文献类型：期刊文献

中文题名：乌梅丸调控Keap-1-Nrf2/HO-1信号通路抑制溃疡性结肠炎小鼠氧化应激损伤

英文题名：Wumeiwan regulate Keap-1-Nrf2/HO-1 signaling pathway to inhibit oxidative stress injury in mice with ulcerative colitis

作者：杜丽东[1,2];王颖[1];辛蕊华[3];仇正英[3];赵冠宇[1];李能莲[1];邵晶[1,2];吴国泰[1,2]

第一作者：杜丽东

机构：[1]甘肃中医药大学,药学院,甘肃兰州730000;[2]甘肃中医药大学,陇药产业创新研究院,甘肃兰州730000;[3]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州730050

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2024

卷号：40

期号：14

起止页码：2088

中文期刊名：中国临床药理学杂志

外文期刊名：The Chinese Journal of Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：甘肃省自然科学基金资助项目(20JR10RA317);甘肃省中医药科研立项课题基金资助项目(GZK-2019-23);甘肃省中医药研究中心开放课题基金资助项目(ZYZX-2020-20)。

语种：中文

中文关键词：乌梅丸;溃疡性结肠炎;Kelch-样ECH相关蛋白1-核因子E2相关因子2/血红素加氧酶1信号通路;氧化应激损伤

外文关键词：Wumeiwan;ulcerative colitis;Kelch-like ECH associated protein 1-nuclear factor E2 related factor 2/heme oxygenase-1 signaling pathway;oxidative stress injury

摘要：目的研究乌梅丸调控Kelch样ECH相关蛋白1(Keap-1)-核因子E2相关因子2(Nrf2)/血红素加氧酶1(HO-1)信号通路对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎(UC)小鼠氧化应激损伤的抑制作用。方法将40只C57BL/6J小鼠随机分为正常组、模型组、阳性对照组、低剂量实验组和高剂量实验组,每组8只。自由饮用2%DSS水溶液的方法复制UC模型。造模同时,正常组和模型组小鼠灌胃等体积0.9%NaCl,阳性对照组小鼠灌胃美沙拉嗪(5-ASA)溶液0.005 g·10 g^(-1)·d-1,低、高剂量实验组小鼠分别灌胃乌梅丸水煎液0.13、0.26 g·10 g^(-1)·d-1,持续7 d。评价小鼠疾病活动指数(DAI)和结肠长度变化;用实时荧光定量聚合酶链反应检测小鼠结肠组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、环氧酶2(COX-2)、诱导型一氧化氮合酶(iNOS)mRNA表达水平;用蛋白质印迹法检测小鼠结肠Kelch样ECH相关蛋白(Keap-1)、核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)蛋白的表达水平。结果正常组、模型组、阳性对照组和低、高剂量实验组第7天的DAI分别为0、2.62±0.33、1.87±0.35、1.87±0.35和1.58±0.35,结肠长度分别为(8.16±0.47)、(5.98±0.24)、(7.58±0.38)、(7.33±0.24)和(7.48±0.51)cm,SOD mRNA表达水平分别为1.01±0.16、0.40±0.01、1.43±0.45、0.65±0.01和0.83±0.02,CAT mRNA表达水平分别为1.01±0.20、0.45±0.01、0.84±0.02、0.68±0.07和0.87±0.05,COX-2 mRNA表达水平分别为1.03±0.33、16.65±0.60、4.78±0.25、14.07±0.60和7.39±0.15,iNOS mRNA表达水平分别为1.04±0.40、20.71±0.66、8.09±0.93、15.44±0.68和11.66±0.06,Keap-1蛋白表达水平分别为1.22±0.16、1.10±0.05、1.18±0.05、1.94±0.08和1.17±0.08,Nrf2蛋白表达水平分别为1.12±0.16、0.76±0.15、0.65±0.13、0.70±0.16和0.82±0.18,HO-1蛋白表达水平分别为1.34±0.15、1.00±0.12、0.89±0.10、1.50±0.18和1.40±0.13。模型组的上述指标与正常组比较,低、高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(P<0.01,P<0.05)。结论乌梅丸对溃疡性结肠炎小鼠具有抗氧化应激损伤作用,其作用机制可能与调节结肠Keap-1-Nrf2/HO-1信号通路蛋白表达有关。
Objective To investigate the inhibitory effects of Wumeiwan on oxidative stress injury of ulcerative colitis mice induced by dextran sulfate sodium(DSS)by regulating Kelch-like ECH related protein 1(Keap-1)-nuclear factor E2 related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathwayand.Methods Forty C57BL/6 mice were randomly divided into five groups:normal group,model group,positive control group,experimental-L,-H groups.UC mice model were induced by free access to 2%DSS water.Mice in normal and model group were orally administered with 0.9%NaCl,mice in positive control group were orally treated with Mesalazine solution(0.005 g·10 g^(-1)·d^(-1)),while mice in experimental groups were orally administered with Wumeiwan decoction at the dose of 0.13 and 0.26 g·10 g^(-1)·d^(-1),respectively.All the drugs were administered for consecutive 7 days,1 times a day.The levels of disease activity index(DAI)and the colon length were scored.The levels of superoxide dismutase(SOD),catalase(CAT),cyclooxygenase-2(COX-2)and inducible nitric oxide synthase(iN OS)in colon tissue of mice were determined by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)method.The level of Keap-1,Nrf2,HO-1 proteins in colon tissue were determined by Western blot method.Results The levels of DAI of seventh day in normal group,positive control group,experimental-L,-H groups were 0、(2.62±0.33),(1.87±0.35),(1.87±0.35)and(1.58±0.35);the colon lengths were(8.16±0.47)、(5.98±0.24),(7.58±0.38),(7.33±0.24)and(7.48±0.51)cm;the SOD mRNA were 1.01±0.16、0.40±0.01,1.43±0.45,0.65±0.01 and 0.83±0.02;the CAT mRNA were1.01±0.20、0.45±0.01,0.84±0.02,0.68±0.07 and 0.87±0.05;the COX-2 mRNA were 1.03±0.33、16.65±0.60,4.78±0.25,14.07±0.60 and 7.39±0.15;the iN OS mRNA were 1.04±0.40、20.71±0.66,8.09±0.93,15.44±0.68 and 11.66±0.06;the levels of Keap-1 were 1.22±0.16、1.10±0.05,1.18±0.05,1.94±0.08 and 1.17±0.08;the levels of Nrf2 were 1.12±0.16、0.76±0.15,0.65±0.13,0.70±0.16 and0.82±0.18;the levels of HO-1 were 1.34±0.15、1.00±0.12,0.89±0.10,1.50±0.18 and 1.40±0.13,respectively.Significant difference was found between normal group and model group(P<0.01,P<0.05);significant difference was also found between the experimental-L,-H groups and model group(P<0.01,P<0.05).Conclusion Wumeiwan can inhibit oxidative stress in mice with UC,the mechanisms may be related to adjusted the expression of Keap-1-Nrf2/HO-1 signaling pathway protein in colon.