莪连颗粒对胃癌大鼠胃组织自噬及PI3K/Akt/mTOR信号通路的影响  

Effect of Elian Granule on Autophagy and PI3K/Akt/mTOR Signaling Pathway in Gastric Tissue of Rats with Gastric Cancer

作　　者：易志荣 林倚莉 王煜姣 贾庆玲 张钰琴[1] 谢天一[1] 凌江红[1,2] YI Zhi-rong;LIN Yi-li;WANG Yu-jiao;JIA Qing-ling;ZHANG Yu-qin;XIE Tian-yi;LING Jiang-hong(The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200021,China)

机构地区：[1]广西医科大学第一附属医院,南宁530021 [2]上海中医药大学附属曙光医院,上海200021

出　　处：《中国实验方剂学杂志》2021年第22期84-91,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81560763);2019国家中医药管理局《中医药循证能力建设项目》(2019XZZX-XH013)。

摘　　要：目的:探讨莪连颗粒对胃癌大鼠胃组织自噬及磷酯酰肌醇-3激酶(PI3K)/蛋白激酶B(PKB/Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的影响。方法:SPF级SD大鼠随机分为正常组、模型组、莪连颗粒组和胃复春组。除正常组常规饲养外,模型组、莪连颗粒组及胃复春组使用N-甲基-N’-硝基-N-亚硝基胍(MNNG)综合法诱导构建胃癌大鼠模型,并分别予以生理盐水、莪连颗粒水溶液(3.240 g·kg(-1))及胃复春水溶液(0.390 g·kg(-1))灌胃,连续48周。剖腹取胃,肉眼观察胃大体改变,采用苏木素-伊红(HE)染色观察大鼠胃组织病理学变化,实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测大鼠胃组织微管相关蛋白1轻链3β(LC3B),Beclin1,p62,PI3K,Akt,mTOR mRNA及蛋白表达。结果:与正常组比较,模型组大鼠胃胀大,胃壁变薄,胃黏膜色泽苍白,皱襞萎缩浅平,走行紊乱,可见结节及赘生物;与模型组比较,莪连颗粒组大鼠胃胀大减轻,胃壁变薄改善,胃黏膜色暗,皱襞减少,走行尚规整,偶见细颗粒状结节。HE染色显示,与正常组比较,模型组大鼠胃组织腺体排列拥挤紊乱,形态多样,细胞形态不一,胞浆嗜碱性,核大深染不规则,可见核分裂像,黏膜肌层浸润破坏;与模型组比较,莪连颗粒组大鼠胃组织腺体排列尚规整,少部分可见轻度异型细胞。与正常组比较,模型组大鼠胃组织LC3B,Beclin1 mRNA和蛋白表达明显降低(P<0.05),PI3K,p62,Akt及mTOR mRNA和蛋白表达明显升高(P<0.05);与模型组比较,莪连颗粒组胃组织LC3B,Beclin1 mRNA和蛋白表达明显升高(P<0.05),PI3K mRNA及p62,Akt,mTOR mRNA和蛋白表达明显降低(P<0.05)。结论:莪连颗粒可改善胃癌大鼠胃组织细胞异型性,其机制可能与抑制PI3K/Akt/mTOR信号通路促进自噬相关。Objective: To investigate the effect of Elian granule on autophagy and the phosphatidylinositol-3 kinase(PI3 K)/protein kinase B(PKB/Akt)/mammalian target of rapamycin(mTOR)signaling pathway in gastric tissue of rats with gastric cancer. Method: SPF SD rats were randomly divided into the normal,model,Elian granule,and Weifuchun groups. In addition to the routine feeding in the normal group, the model, Elian granule, and Weifuchun groups received N-methyl-N’-nitro-N-nitrosoguanidine(MNNG)to induce gastric cancer in rats,and they were respectively given normal saline,Elian granule aqueous solution(3.240 g·kg(-1))and Weifuchun aqueous solution(0.390 g·kg(-1))by gavage(ig)for 48 weeks.The gross changes of the stomach taken by laparotomy were observed by naked eyes. Hematoxylin-eosin(HE)staining was performed to observe the histopathological changes of the gastric tissue in rats. Real-time quantitative polymerase chain reaction(Real-time PCR)and Western blot(WB)were used to detect the mRNA and protein expression of microtubule-associated protein 1 light chain 3 beta(LC3 B),Beclin1,p62,PI3 K,Akt,mTOR in rat gastric tissue. Result: Compared with the normal group,the model group showed gastric distension,thinner gastric wall,pale gastric mucosa,atrophied and flat folds,disordered course,and visible nodules and vegetations. Compared with the model group,the Elian granule group demonstrated alleviated gastric distension,dark gastric mucosa,reduced folds,and regular course,with the thinned gastric wall improved and granular nodules observed occasionally. According to HE staining,compared with the normal group,the model group showed crowded and disordered rat gastric glands,diverse in shape,varied cell morphology,basophilic cytoplasm,large irregular hyperchromatic nuclei,visible mitosis,and infiltrated and destroyed muscularis mucosae. While compared with the model group,the arrangement of gastric glands was regular,and a few mildly atypical cells could be observed in rats of the Elian granule group. Compared with the no

关 键 词：莪连颗粒 胃癌 自噬 磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]