健脾化瘀解毒方调控SIRT1/FoxO1自噬通路逆转胃癌前病变  

Jianpi Huayu Jiedu Formula Reversing Gastric Precancerous Lesions by Regulating SIRT1/FoxO1 Autophagy Pathway

作　　者：陈世龙 黎乐怡 李俊怡 甘海宁 黄雪君 赵自明 曾晓会 CHEN Shilong;LI Leyi;LI Junyi;GAN Haining;HUANG Xuejun;ZHAO Ziming;ZENG Xiaohui(The Fifth School of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou Guangdong China 510095;The Second Chinese Medicine Hospital of Guangdong Province/Guangdong Provincial Institute of Traditional Chinese Medicine Engineering and Technology,Guangzhou Guangdong China 510095;Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Research and Development,Guangzhou Guangdong China 510095)

机构地区：[1]广州中医药大学第五临床医学院,广东广州510095 [2]广东省第二中医院/广东省中医药工程技术研究院,广东广州510095 [3]广东省中医药研究开发重点实验室,广东广州510095

出　　处：《中医学报》2025年第4期858-866,共9页Acta Chinese Medicine

基　　金：广东省自然科学基金项目(2022A1515012654,2022A1515012060)。

摘　　要：目的:基于沉默信息调节因子1(sirtuin 1,SIRT1)/叉头转录因子亚家族O1(forkhead box O1,FoxO1)通路探讨健脾化瘀解毒方(JPHY)逆转胃癌前病变(gastric precancerous lesions,GPL)的作用机制。方法:将7周龄Atp4a^(-/-)小鼠随机分为Atp4a^(-/-)组、JPHY高剂量组(9 g·kg^(-1),JPHY-H)及JPHY低剂量组(4.5 g·kg^(-1),JPHY-L),以同窝野生型小鼠为空白组(WT),连续灌胃8周。采用HE染色观察胃组织病理形态,Western blot法检测胃黏膜SIRT1、FoxO1、FoxO3A、Beclin1、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、MUC2蛋白表达水平。GES-1细胞使用N-甲基-N-亚硝基脲(500μmol·L^(-1))诱导4次建立GPL模型。将细胞分成正常组(GES-1)、模型组(MTCs)、氯喹组(5μmol·L^(-1)CQ)、雷帕霉素组(50 nmol·L^(-1)Rapa)、5%含药血清组(5%JPHY)、10%含药血清组(10%JPHY)、CQ+5%JPHY组、CQ+10%JPHY组、Rapa+5%JPHY组以及Rapa+10%JPHY组。采用Transwell实验检测细胞侵袭和迁移情况,Western blot法检测细胞MUC2、CDX2、SIRT1、FoxO1、FoxO3A、Beclin1、自噬相关基因5(autophagy-related gene 5,ATG5)、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)、p-mTOR蛋白表达水平。结果:HE染色显示,WT组小鼠胃黏膜病理形态无明显异常;Atp4a^(-/-)组小鼠胃黏膜固有层可见巨型空泡样变和异型增生。与Atp4a^(-/-)组相比,JPHY-H组、JPHY-L组小鼠胃黏膜病变程度减轻。Western blot显示,与WT组比较,Atp4a^(-/-)组小鼠胃黏膜MUC2、SIRT1、FoxO1、Bcl-2蛋白表达水平升高(P<0.05);与Atp4a^(-/-)组比较,JPHY-L组小鼠胃黏膜MUC2、FoxO1、Bcl-2/Bax水平降低(P<0.05)。与GES-1组比较,MTCs组细胞迁移和侵袭数量增加(P<0.05)。与MTCs组比较,CQ组、CQ+5%JPHY组、CQ+10%JPHY组细胞迁移和侵袭数量减少(P<0.05)。与Rapa组比较,Rapa+5%JPHY组细胞迁移和侵袭数量减少(P<0.01)。与CQObjective:To investigate the mechanism of reversal of gastric precancerous lesions(GPL)by the formula of Strengthening the Spleen,Resolving Blood Stasis and Eliminating Toxins(JPHY)based on the Silent Information Regulator 1(SIRT1)/Forkhead Transcription Factor Subfamily O1(FoxO1)pathway.Methods:Seven-week-old Atp4a^(-/-)mice were randomly divided into the Atp4a^(-/-)group,the JPHY high-dose group(9 g·kg^(-1),JPHY-H)and the JPHY low-dose group(4.5 g·kg^(-1),JPHY-L),and wild-type mice from the same litter were used as the blank group(WT),and the gastric tissue was gavaged for 8 weeks.HE staining was used to observe the histopathological morphology of the stomach,and Western blot was used to detect the expression levels of SIRT1,FoxO1,FoxO3A,Beclin1,B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),MUC2,and MUC2 proteins in the gastric mucosa.GES-1 cells were induced with N-methyl-N-nitrosourea(500μmol·L^(-1))for 4 times to establish the GPL model.The cells were divided into normal group(GES-1),model group(MTCs),chloroquine group(5μmol·L^(-1)CQ),rapamycin group(50 nmol·L^(-1)Rapa),5%drug-containing serum group(5%JPHY),10%drug-containing serum group(10%JPHY),CQ+5%JPHY group,CQ+10%JPHY group,Rapa+5%JPHY group,and Rapa+10%JPHY group.Transwell assay was used to detect cell invasion and migration,and Western blot was used to detect cell MUC2,CDX2,SIRT1,FoxO1,FoxO3A,Beclin1,autophagy-related gene 5(ATG5),microtubule-associated protein 1 light chain 3(microtubule-associated protein 1 light chain 3(LC3),mammalian target of rapamycin(mTOR),and p-mTOR protein expression levels.Results:HE staining showed that there was no obvious abnormality in the pathologic morphology of the gastric mucosa of mice in the WT group;giant vacuolike changes and heterogeneous hyperplasia could be seen in the lamina propria of the gastric mucosa of mice in the Atp4a^(-/-)group.Compared with the Atp4a^(-/-)group,the degree of gastric mucosal lesions was reduced in the JPHY-H and JPHY-L groups.Western blot showed that the protein expr

关 键 词：胃癌前病变 健脾化瘀解毒方 SIRT1/FoxO1通路 自噬 

分 类 号：R285.5[医药卫生—中药学]