基于YAP/TAZ信号通路探讨白术内酯I对UC大鼠肠屏障功能障碍的影响  

Base on YAP/TAZ Signaling Pathway to Investigate the Effect of Atractylenolide I on Intestinal Barrier Dysfunction in UC Rats

作　　者：李森 田利晖 韩涛 闫玲新 王素梅 李宏坤 LI Sen;TIAN Lihui;HAN Tao(Handan Central Hospital,Hebei Handan 056000,China)

机构地区：[1]河北省邯郸市中心医院,河北邯郸056000 [2]河北医科大学第二医院中医科,河北石家庄050000

出　　处：《河北医学》2024年第12期1993-1999,共7页Hebei Medicine

基　　金：河北省中医药管理局2024年度中医药类科学研究课题计划项目,(编号:2024194)。

摘　　要：目的:探讨白术内酯I(Atr-I)对溃疡性结肠炎(UC)大鼠肠屏障功能障碍及Yes相关蛋白(YAP)/含有PDZ结合基序的转录共激活因子(TAZ)信号通路的影响。方法:构建UC大鼠模型,将所有实验大鼠分为UC组、17.5、35.0mg/kg Atr-I剂量组(Atr-I-L、Atr-I-H组)、35.0mg/kg Atr-I+17.5mg/kg Yap抑制剂维替泊芬组(Atr-I-H+VTPF组),每组18只,另外取18只大鼠作为对照组(Control组);实验结束后测量各组小鼠体重并进行疾病活动指数(DAI)评分;测量结肠长度并进行苏木精-伊红(HE)染色观察各组大鼠结肠病理学形态变化;酶联免疫吸附法(ELISA)检测血清白细胞介素(IL)-1β、IL-17、γ干扰素(IFN-γ);免疫组化检测各组大鼠结肠组织肠黏膜屏障相关蛋白带状闭合蛋白1(ZO-1)、闭合蛋白1(Claudin-1)蛋白阳性表达情况;蛋白质印迹法(Western blot)法检测结肠组织YAP、p-YAP、TAZ蛋白表达水平。结果:与Control组比较,UC组大鼠结肠黏膜上皮细胞排列紊乱,杯状细胞明显减少,大量炎性细胞浸润,部分区域有溃疡形成,甚至增厚黏连、肠管变形,黏膜充血、水肿明显,大鼠体重、结肠长度、ZO-1、Claudin-1阳性表达、p-YAP/YAP、TAZ表达显著降低,DAI评分、IL-1β、IL-17、IFN-γ水平显著升高(P<0.05);与UC组相比,Atr-I-L、Atr-I-H组大鼠结肠黏膜上皮损伤有所改善,大鼠体重、结肠长度、ZO-1、Claudin-1阳性表达、p-YAP/YAP、TAZ表达依次显著升高,DAI评分、IL-1β、IL-17、IFN-γ水平依次显著降低(P<0.05);与Atr-I-H组相比,Atr-I-H+VTPF组大鼠结肠黏膜上皮损伤加重,大鼠体重、结肠长度、ZO-1、Claudin-1阳性表达、p-YAP/YAP、TAZ表达显著降低,DAI评分、IL-1β、IL-17、IFN-γ水平显著升高(P<0.05)。结论:Atr-I可降低DAI评分,抑制炎症反应,改善结肠黏膜上皮损伤,改善UC大鼠的肠屏障功能障碍,其作用机制可能与激活YAP/TAZ信号通路有关。Objective:To investigate the effects of atractylenolide I(Atr-I)on intestinal barrier dysfunction and Yes-associated protein(YAP)/transcription coactivator containing PDZ binding motif(TAZ)signaling pathway in rats with ulcerative colitis(UC).Methods:UC rat model was constructed,all experimental rats were divided into UC group,17.5,35.0 mg/kg Atr-I dose group(Atr-I-L,Atr-I-H group),35.0 mg/kg Atr-I+17.5 mg/kg Yap inhibitor vitipofin group(Atr-I-H+VTPF group),18 rats in each group,and 18 rats were taken as control group(Control group).After the experiment,the body weight of mice of each group was measured and the disease activity index(DAI)was calculated after intervention.The length of the colon was measured and the pathological morphology changes of the colojisun in each group were observed by hematoxylin-eosin(HE)staining.Serum interleukin(IL)-1β,IL-17 and interferon-γ(IFN-γ)were detected with enzyme-linked immunosorbent assay(ELISA).The positive expression of intestinal mucosal barrier-related protein band-closing protein 1(ZO-1)and claudin-1(Claudin-1)in colon tissue of rats of each group were detected by immunohistochemistry.The protein expression levels of YAP,p-YAP and TAZ in colon tissues were detected by western blotting(Western blot).Results:Compared to Control group,the colonic mucosal epithelial cells in UC group were disordered,the goblet cells were significantly reduced,a large amount of infiltration inflammatory cells were found,and ulcers were formed in some areas,even thickening and adhesion,intestinal deformation,mucosal congestion and edema were obvious,body weight,colon length,ZO-1,Claudin-1 positive expression,p-YAP/YAP,and TAZ expression were significantly decreased,and the DAI score,IL-1β,IL-17,and IFN-γlevels were obviously increased(P<0.05).Compared with UC group,the colonic mucosal epithelial injury of rats in Atr-I-L and Atr-I-H groups was improved,body weight,colon length,positive expression of ZO-1,Claudin-1,p-YAP/YAP and TAZ were significantly increased in turn,and the DAI score

关 键 词：溃疡性结肠炎 白术内酯I YAP/TAZ信号通路 肠屏障功能障碍 

分 类 号：R285.5[医药卫生—中药学]