养阴生肌散通过AMPK/mTOR信号通路调控自噬改善克罗恩病肠纤维化的作用机制研究  

The mechanism of action of Yangyin Shengji Powder in alleviating intestinal fibrosis in Crohn disease by regulating autophagy through AMPK/mTOR signaling pathway

作　　者：戴晖 刘佳丽 刘巧玲 李昕染 卞聪慧 钱海华[1] 张丹[1] DAI Hui;LIU Jiali;LIU Qiaoling;LI Xinran;BIAN Conghui;QIAN Haihua;ZHANG Dan(Hospital of Nanjing University of Chinese Medicine,Nanjing,Jiangsu 210029,China)

机构地区：[1]南京中医药大学附属医院,江苏南京210029

出　　处：《湖南中医药大学学报》2025年第3期409-416,共8页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学青年基金项目(81904203)。

摘　　要：目的探讨养阴生肌散通过腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白(AMPK/mTOR)信号通路调控自噬改善克罗恩病大鼠肠纤维化的作用机制。方法将28只大鼠随机分为对照组、模型组、美沙拉嗪组(420 mg/kg)、养阴生肌散组(100 mg/kg),每组7只。参考Morris方法制备克罗恩病肠纤维化模型,药物组均灌肠0.5 mL,1次/d,共4周。观察大鼠的一般情况,计算疾病活动指数(DAI)评分;对结肠组织进行HE染色并行结肠黏膜损伤指数(CMDI)评分、组织学病变评分,进行Masson染色并行纤维化指数评分;采用Western blot法检测AMPK、mTOR、核糖体蛋白S6激酶(p70S6K)、真核翻译起始因子4E结合蛋白1(4EBP1)、微管相关蛋白1轻链3(LC3)蛋白水平;运用RT-PCR法检测肌球蛋白样BCL2结合蛋白(Beclin-1)、螯合体1(P62)的mRNA水平。结果与对照组比较,模型组体质量下降百分率分数显著上升(P<0.01),DAI评分、CMDI评分、组织学病变评分、纤维化指数评分均显著增高(P<0.01);LC3Ⅱ/LC3I、p-AMPK/AMPK、p-4EBP1/4EBP1、p-mTOR/mTOR表达量显著降低(P<0.01),p-p70S6K/p70S6K显著增高(P<0.01);Beclin-1表达量明显降低(P<0.01),P62表达量明显增高(P<0.01)。与模型组比较,美沙拉嗪组和养阴生肌散组体质量下降百分率分数均显著升高(P<0.01),DAI评分、CMDI评分、组织学病变评分、纤维化指数评分均显著降低(P<0.01);LC3Ⅱ/LC3I、p-AMPK/AMPK、p-4EBP1/4EBP1、p-mTOR/mTOR表达量显著升高(P<0.01),p-p70S6K/p70S6K显著降低(P<0.01);Beclin-1表达量明显上升(P<0.05),P62表达量明显下降(P<0.01)。结论养阴生肌散可能通过AMPK/mTOR通路介导的自噬激活改善克罗恩病肠纤维化。Objective To explore the mechanism of action of Yangyin Shengji Powder(YYSJP)in alleviating intestinal fibrosis in rats with Crohn disease by regulating autophagy through the adenosine monophosphate-activated protein kinase/mammalian target of rapamycin(AMPK/mTOR)signaling pathway.Methods A total of 28 rats were randomly divided into control group,model group,mesalazine group(420 mg/kg),and YYSJP group(100 mg/kg),with 7 rats in each group.The intestinal fibrosis model of Crohn disease was prepared according to the Morris method.The drug group was given enema of 0.5 mL once a day for a total of four weeks.The general condition of rats was observed,and the disease activity index(DAI)was calculated;HE staining on colon tissue was performed to evaluate the colonic mucosa damage index(CMDI)and histological lesion score.Masson staining was used to evaluate the fibrosis index;Western blot was applied to measure the protein levels of AMPK,mTOR,ribosomal protein S6 kinase(p70S6K),eukaryotic translation initiation factor 4E-binding protein 1(4EBP1),and microtubule-associated protein 1 light chain 3(LC3).RT-PCR was employed to measure the mRNA levels of Beclin-1 and sequestosome 1(P62).Results Compared with the control group,the body weight loss percentage in the model group significantly elevated(P<0.01),DAI score,CMDI score,histological lesion score,and fibrosis index score all significantly increased(P<0.01);LC3Ⅱ/LC3I,p-AMPK/AMPK,p-4EBP1/4EBP1,and p-mTOR/mTOR expression levels significantly decreased(P<0.01),while p-p70S6K/p70S6K significantly increased(P<0.01);Beclin-1 expression level significantly decreased(P<0.01),and P62 expression level significantly increased(P<0.01).Compared with the model group,the body weight loss percentage of the mesalazine group and YYSJP group both significantly increased(P<0.01).DAI score,CMDI score,histological lesion score,and fibrosis index score all significantly decreased(P<0.01);the expression levels of LC3Ⅱ/LC3I,p-AMPK/AMPK,p-4EBP1/4EBP1,and p-mTOR/mTOR all significantly increas

关 键 词：克罗恩病 肠纤维化 养阴生肌散 自噬 AMPK/mTOR信号通路 

分 类 号：R285.5[医药卫生—中药学]