基于PI3K/Akt通路探讨芪灵方对慢性萎缩性胃炎伴肠化大鼠的保护作用  

Protective Effect of Qiling Prescription on Rats with Chronic Atrophic Gastritis Combined with Gastric Intestinal Metaplasia Via PI3K/Akt Pathway

作　　者：黄项鸣 樊欣钰 陆敏[1,2,3] HUANG Xiangming;FAN Xinyu;LU Min(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,China;Lishui District Hospital of Traditional Chinese Medicine,Nanjing 211299,China;Clinical College,Jiangsu Vocational College of Medicine,Nanjing 210029,China)

机构地区：[1]南京中医药大学附属中西医结合医院,南京210028 [2]南京市溧水区中医院,南京211299 [3]江苏卫生健康职业学院临床学院,南京210029

出　　处：《中国实验方剂学杂志》2024年第22期79-86,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江苏省中医药管理局中医消化肿瘤临床医学创新中心项目(苏中医科教[2021]6号);江苏省研究生科研与实践创新计划项目(KYCX232182);江苏卫生健康职业学院“医教协同、中西并举”专项(YJXTL202309)。

摘　　要：目的:探讨芪灵方干预慢性萎缩性伴肠上皮化生(GIM)的潜在作用机制。方法:将80只SPF级SD大鼠随机分为8组(每组10只):空白组、空白加芪灵方组、模型组、芪灵方高、中、低剂量组、叶酸组、摩罗丹组,除空白组、空白加芪灵方组外,其余各组大鼠使用0.02 mol·L-1的N-甲基-N′-硝基-N-亚硝基胍(MNNG)溶液灌胃,并佐以饥饱失常法造模。造模成功后,空白组、模型组予蒸馏水,空白加芪灵方组、芪灵方高、中、低剂量组分别予7.60、15.21、7.60、3.80 g·kg-1芪灵方水煎剂,叶酸组予0.002 g·kg-1叶酸悬浊液,摩罗丹组予1.40 g·kg-1摩罗丹悬浊液灌胃,1次/d,给药8周,期间观察大鼠的一般情况与体质量。麻醉后取大鼠胃组织行苏木素-伊红(HE)染色;免疫组化法(IHC)检测胃组织黏蛋白2(MUC2)、尾部型同源框转录因子2(CDX2)水平;酶联免疫吸附测定法(ELISA)检测血清中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、胱天蛋白酶-3(Caspase-3)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)水平;蛋白免疫印迹法(Western blot)检测胃组织PI3K、Akt及磷酸化水平。结果:动物实验显示,与空白组比较,参与造模的大鼠体质量自16周起有下降趋势;与模型组比较,芪灵方高、中剂量对大鼠精神状态、体质量均有改善作用。病理结果显示,第24周造模成功,与空白组比较,参与造模的大鼠胃黏膜固有腺体减少,排列相对紊乱;与模型组比较,芪灵方高、中剂量组杯状细胞明显减少或消失,可改善大鼠胃黏膜病理。与空白组比较,模型组胃组织MUC2、CDX2水平升高(P<0.01);与模型组比较,芪灵方高、中剂量可明显降低胃组织MUC2、CDX2水平(P<0.05,P<0.01)。与空白组比较,模型组大鼠血清中IL-6、IL-1β、TNF-α、Caspase-3、PI3K、Akt水平均提高(P<0.05);与模型组比较,高剂量芪灵方可显著降低血清中IL-1β、Caspase-3、PI3K、Akt含量(P<0.01),中Objective:To explore the potential mechanism of Qiling prescription in intervening in chronic atrophic gastritis with gastric intestinal metaplasia(GIM).Method:The 80 SPF-grade SD rats were randomly divided into the following eight groups(10 rats per group):blank group,blank+Qiling prescription group,model group,high-dose Qiling prescription group,medium-dose Qiling prescription group,low-dose Qiling prescription group,folic acid group,and morodan group.Except for the blank and blank+Qiling prescription groups,the other groups underwent modeling by intragastric administration of 0.02 mol·L-1 N-methyl-N′-nitro-N-nitrosoguanidine(MNNG)solution combined with irregular feeding.After successful modeling,the blank and model groups were given distilled water,the blank+Qiling prescription group,and high,medium,and low-dose Qiling prescription groups were given Qiling prescription water decoction at 7.60,15.21,7.60,3.80 g·kg-1,respectively,the folic acid group was given folic acid suspension at 0.002 g·kg-1,the morodan group was given morodan suspension at 1.40 g·kg-1 by gavage once a day for 8 weeks.The general condition and body weight of the rats were observed during the experiment.Hematoxylin-eosin(HE)staining was performed on gastric tissues.Immunohistochemistry(IHC)was used to detect the levels of mucin 2(MUC2)and caudal-type homeobox transcription factor 2(CDX2)in gastric tissues.Enzyme-linked immunosorbent assay(ELISA)was used to measure the serum levels of interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),cysteine-aspartic protease-3(Caspase-3),phosphatidylinositol 3-kinase(PI3K),and protein kinase B(Akt).Western blot analysis was performed to detect the expression and phosphorylation levels of PI3K and Akt in gastric tissues.Result:Animal experiments showed that compared to the blank group,the rats in the model group had a trend of weight loss starting from week 16.Compared to the model group,high and medium doses of Qiling prescription improved the mental state and body weight o

关 键 词：慢性萎缩性胃炎伴肠上皮化生 磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)通路 炎癌转化 芪灵方 黄芪建中汤 紫芝丸 胃癌前病变 

分 类 号：R2-0[医药卫生—中医学] R33R289R318.14