灭幽汤对幽门螺杆菌相关性胃炎脾胃湿热证小鼠PTEN-PI3K-Akt-FoxO凋亡信号通路的影响  

Effect of Mieyou Decoction on PTEN-PI3K-Akt-FoxO apoptotic signal pathway in the H.Pylori-associated gastritis mice with spleen-stomach damp-heat syndrome

作　　者：曾蓉 喻斌[2] 徐寅[2] 周义方 陈末 ZENG Rong;YU Bin;XU Yin;ZHOU Yifang;CHEN Mo(Hunan University of Traditional Chinese Medicine,Changsha 410208,Hunan,China;The First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410208,Hunan,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410208

出　　处：《现代中西医结合杂志》2020年第26期2869-2875,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：湖南省教育厅科研计划项目(2016C1237);湖南省临床医疗技术创新引导项目(2018SK51201);湖南省自然科学青年基金项目(2019JJ50463);湖南省研究生科研创新项目(CX20190593);湖南中医药大学中医学国内一流建设学科(2018-2020年)。

摘　　要：目的观察灭幽汤对幽门螺杆菌相关性胃炎脾胃湿热证小鼠胃组织中PTEN、PI3K、p-Akt、Akt、FoxO3a蛋白及mRNA表达的影响,探讨灭幽汤治疗脾胃湿热证可能的作用机制。方法将60只BALB/c小鼠随机分为对照组、模型组、灭幽汤低剂量组、灭幽汤中剂量组、灭幽汤高剂量组、胃四联组,每组10只。除对照组外,其余组均采用复合病因方法造模。造模成功后,对照组和模型组小鼠给予蒸馏水灌胃,1次/d;灭幽汤低、中、高剂量组小鼠分别给予灭幽汤6.2 g/kg、12.4 g/kg、24.8 g/kg灌胃,1次/d;胃四联组小鼠给予泮托拉唑肠溶胶囊+枸橼酸铋钾片+替硝唑片+克拉霉素片280.6 mg/kg灌胃,早晚各1次。各组均连续灌胃14 d,观察各组小鼠一般情况。灌胃结束后,HE染色观察胃黏膜组织病理学表现,TUNEL法检测胃黏膜细胞凋亡情况,采用Western blot法检测胃组织中PTEN、PI3K、p-Akt、FoxO3a蛋白表达情况,采用RT-PCR法检测胃组织中PTEN、PI3K、Akt、FoxO3a mRNA表达情况。结果模型组小鼠出现懒动、食欲下降、皮毛松弛无光泽、体质量减轻、肛温升高,胃黏膜组织HE染色表现为慢性浅表性胃炎;胃四联组、灭幽汤中剂量组、灭幽汤高剂量组小鼠给予相应药物灌胃后各种临床表现均好转,体质量较模型组稍有增加,胃黏膜组织病理改变明显好转;灭幽汤低剂量组小鼠灌胃后症状、病理改变较之模型组没有显著变化。模型组胃黏膜细胞大量凋亡,除灭幽汤低剂量组外,其余药物干预组细胞凋亡情况较模型组显著减少。模型组小鼠胃组织中PTEN、FoxO3a蛋白及mRNA表达水平均明显高于对照组(P均<0.01),PI3K、p-Akt蛋白及PI3K、Akt mRNA表达水平均明显低于对照组(P均<0.01);胃四联组、灭幽汤中剂量组、灭幽汤高剂量组小鼠胃组织中PTEN、FoxO3a蛋白及mRNA表达水平均明显低于模型组(P均<0.01),PI3K、p-Akt蛋白及PI3K、Akt mRNA表达水平均明显高于�Objective It is to observe the effect of Mieyou Decoction(MD)on the expression of PTEN,PI3K,p-Akt,Akt,FoxO3a protein and mRNA in the gastric tissue of H.Pylori-associated gastritis(HAG)mice with spleen-stomach damp-heat syndrome,and to explore the possible mechanism of Mieyou Decoction in treating spleen-stomach damp-heat syndrome.Methods Sixty BALB/c mice were randomly divided into control group,model group,high-dose MD group,medium-dose MD group,low-dose MD group and stomach quadruple group.Except for the control group,the other groups were modeled by the compound etiology method.After successful modeling,the mice in the control group and model group were given distilled water by gavage,once per day;the mice in the low,medium and high dose groups of MD were respectively given MD 6.2 g/kg,12.4 g/kg,24.8 g/kg by gavage,once per day;the mice in the gastric quadruple group were given pantoprazole enteric-coated capsules+bismuth potassium citrate tablets+tinidazole tablets+clarithromycin tablets 280.6 mg/kg by gavage,once in the morning and evening.Each group was treated with intragastric administration for 14 days,and the general condition of mice in each group was observed.After the gavage,HE staining was used to observe the pathological manifestations of gastric mucosal tissue,TUNEL method was used to detect the apoptosis of gastric mucosal cells,Western blot was used to detect the expression of PTEN,PI3K,p-Akt,and FoxO3a in the gastric tissue,and RT-PCR method was used to detect the expression of PTEN,PI3K,Akt,FoxO3a mRNA in gastric tissue.Results The mice in the model group showed laziness,decreased appetite,loose and dull fur,reduced body mass,increased rectal temperature,and HE staining of gastric mucosa showed chronic superficial gastritis;these clinical manifestation were improved after given corresponding medicine by gavage in stomach quadruple group,medium and high dose of MD groups,their body weight increased slightly compared with the model group,and the pathological changes of gastric mucosa were signi

关 键 词：幽门螺杆菌 胃炎 灭幽汤 脾胃湿热证 细胞凋亡 PTEN-PI3K-Akt-FoxO信号通路 

分 类 号：R-332[医药卫生]