隔药饼灸对慢性萎缩性胃炎大鼠PTEN-AKT抑癌途径的作用机制研究  

Mechanism Study of the Effect of Herb-partitioned Moxibustion on PTEN-AKT Tumor Suppressor Pathway in Rats with Chronic Atrophic Gastritis

作　　者：郑雪 萧有智 吴梦蝶 吴焕淦[3,4] 马喆[3,4] 王文佳 黄艳[3,4] 李琪[3,4] 李璟 ZHENG Xue;XIAO You-zhi;WU Meng-die;WU Huan-gan;MA Zhe;WANG Wen-jia;HUANG Yan;LI Qi;LI Jing(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China)

机构地区：[1]上海中医药大学,上海201203 [2]河南中医药大学第一附属医院,郑州450000 [3]上海中医药大学附属岳阳中西医结合医院,上海200437 [4]上海市针灸经络研究所,上海200030

出　　处：《上海针灸杂志》2021年第12期1500-1508,共9页Shanghai Journal of Acupuncture and Moxibustion

基　　金：国家自然科学基金项目(81674071);上海市临床重点专科项目(shslczdzk04701);国家中医药管理局针灸治疗优势病的循证研究(2019XZZX-ZJ0011);上海市针灸临床医学研究中心(20MC1920500)。

摘　　要：目的探索艾灸通过调控PTEN-AKT抑癌途径相关基因的表达,以达到修复慢性萎缩性胃炎(CAG)受损胃黏膜的可能机制。方法将52只雄性Wistar大鼠随机分为正常组和造模组,造模组采用N-甲基-N-硝基-N-亚硝基胍(MNNG)诱导剂结合夹尾刺激与饥饱失常法制备CAG大鼠模型,34周模型鉴定成功后,将造模大鼠随机分为模型组、隔药饼灸组和西药组,每组10只。隔药饼灸组取中脘和气海穴,每日1次,每周6次;西药组予叶酸悬浊液灌胃,每日1次,每周6次。干预4周后取材,采用HE染色法观察胃窦组织病理学变化,免疫组化法检测各组大鼠胃窦组织PTEN、p-AKT、Caspase-9、PIP2、MDM2的蛋白表达,RT-qPCR法检测各组大鼠胃窦组织PTEN、PI3KCA、Caspase-9、p53 mRNA的表达。结果与正常组比较,模型组可见固有腺体萎缩、肠上皮化生和异型增生;与模型组比较,隔药饼灸组和西药组固有腺体排列较规整,固有腺体萎缩、肠上皮化生、异型增生程度减轻。与正常组比较,模型组大鼠胃窦组织PTEN、Caspase-9蛋白和mRNA表达均显著降低(P<0.05),p-AKT、PIP2、MDM2蛋白表达均显著升高(P<0.05);与模型组比较,西药组和隔药饼灸组胃窦组织PTEN、Caspase-9蛋白表达和Caspase-9、PI3KCA mRNA表达均显著升高(P<0.05),p-AKT、PIP2、MDM2蛋白表达均显著降低(P<0.05),西药组胃窦组织PTEN的mRNA表达升高(P<0.05)。结论隔药饼灸中脘和气海穴可改善CAG大鼠的胃窦组织形态学变化,修复胃黏膜的损伤;隔药饼灸可能通过调控CAG大鼠胃窦组织PTEN-AKT抑癌途径有效控制CAG的胃黏膜损伤,是隔药饼灸治疗CAG和防治癌前病变的效应机制之一。Objective To explore the plausible mechanism of moxibustion in repairing the damaged gastric mucosa in chronic atrophic gastritis(CAG)via targeting the expression of genes related to PTEN-AKT tumor suppressor pathway.Method Fifty-two male Wistar rats were randomly divided into a normal group and a modeling group.Rats in the modeling group were established to be CAG models using N-methyl-N-nitro-N-nitrosoguanidine(MNNG)inducer combined with tail-clamping stimulation and irregular feeding method.At the 34th week,when the CAG model was confirmed a success,the model rats were randomly divided into a model group,an herb-partitioned moxibustion group,and a Western medication group,with 10 rats in each group.The herb-partitioned moxibustion group received treatment at Zhongwan(CV12)and Qihai(CV6),once a day,six times a week.The Western medication group was given folic acid suspension by gavage,once a day,six times a week.The samples were collected after four-week intervention.The histopathological changes of gastric antrum were observed using hematoxylin-eosin(HE)staining;the protein expression levels of PTEN,p-AKT,Caspase-9,PIP2,and MDM2 in gastric antrum tissue in each group were detected by the immunohistochemical method;the mRNA expression levels of PTEN,PI3KCA,Caspase-9,and p53 in gastric antrum tissue were examined by RT-qPCR method.Result Compared with the normal group,atrophic proper glands and intestinal metaplasia and dysplasia were found in the model group;compared with the model group,the proper glands were arranged more regularly,and the atrophy of proper glands and intestinal metaplasia and dysplasia were alleviated in the herb-partitioned moxibustion group and the Western medication group.Compared with the normal group,the protein and mRNA expression levels of PTEN and Caspase-9 in rat’s gastric antrum tissue were significantly reduced(P<0.05),and the protein expression levels of p-AkT,PIP2,and MDM2 were significantly increased(P<0.05)in the model group.Compared with the model group,the protein expressi

关 键 词：针灸疗法 灸法 药饼灸疗法 胃炎 萎缩性 磷酸酶与张力蛋白同源物 抑癌途径 大鼠 

分 类 号：R2-03[医药卫生—中医学]