乐胃饮加味方通过PI3K/AKT/mTOR信号通路调控糖酵解干预慢性萎缩性胃炎  

Modified Leweiyin Formula Regulates Glycolysis Through PI3K/AKT/mTOR Signaling Pathway in Treatment of Chronic Atrophic Gastritis

作　　者：叶芸 李春灵 屈孟琪 张永生[1] 陈小辉 季巾君[1] 施一春[3] 徐燕立 朱飞叶 YE Yun;LI Chun-ling;QU Meng-qi;ZHANG Yong-sheng;CHEN Xiao-hui;JI Jin-jun;SHI Yi-chun;XU Yan-li;ZHU Fei-ye(School of Basic Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053;Academy of Chinese Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053;Department of Gastroenterology,The Third Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310005;Hangzhou Chuancheng Chinese Medicine Clinic,Hangzhou 310005)

机构地区：[1]浙江中医药大学基础医学院,杭州310053 [2]浙江中医药大学中医药科学院,杭州310053 [3]浙江中医药大学附属第三医院消化内科,杭州310005 [4]杭州传承中医门诊部,杭州310005

出　　处：《中国中西医结合杂志》2025年第2期190-197,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：浙江省基础公益研究计划项目(No.LY23H270004,No.LY18H270013);2022年全国名老中医药专家传承工作室建设项目;浙江省名老中医专家传承工作室项目(No.GZS2021025)。

摘　　要：目的 基于磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(AKT)/雷帕霉素靶蛋白(mTOR)信号通路相关性,探讨乐胃饮加味方对慢性萎缩性胃炎(CAG)大鼠胃黏膜糖酵解关键调控靶点的影响。方法 采用自由饮用1-甲基-3-硝基-1-亚硝基胍(MNNG)、灌服水杨酸钠和饮食饥饱失常的复合造模法诱导CAG大鼠模型。除空白组外,造模后大鼠按体重根据随机数字表法分为模型组、维酶素组、乐胃饮加味方(中药)低剂量组(6.3 g/kg)、乐胃饮加味方(中药)高剂量组(25.2 g/kg),每组6只。正常组与模型组大鼠每日以灭菌水灌胃,各组均干预28天。采用苏木素-伊红(HE)染色法观察大鼠胃黏膜病理改变,阿利新蓝-过碘酸雪夫(AB-PAS)染色检测大鼠胃黏膜肠上皮化生状态,电子显微镜(SEM)观察大鼠胃黏膜上皮细胞表面超微结构。qRT-PCR检测各组大鼠胃黏膜组织中葡萄糖转运体1(Glut1)、己糖激酶2(HK2)、丙酮酸激酶M2亚型(PKM2)、乳酸脱氢酶A(LDHA)mRNA水平。Western Blot检测各组大鼠胃黏膜组织中Glut1、HK2、PKM2、LDHA、PI3K、AKT、mTOR蛋白表达。结果 与空白组比较,模型组大鼠体重降低(P<0.01),表现出消瘦、精神萎靡及食量减少,大鼠胃黏膜细胞发生萎缩、排列紊乱及肠上皮化生,胃黏膜细胞破损和缺失。模型组大鼠胃黏膜中Glut1、HK2、PKM2、LDHA mRNA及蛋白表达水平升高(P<0.05,P<0.01),PI3K、AKT、mTOR蛋白表达亦升高(P<0.01)。与模型组比较,各干预组大鼠体重均升高(P<0.01),饮食量、体重和精神状态均有所改善;维酶素组及中药低、高剂量组均在病理学角度修复了CAG模型大鼠的胃黏膜损伤,缓解了肠上皮化生。与模型组比较,各干预组Glut1、HK2、PKM2 mRNA水平均降低(P<0.01),且中药高剂量组LDHA mRNA水平降低(P<0.01);中药低、高剂量组PKM2、LDHA、Glut1、HK2、PI3K、mTOR蛋白表达降低(P<0.05,P<0.01),中药高剂量组AKT蛋白表达亦降低(P<0.01)。结论 乐胃饮加味方Objective To investigate the effect of modified Leweiyin Formula(MLF)on key glycolytic regulatory targets in the gastric mucosa of rats with chronic atrophic gastritis(CAG)based on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR) signaling pathway. Methods The CAG rat model was induced using a combination of free drinking of 1-methyl-3-nitro-1-nitrosoguanidine(MNNG), oral administration of sodium salicylate, and an irregular feeding schedule. After modeling, except for the blank group, the remaining rats were randomly divided into the model group, Vitacoenzyme group, Chinese medicine(CM) low-dose group(6.3 g/kg), and CM high-dose group(25.2 g/kg), with 6 rats in each group. Rats in the normal group and the model group were gavaged daily with sterilized water. Each group received daily interventions for 28 days. HE staining was used to observe pathological changes of gastric mucosa. Alisin Blue-Periodic Acid Schiff(AB-PAS)staining was used to detect the intestinal metaplasia. The changes of gastric mucosal epithelial cells were observe by Scanning electron microscopy(SEM).The mRNA expressions of glucose transporter 1(Glut1), hexokinase 2(HK2), pyruvate kinase M2(PKM2), and lactate dehydrogenase A(LDHA)were detected using qRT-PCR. The protein expressions of Glut1, HK2, PKM2, LDHA, PI3K, AKT, and mTOR were detected using Western Blot. Results Compared with the blank group, the model group showed decreased body weight(P<0.01), reduced food intake, lethargy, and pathological changes in the gastric mucosa, including atrophy and intestinal metaplasia. mRNA and protein expressions of Glut1, HK2, PKM2, and LDHA increased(P<0.05, P<0.01), the protein expressions of PI3K, AKT, and mTOR increased in the model group(P<0.01). Compared with the model group, the body weight of rats in all intervention groups increased(P<0.01), and their food intake, body weight, and mental state were all improved;the Vitacoenzyme group and the CM low-dose group, as well as the CM high-dose group,

关 键 词：乐胃饮加味方 慢性萎缩性胃炎 糖酵解 维酶素 磷脂酰肌醇-3激酶/蛋白激酶B/雷帕霉素靶蛋白 中药复方 中西医结合 

分 类 号：R73[医药卫生—肿瘤]