湿生扁蕾呫吨酮对UC肠纤维化-CAC演变模型大鼠TGF-β1/Smad通路及EMT因子表达的影响  

Effects of Gentianopsis paludosa xanthone on the Expression of TGF-β1/Smad Pathway and EMT Factors during the Evolution of Ulcerative Colitis-related Intestinal Fibrosis to CAC in Rats

作　　者：卢年华 靳展洪叶 张永鹏[1] 张萌[1] 张海霞[1] 李俊珂 赵慧巧 靳国印[1] LU Nianhua;JIN Zhanhongye;ZHANG Yongpeng;ZHANG Meng;ZHANG Haixia;LI Junke;ZHAO Huiqiao;JIN Guoyin(College of Traditional Chinese Medicine,Hebei North University,Zhangjiakou 075000)

机构地区：[1]河北北方学院中医学院,张家口075000

出　　处：《中药药理与临床》2024年第7期63-67,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：河北省自然科学基金(编号:H2020405027、H2020405010);河北省高等学校科学技术研究项目(编号:QN2020118);河北省属高校基本科研业务项目(编号:JYT2021036、JYT2019009);河北省中医药管理局项目(编号:2020237)。

摘　　要：目的:探讨湿生扁蕾呫吨酮对大鼠溃疡性结肠炎(UC)肠纤维化向结肠炎相关性结肠癌(CAC)演变中TGF-β1/Smad通路及上皮间质转化(EMT)因子表达的影响。方法:70只Wistar大鼠随机分为正常对照组、UC肠纤维模型对照组、CAC模型对照组、湿生扁蕾呫吨酮35、70、140 mg/kg组和沙利度胺14 mg/kg组,每组10只。除正常对照组外,其余60只大鼠灌肠给予三硝基苯磺酸(TNBS)建立UC肠纤维模型,从中选取50只注射二甲苯肼(DMH)建立CAC模型,其中40只给予相应药物干预;对大鼠疾病活动指数(DAI)、结肠长度、壁厚和结肠湿质量指数等进行基础评价;Masson染色观察结肠病理变化;Western blot和qPCR法分别检测转化生长因子β1(TGF-β1)、磷酸化smad2/3(p-Smad2/3)蛋白表达和E-钙粘蛋白(Ecad)、β-连环蛋白(Bcatenin)mRNA表达,免疫组化(IHC)检测锌指蛋白Snail表达(AOD)。结果:与正常对照组比较,CAC模型对照组结肠长度缩短,壁厚和结肠湿质量指数显著升高(P<0.01),结肠内壁可见高分化腺瘤,TGF-β1和p-Smad2/3蛋白表达显著下调(P<0.01),Ecad mRNA显著下调,Bcatenin mRNA显著上调(P<0.01),Snail蛋白表达显著升高(P<0.01);与CAC模型对照组比较,湿生扁蕾呫吨酮70、140 mg/kg组结肠长度显著增加(P<0.01),壁厚、结肠湿质量指数、DAI得分显著降低(P<0.05或P<0.01),湿生扁蕾呫吨酮35、70、140 mg/kg组胶原容积百分率显著升高(P<0.01),但结肠腺瘤分化程度降低,各给药组TGF-β1和p-smad2/3蛋白表达显著上调(P<0.01),Ecad mRNA表达上调、Bcatenin mRNA表达下调,Snail蛋白表达降低(P<0.01)。结论:湿生扁蕾呫吨酮可有效抑制UC肠纤维化向CAC演变,其机制可能与促进TGF-β1/Smad通路中TGF-β1和p-Smad2/3表达,干预和调控EMT因子Ecad、Bcatenin mRNA和Snail表达有关。Objective:To investigate the effect of Gentianopsis Paludosa xanthone on the expression of transforming growth factorβ1(TGF-β1)/Smad pathway and epithelial-mesenchymal transition(EMT)factors during the evolution of ulcerative colitis(UC)-related intestinal fibrosis to colitis-associated colorectal cancer(CAC)in rats.Methods:A total of 70 Wistar rats were randomly divided into normal control group,UC-related intestinal fibrosis model group,CAC model group,Gentianopsis paludosa xanthone 35,70 and 140 mg/kg groups,and thalidomide 14 mg/kg group,with 10 rats in each group.Except for normal control group,the remaining 60 rats were given TNBS via enemator to establish a UC-related intestinal fibrosis model,and then 50 rats were injected with DMH to establish a CAC model,of which 40 rats were treated with corresponding drugs.The disease activity index(DAI),colon length,wall thickness and colon wet mass index of rats were evaluated.The pathological changes of colon were observed by Masson staining.The protein expressions of TGF-β1 and phospho-smad2/3,and mRNA levels of E-cadherin(E-cad)andβ-catenin were detected by Western blot and qPCR,respectively.The expression of Zinc finger protein Snail was detected by immunohistochemistry(IHC).Results:Compared with the normal control group,the CAC model group showed decreased colon length,increased wall thickness and colon wet mass index(P<0.01),and well-differentiated adenomas were observed in the inner wall of the colon,as well as the protein expression of TGF-β1 and P-smad2/3 were decreased(P<0.01).The mRNA level of E-cad was significantly down regulated,while the mRNA level ofβ-catenin was significantly up regulated(P<0.01).The expression of Snail was elevated(P<0.01).Compared with the CAC model group,the Gentianopsis paludosa xanthone 70 and 140 mg/kg groups showed increased colon length(P<0.01)and decreased wall thickness,colon wet mass index and DAI score(P<0.01 or P<0.05).The collagen volume fraction was increased in the Gentianopsis paludosa xanthone 35,70 and 140 m

关 键 词：湿生扁蕾呫吨酮 UC肠纤维 结肠炎相关性结肠癌 转化生长因子β1 上皮间质转化 

分 类 号：R285.5[医药卫生—中药学]