基于PI3K/AKT/mTOR信号通路对电针干预慢性结肠炎大鼠的腧穴特异性及机制研究  

A study of acupoint specificity and mechanism of electroacupuncture intervention on chronic colitis in rats based on PI3K/AKT/mTOR signaling pathway

作　　者：唐坤鹏 吕嘉琪 文坛 张春青 马梦娜 任华山 闫丽萍 TANG Kun-peng;LV Jia-qi;WEN Tan;ZHANG Chun-qing;MA Meng-na;REN Hua-shan;YAN Li-ping(Shanxi University of Chinese Medicine,J inzhong 030619,China)

机构地区：[1]山西中医药大学,山西晋中030619

出　　处：《海南医学院学报》2023年第9期674-680,共7页Journal of Hainan Medical University

基　　金：山西省应用基础研究项目(201801D121358)。

摘　　要：目的:基于PI3K/AKT/mTOR信号通路探讨电针“天枢”、“大肠俞”、“足三里”、“上巨虚”穴对慢性实验性结肠炎大鼠的效应差异及作用机制。方法:将60只清洁级SD大鼠随机分为正常组、模型组、天枢组、大肠俞组、足三里组、上巨虚组,每组各10只,采用免疫致敏法结合局部刺激制备慢性结肠炎大鼠模型;各腧穴组于造模结束后行电针干预,疏密波,频率2 Hz/50 Hz,电流强度2 mA,每日1次,20 min/d。分别观察各组大鼠结肠组织黏膜炎症损伤情况;ELSA检测大鼠血清中促炎因子IL-23、IL-17、IL-10水平;RT-PCR检测结肠组织中PI3K、Akt、mTOR mRNA的表达;Western blot检测结肠组织中p-PI3k/PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白含量。结果:与正常组相比,模型大鼠一般状态较差,结肠黏膜受损严重,有大量炎性细胞浸润;血清中IL-23、IL-17表达水平明显升高,IL-10表达水平明显降低(P<0.01);结肠组织中PI3K、Akt、mTOR mRNA及p-PI3K、p-Akt、p-mTOR蛋白表达水平显著升高(P<0.05,P<0.01);与模型组相比,各腧穴电针干预组大鼠病理切片显示,结肠黏膜炎症反应及组织损伤均明显改善,血清中IL-23、IL-17含量明显降低(P<0.01),血清中IL-10表达水平明显上升(P<0.01),结肠组织中PI3K、Akt、mTOR mRNA及p-PI3K、p-Akt、p-mTOR蛋白表达明显降低(P<0.05,P<0.01);各腧穴组间相比,上巨虚组结肠黏膜各层恢复程度更接近正常组,疗效相对最好,上巨虚组与天枢、大肠俞、足三里组比较,其血清中IL-23、IL-17水平明显偏低(P<0.05),IL-10表达水平明显偏高(P<0.01);结肠组织PI3K、Akt、mTOR mRNA及p-PI3K、p-Akt、p-mTOR蛋白表达明显降低(P<0.05,P<0.01)。结论:电针天枢、大肠俞、足三里、上巨虚穴均可缓解慢性结肠炎大鼠的结肠黏膜损伤及炎症反应;其中,上巨虚对慢性结肠炎治疗的总体效应优于天枢、大肠俞、足三里;其作用机制可能与抑制PI3K/Akt/mTOR信号通路相关。Objective:To elucidate the differences in effects and mechanisms of action of electric‑needle therapy at Tian Shu(ST25),Da Chang Shu(BL25),Zu San Li(ST36)and Shang Ju Xu(ST37)acupoints on chronic experimental colitis in rats through the PI3K/AKT/mTOR signaling pathway.Methods:Sixty pathogen‑free SD rats were randomly assigned to six groups:the normal,model,Tian Shu(ST25),Da Chang Shu(BL25),Zu San Li(ST36)and Shang Ju Xu(ST37)groups,each with 10 rats.Chronic colitis was induced in rats by combining immunization and local stimulation.After model establishment,electrical needle intervention combined with dispersing wave of 2 Hz/50 Hz with a current intensity of 2 mA once daily for 20 min was applied on acupoints of each group.Subsequently,the inflammation of colonic mucosa and serum levels of inflammatory factors(IL‑23,IL‑17,IL‑10)were observed;ELISA was used to detect mRNA expressions of PI3K,Akt and mTOR in colitic tissues by RT‑PCR as well as protein content of p‑PI3k/PI3K,p‑Akt/Akt,and p‑mTOR/mTOR in colitic tissues by Western blotting.Results:Compared with the normal group,the model rats showed a poor general condition,serious damage to the colonic mucosa with a large number of inflammatory cells infiltration.The serum IL‑23 and IL‑17 expressions were significantly increased(P<0.01),while the serum IL‑10 expression was significantly decreased(P<0.01);the mRNA and protein expressions of PI3K,Akt,mTOR and p‑PI3K,p‑Akt and p‑mTOR were significantly increased(P<0.05,P<0.01).Compared with the model group,the pathological slices of rats in each acupoints intervention group showed obvious improvement of colitis inflammatory reaction and tissue damage;the serological levels of IL‑23 and IL‑17 were significantly reduced(P<0.01),while the serology level of IL‑10 was significantly increased(P<0.01);the expressions of PI3K,Akt,mTOR mRNA and p‑PI3K,p‑Akt,p‑mTOR proteins were significantly decreased(P<0.05,P<0.01).Compared with Tian Shu(ST25),Da Chang Shu(BL25),Zu San Li(ST36)groups,

关 键 词：电针 慢性结肠炎 腧穴特异性 PI3K/Akt/mTOR信号通路 

分 类 号：R245.97[医药卫生—针灸推拿学]