马勃大黄栓对小鼠溃疡性结肠炎的治疗作用及机制研究  

Therapeutic effect of Mabo Dahuang suppository on ulcerative colitis in mice and related mechanism

作　　者：郭子霞 张楠 邢媛 李炜[1] 张丹参[4] GUO Zi-xia;ZHANG Nan;XING Yuan;Li Wei;ZHANG Dan-shen(Department of Pharmacy,Hebei North University,Zhangjiakou Hebei 075000;Central Laboratory,Hebei Key Laboratory of Brain Sciences and Psychiatric-Psychologic Diseases,The First Hospital of Hebei Medical University,Shijiazhuang 050031;Department of Neurology,Brain Aging and Cognitive Neuroscience Laboratory of Hebei,The First Hospital of Hebei Medical University,Shijiazhuang 050031;School of Chemical and Pharmaceutical Engineering,Hebei University of Science and Technology,Shijiazhuang 050018)

机构地区：[1]河北北方学院药学系,河北张家口075000 [2]河北医科大学第一医院中心实验室、河北省脑科学与精神心理疾病重点实验室,石家庄050031 [3]河北医科大学第一医院神经内科、河北省脑老化与认知神经科学实验室,石家庄050031 [4]河北科技大学化学与制药工程学院,石家庄050018

出　　处：《中南药学》2022年第12期2701-2707,共7页Central South Pharmacy

基　　金：国家自然科学基金青年项目(No.82100863);河北省重点研发计划项目生物医药专项(No.20372509D);河北省自然科学基金(No.H2020208032,No.H2020206105,No.H2020206643);河北省中医药类科研计划课题(No.2020268)。

摘　　要：目的建立溃疡性结肠炎小鼠模型并探究马勃大黄栓对该模型的治疗作用及机制。方法联用噁唑酮(OXZ)和2,4,6-三硝基苯磺酸(TNBS)建立溃疡性结肠炎小鼠模型。将造模小鼠随机分为模型组,柳氮磺胺吡啶栓组,马勃栓组,大黄栓组,马勃大黄栓高、中、低剂量(5 mg-2.5 mg、2.5 mg-1.25 mg、1.25 mg-0.625 mg)组,正常饲养的小鼠为正常对照组。各组栓剂给药14 d。通过记录给药期间各组小鼠疾病活动指数(DAI)、观察结肠黏膜损伤并评分(CMDI)、HE染色观察结肠组织病理学变化并评分(HPS)、检测结肠组织中的髓过氧化物酶(MPO)活力,评价马勃大黄栓对小鼠溃疡性结肠炎的治疗作用;通过ELISA法检测血清IL-1β及IL-6水平、免疫组织化学染色观察结肠TNF-α蛋白表达情况、免疫组织化学染色及Western blot法观察测定结肠TLR4、MyD88、NF-κB蛋白表达情况,探究马勃大黄栓治疗溃疡性结肠炎的作用机制。结果与正常对照组小鼠相比,模型组小鼠DAI、结肠单位长度湿重、CMDI及HPS评分均显著升高(P<0.01)。给药14 d后各给药组小鼠与模型组相比,其症状及结肠损伤得到不同程度缓解,DAI、CMDI及HPS、MPO活力均降低(P<0.05或P<0.01)。与单用马勃、大黄治疗相比,马勃大黄栓高剂量组小鼠DAI、CMDI、HPS、MPO活力、血清IL-1β、IL-6及结肠中TNF-α、TLR4、MyD88、NF-κB蛋白表达水平均降低(P<0.05或P<0.01)。结论联用OXZ和TNBS建立的溃疡性结肠炎小鼠模型具有稳定、迁延性特点;马勃、大黄联用明显改善溃疡性结肠炎小鼠症状、减轻结肠损伤,其抗炎作用机制可能与抑制TLR4/MyD88/NF-κB通路有关。Objective To establish a mouse model of ulcerative colitis and determine the therapeutic effect of Mabo Dahuang suppository on the model and its mechanism.Methods Ulcerative colitis mouse model was induced by combining oxazolone with 2,4,6-trinitrobenzene sulfonic acid.The model mice were randomly divided into a model group,a sulfasalazine suppository group,a Mabo suppository group,a Dahuang suppository group,and Mabo Dahuang suppository three dose groups(5 mg-2.5 mg,2.5 mg-1.25 mg,and 1.25 mg-0.625 mg).The mice fed normally served as the normal control group.The mice were given suppository continuously for 14 days.The therapeutic effect of Mabo Dahuang suppository on ulcerative colitis mice was evaluated with recorded disease activity index(DAI)during the administration,scores of the colon mucosa damage index(CMDI),histopathological changes of colon by HE staining,and the activity of myeloperoxidase(MPO)in the colon tissue.The serum IL-1βand IL-6 levels were detected by ELISA.TNF-αprotein expression in the colon was observed by immunohistochemical staining,the protein expression of TLR4,MyD88 and NF-κB in the colon was determined by immunohistochemical staining and Western blot.Results Compared with the normal control group,the DAI,colon weight/length ratio,CMDI and HPS in the model group were significantly increased(P<0.01).After 14 days of administration,compared with the model group,the symptoms and colon damage of the mice in each administration group were improved to different degrees,and the DAI,CMDI,HPS and MPO activity were decreased(P<0.05 or P<0.01).Compared with the use of Lasiosphaera seu Calvatia or Rhei Radix et Rhizoma alone,the DAI,CMDI,HPS and MPO activity,the serum IL-1βand IL-6 levels and the protein expression of TNF-α,TLR4,MyD88 and NF-κB in the colon of mice high dose in the combination suppository group were all decreased(P<0.05 or P<0.01).Conclusion The mouse model of ulcerative colitis induced by combining OXZ and TNBS is stable and persistent.Lasiosphaera seu Calvatia combined wit

关 键 词：马勃 大黄 溃疡性结肠炎 炎症 TLR4/MyD88/NF-κB通路 

分 类 号：R285[医药卫生—中药学]