黄芩汤治疗慢性复发型溃疡性结肠炎大肠湿热证小鼠的作用机制研究  

Study on the Mechanism of Huangqin Decoction on Chronically Recurrent Ulcerative Colitis in Mice with Damp-Heat Syndrome of Large Intestine

作　　者：吴玉竹 李敏瑶 邱建国 雷骏轩 孙泽佳 濮声乐 王小林[5] 郑学宝 黄晓其[1,2] WU Yuzhu;LI Minyao;QIU Jianguo;LEI Junxuan;SUN Zejia;PU Shengle;WANG Xiaolin;ZHENG Xuebao;HUANG Xiaoqi(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Dongguan Institute of Guangzhou University of Chinese Medicine,Dongguan 523808 Guangdong,China;Guangdong University of Technology,Guangzhou 510006 Guangdong,China;Zhejiang Normal University,Jinhua 321004 Zhejiang,China;Beijing Institute of Nanoenergy and Nanosystems,,Chinese Academy of Sciences,Beijing 101400,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006 [2]东莞广州中医药大学研究院,广东东莞523808 [3]广东工业大学,广东广州510006 [4]浙江师范大学,浙江金华321004 [5]中国科学院北京纳米能源研究所,北京101400

出　　处：《中药新药与临床药理》2023年第8期1083-1094,共12页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金资助项目(82274227);广东省自然科学基金资助项目(20231800936162,2022A1515140011,2021B1515140043,2022A1515010092);广州中医药大学“双一流”与高水平大学学科协同创新团队培育项目(2021xk34)。

摘　　要：目的探讨黄芩汤在慢性复发型溃疡性结肠炎大肠湿热证(UCM)中清热利湿的作用机理。方法将40只小鼠随机分为正常组、模型组及黄芩汤低、中、高剂量组。模型组小鼠使用人工气候箱、白酒、高脂饲料及不同浓度的葡聚糖硫酸钠(DSS)溶液联合诱导;黄芩汤组在造模第35天至第44天给予不同剂量黄芩汤干扰;实验期间,观察小鼠一般情况,包含体质量、饮食、粪便等,并进行湿热评分、疾病活动指数(DAI)评分;通过课题组研发的My color软件检测爪尾红色值占比变化;HE染色观察结肠及舌组织病理变化;检测小鼠血清中超氧化物歧化酶(SOD)和髓过氧化物酶(MPO)含量;Western Blot法检测结肠组织中水通道蛋白及NF-κB信号通路相关蛋白表达量;ELISA法检测干扰素γ(IFN-γ)、脂多糖(LPS)、巨噬细胞迁移抑制因子(MIF)、血栓素B2(TXB2)含量;qRT-PCR法检测结肠组织中白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和Toll样受体4(TLR4)表达水平。结果与正常组比较,模型组体质量降低,DAI评分、湿热评分、爪尾红色值占比增加;舌组织丝状乳头紊乱、角质层增厚,结肠组织受损严重,长度缩短;结肠组织TXB2、IFN-γ含量、血清MIF含量增加,LPS水平增加;血清SOD活性降低、MPO活性增加;IL-6、IL-1β、TNF-α mRNA水平上升;结肠AQP1、AQP3、AQP8蛋白表达降低;IκB磷酸化增加,核外p65表达减少,核内p65表达增加,TLR4 mRNA水平上升,差异均有统计学意义(P<0.05,P<0.01)。与模型组比较,黄芩汤高剂量组体质量下降幅度减少,DAI评分、湿热评分降低,爪尾图像红色比例下降,结肠长度增加,病理情况改善,血清中的MPO、MIF、LPS水平降低,SOD含量增加,结肠组织中的TXB2、IFN-γ水平减少,IL-6、IL-1β、TNF-αmRNA水平下调,结肠AQP1、AQP3、AQP8蛋白表达增高,TLR4 mRNA水平下降,IκB磷酸化减少,核外p65表达增加,核内p65表达减少,差异均有统Objective To investigate the mechanism of Huangqin Decoction(HQD)on chronically recurrent ulcerative colitis(UC)in mice with damp-heat syndrome of large intestine.Methods Forty mice were randomly divided into 5 groups,including normal group,model group,HQD low-,medium-and high-dose groups.The model mice were established by using artificial climate chamber and feeding with white wine,high fat diet and different concentrations of dextran sulfate sodium(DSS).From the days 35 to 44 of modeling,different doses of HQD were given to interfere with the mice,and the general conditions of the mice were observed,including body mass,diet,feces,etc.The damp-heat scores and disease activity index(DAI)scores were recorded.The change in the proportion of red value of paw and tail was detected by My color software,which was developed by our research group.The pathological changes of colon and tongue were observed by HE staining.The contents of superoxide dismutase(SOD)and myeloperoxidase(MPO)in serum of mice were detected.The expression levels of aquaporins and NF-κB signaling pathway-related proteins in colon tissues were detected by Western Blot.Interferon(IFN-γ),lipopolysaccharide(LPS),macrophage migration inhibitor factor(MIF)and thromboxane B2(TXB2)were determined by ELISA.The expression levels of interleukin-6(IL-6),tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β)and Toll-like receptor 4(TLR4)in colon tissues were determined by qRT-PCR.Results Compared with the normal group,the body mass of the model group was decreased,DAI scores,damp-heat scores and the proportion of red value of paw and tail were increased.The filamentous papillae in tongue tissue were disorder,cuticle was thickened,colon tissue was seriously damaged and its length was shortened.The contents of TXB2,IFN-γ in colon tissue and serum MIF were increased,and the LPS level was increased.Serum SOD activity decreased,and serum MPO activity increased.The mRNA levels of IL-6,IL-1β and TNF-α were increased.The protein expressions of AQP1,AQP3,AQP8 in co

关 键 词：黄芩汤 溃疡性结肠炎 大肠湿热证 慢性复发型 清热利湿 肠道水液紊乱 小鼠 

分 类 号：R285.5[医药卫生—中药学]