzu University,Enshi 445000,China)

机构地区：[1]湖北民族大学医学部,湖北恩施445000 [2]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000 [3]湖北民族大学武陵山中药材检验检测中心,湖北恩施445000

出　　处：《中国病理生理杂志》2023年第6期1086-1094,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31860287,No.81560703);风湿性疾病发生与干预湖北省重点实验室开放基金项目(No.PT022215)。

摘　　要：目的:探讨阔叶十大功劳[Mahonia bealei(Fort.)Carr.]对葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的急性溃疡性结肠炎(ulcerative colitis,UC)小鼠肠道菌群及氧化应激的影响。方法:48只SPF级C57BL/6J小鼠,随机分为:空白组,模型组,阔叶十大功劳低、中、高剂量(2.3、4.6和9.2 g/kg生药)组,美沙拉嗪(0.2 g/kg)组。采用自由饮用2.5%DSS诱导UC小鼠模型,造模当天灌胃给药,连续10 d。苏木精-伊红(hematoxylin-eosin,HE)染色观察结肠组织并进行组织病理学评分;16S rRNA基因测序技术分析肠道菌群的变化;生化法检测小鼠肠黏膜中总超氧化物歧化酶(total superoxide dismutase,T-SOD)、还原型谷胱甘肽(reduced glutathione,GSH)和过氧化氢酶(catalase,CAT)活性及丙二醛(malondialdehyde,MDA)含量;Western blot法检测结肠组织Kelch样环氧氯丙烷相关蛋白1(Kelch-like ECH-associated protein 1,Keap1)、血红素加氧酶1(heme oxygenase-1,HO-1)、NAD(P)H醌氧化还原酶1[NAD(P)H-quinone oxidoreductase 1,NQO1]、胞浆和胞核中核因子E2相关因子(nuclear factor E2-related factor 2,Nrf2)蛋白表达。结果:与空白组相比,模型组小鼠结肠黏膜损伤显著(P<0.05),肠道菌群多样性和丰度降低,结肠抗氧化酶T-SOD、GSH及CAT活性显著降低,MDA含量显著增多(P<0.01),Keap1蛋白水平显著升高,NQO1和HO-1蛋白水平显著下降(P<0.01),胞浆Nrf2蛋白水平显著增高,胞核Nrf2蛋白水平也显著增加(P<0.01)。与模型组相比,阔叶十大功劳可修复小鼠结肠黏膜损伤,减少炎症细胞浸润(P<0.05或P<0.01),改善肠道菌群多样性及丰度,在门水平上,Firmicutes(厚壁菌门)显著上升,Bacteroidota(拟杆菌门)显著下降(P<0.05),属水平上4个菌属Muribaculaceae、Odoribacter、Rikenellaceae_RC9_gut_group(理研菌科RC9肠道群)、Lachnospirace⁃ae_UCG-001(毛螺菌科属UCG-001)可能与阔叶十大功劳对UC小鼠肠道黏膜的保护作用相关,抗氧化酶T-SOD、GSH及CAT活性显著提高(P<0.05或P<0.01),MDAIM:To explore the effects of Mahonia bealei(Fort.)Carr.on the intestinal flora and oxidative stress with acute ulcerative colitis(UC)induced by dextran sulfate sodium sulfate(DSS)in mice.METHODS:Fortyeight C57BL/6J mice were randomly divided into normal group,model group,low-dose(2.3 g/kg crude drug),mediumdose(4.6 g/kg crude drug)and high-dose(9.2 g/kg crude drug)Mahonia bealei(Fort.)Carr.groups,and mesalazine(0.2 g/kg)group.The mouse UC model was induced by drinking 2.5%DSS freely for 10 d.At the same time,drugs were chronically administered to the mice.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of colon tissues.The composition and diversity of the intestinal flora were detected by 16S rRNA gene sequencing.The activity of total superoxide dismutase(T-SOD),reduced glutathione(GSH)and catalase(CAT),and the content of malondialdehyde(MDA)were measured by biochemical detection.Western blot was used to detect the expression of Kelch-like ECHassociated protein 1(Keap1),heme oxygenase-1(HO-1),NAD(P)H-quinone oxidoreductase 1(NQO1)proteins and nuclear factor E2-related factor 2(Nrf2)in the cytoplasm and nucleus proteins in colon tissue.RESULTS:Compared with normal group,the colon mucosa were damaged severely(P<0.05),the abundance and diversity of intestinal flora were decreased,the activity of T-SOD,GSH and CAT were obviously decreased,the content of MDA was significantly increased,and Keap1 protein level was significantly increased(P<0.01)in model group.Furthermore,the protein levels of NQO1 and HO-1 were significantly decreased(P<0.01),and the protein levels of Nrf2 in cytoplasm and nucleus were significantly increased(P<0.01)in model group.Compared with model group,the pathological injury was repaired,inflammatory cell infiltration was reduced(P<0.05 or P<0.01),and the diversity and abundance of intestinal flora were regulated.At the phylum level,Firmicutes were significantly increased,while Bacteroidota were significantly decreased(P<0.05).At the genus level,four genera,Muribaculace

关 键 词：阔叶十大功劳 溃疡性结肠炎 肠道菌群 氧化应激 

分 类 号：R574[医药卫生—消化系统] R363[医药卫生—内科学]