摘要
目的研究黄芩汤对溃疡性结肠炎(UC)的治疗作用,并通过TLR2/MyD88/NF-κB信号通路探讨其治疗机制。方法将小鼠随机分为空白组、模型组、阳性对照组(柳氮磺胺吡啶,0.45 g·kg^(-1))、黄芩汤组(9.1 g·kg^(-1)),自由饮用2.5%葡聚糖硫酸钠(DSS)10 d造模,同时灌胃给药。观察小鼠体质量、结肠长度等体征变化,并进行疾病活动指数评分(DAI)。采用酶联免疫法检测血清细胞因子肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和白细胞介素1β(IL-1β)水平;HE和PAS染色法分析结肠组织病理变化;Western Blot法检测结肠组织Toll样受体2(TLR2)、髓样分化因子88(MyD88)、核转录因子-κB p65(NF-κB p65)和IL-6蛋白表达;免疫组化法检测结肠组织中TLR2、MyD88、NF-κB p65和IL-6阳性细胞表达。结果与空白组比较,模型组小鼠体质量明显下降(P<0.01),DAI评分明显升高(P<0.01),结肠明显缩短(P<0.01),血清TNF-α、IL-6和IL-1β水平明显升高(P<0.01),结肠隐窝结构严重丧失,黏膜下层水肿明显,并伴大量炎症浸润,杯状细胞结构破坏严重,数量大量减少,结肠组织中TLR2、MyD88、NF-κB p65和IL-6蛋白表达明显升高(P<0.01),结肠组织中TLR2、MyD88、NF-κB p65和IL-6阳性细胞数及表达强度明显升高(P<0.01)。与模型组比较,黄芩汤组小鼠体质量、DAI评分、结肠长度及血清细胞因子水平明显改善(P<0.01),结肠组织病变、杯状细胞结构和数量均得到恢复,结肠组织中TLR2、MyD88、NF-κB p65和IL-6蛋白表达强度和阳性细胞数及表达强度明显降低(P<0.01)。结论黄芩汤可能通过抑制TLR2/MyD88/NF-κB信号通路抑制溃疡性结肠炎小鼠的炎症,实现治疗作用。
Objective To investigate the effect of Huangqin Decoction on mice with ulcerative colitis(UC)based on TLR2/MyD88/NF-κB signaling pathway.Methods Mice were randomly divided into control group,model group,positive drug group(salazosulfapyridine,0.45 g·kg-1),Huangqin Decoction group(9.1 g·kg-1)and given 2.5%dextran sulfate sodium(DSS)freely for 10 days to induce UC.At the same time the drug was administered by oral administration.The changes of body mass of mice,disease activity index(DAI)and colon length were observed.The levels of TNF-α,IL-6 and IL-1βin serum were detected by ELISA,the pathological changes of colonic tissue were analyzed by HE and PAS staining,the expressions of TLR2,MyD88,NF-κB p65 and IL-6 in colonic tissue were detected by Western Blot and immunohistochemical staining.Results Compared with the control group,the body weight of mice dropped significantly(P<0.01),DAI score of mice increased significantly(P<0.01),the colon length of mice was shortened significantly(P<0.01)and the levels of TNF-α,IL-6 and IL-1βin serum increased significantly(P<0.01)in the model group.Severe loss of colonic crypt and obvious edema of submucosal,accompanied by massive inflammatory infiltration,serious damages of goblet-cell structures and significant reduction of the number of goblet cells were found.The protein expressions of TLR2,MyD88,NF-κB p65 and IL-6 in colon tissue increased significantly(P<0.01).The number of positive cells and expression intensity of TLR2,MyD88,NF-κB p65 and IL-6 in colon tissue were significantly increased(P<0.01).Compared with the model group,the body weight,DAI score,colon length and serum cytokine levels of mice in the Huangqin Decoction group were significantly improved(P<0.01).Tissue lesions in colon,the structure and number of goblet cells in the Huangqin Decoction group were all recovered;the expression intensity of TLR2,MyD88,NF-κB p65 and IL-6 proteins as well as the number of positive cells in the Huangqin Decoction group were significantly reduced(P<0.01).Conclusion Therapeutic effect of Huangqin Decoction on against inflammation in UC mice may be capable of inhibiting TLR2/MyD88/NF-κB signaling pathway.
作者
严宝飞
许晨新
陈灵
刘圣金
王迎
刘嘉
YAN Baofei;XU Chenxin;CHEN Ling;LIU Shengjin;WANG Ying;LIU Jia(Jiangsu Health Vocational College,Nanjing 211800 Jiangsu,China;Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine,and Key Laboratory of Chinese Medicinal Resources Recycling Utilization,State Administration of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing 210023 Jiangsu,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2022年第6期727-735,共9页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
江苏省高校“青蓝工程”优秀教学团队培养对象项目(苏教师[2020]42号)
江苏省中医药科技发展计划项目(YB2020100)
江苏卫生健康职业学院校级课题重点项目(JKB202003)。
