针刺“大椎-身柱”对穴通过DEK激活PI3K/AKT通路干预过敏性哮喘大鼠上皮间质转化的研究  

Acupuncture inhibits epithelial mesenchymal transition in allergic asthma rats by activating the PI3K/AKT pathway through DEK activation

作　　者：苗镡允 吕玉娥 吉美奇 尹浩 吴涵 胡志祥 翟春涛 李蕾 MIAO Xin-yun;LÜYu-e;JI Mei-qi;YIN Hao;WU Han;HU Zhi-xiang;ZHAI Chun-tao;LI Lei(Shanxi University of Chinese Medicine,Jinzhong 030600,Shanxi province,China;Shanxi Acupuncture Hospital,Taiyuan 030006)

机构地区：[1]山西中医药大学,山西晋中030600 [2]山西省针灸医院,太原030006

出　　处：《针刺研究》2025年第2期174-182,共9页Acupuncture Research

基　　金：山西省“四个一批”科技兴医创新计划重点攻关专项项目(No.2022XM35);山西中医药大学创新团队项目(No.2022TD1006);山西省科技创新人才团队项目(No.200204051002011);吕景山国医大师“对法”传承省技术创新中心项目(No.2022D100089052823242891545041)。

摘　　要：目的:观察针刺对穴“大椎-身柱”对过敏性哮喘(AA)大鼠DEK/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)通路及上皮间质转化相关因子表达的影响,探讨针刺治疗AA的潜在机制。方法:60只SD大鼠随机分为空白组、模型组、对穴组、地塞米松组、假针组,每组12只。通过腹腔注射及雾化吸入卵清蛋白(OVA)制备AA模型。对穴组大鼠予针刺对穴“大椎-身柱”,留针20 min,每日1次,连续14 d;地塞米松组大鼠腹腔注射0.5 mg/kg地塞米松溶液,每日1次,连续14 d;假针组针刺非经非穴点,操作同对穴组,连续14 d。干预结束后,HE染色法观察大鼠肺组织病理形态改变;ELISA法检测大鼠肺泡灌洗液(BALF)中白细胞介素(IL)-4、IL-5及血清免疫球蛋白IgE、IgM水平;Western blot法检测大鼠肺组织中DEK、磷酸化(p)-PI3K、p-AKT、E钙黏蛋白(E-cadherin)蛋白相对表达量;免疫荧光染色法检测大鼠肺组织中DEK、p-PI3K、p-AKT、E-cadherin阳性表达。结果:与空白组比较,模型组和假针组大鼠肺组织支气管壁增厚,炎性细胞浸润增多;对穴组及地塞米松组支气管壁增厚减少,炎性细胞浸润程度降低。与空白组比较,模型组BALF中IL-4、IL-5及血清IgE、IgM含量显著升高(P<0.01),肺组织中DEK、p-PI3K、p-AKT蛋白表达量升高(P<0.01),DEK、p-PI3K和p-AKT阳性表达水平升高(P<0.01),E-cadherin蛋白表达量和阳性表达水平均降低(P<0.01)。与模型组比较,对穴组及地塞米松组BALF中IL-4、IL-5及血清IgE、IgM含量均降低(P<0.01,P<0.05),肺组织DEK、p-PI3K、p-AKT蛋白表达量降低(P<0.01),E-cadherin蛋白表达量升高(P<0.01),肺组织DEK、p-PI3K阳性表达水平降低(P<0.05,P<0.01),E-cadherin阳性表达水平升高(P<0.05);对穴组p-AKT阳性表达水平降低(P<0.05);假针组BALF中IL-5含量降低(P<0.01)。结论:针刺对穴“大椎-身柱”可以改善OVA诱导的AA大鼠气道重塑改变,其作用机制可能是通过下调DEK表达,抑制PI3K/AKT激活,延缓Objective To observe the effect of acupuncture at“Dazhui”(GV14)and“Shenzhu”(GV12)on DEK/phosphorylated phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway and epithelial-mesenchymal transition related factors in rats with allergic asthma(AA),so as to explore its potential mechanism in the treatment of AA.Methods Sixty SD rats were randomly divided into normal,model,acupuncture,dexamethasone(DEX)and sham-acupuncture groups,with 12 rats in each group.The AA model was made by intraperitoneal injection of mixture solution of ovalbumin and aluminium hydroxide gel(on day 1 and 7)and followed by inhalation of atomized 1%ovalbumin(30 min,from day 15,once daily for 14 days).Rats in the acupuncture group received acupuncture at GV14 and GV12,while rats in the sham-acupuncture group received acupuncture at the non-meridian and non-acupoint point,both for 20 min,once daily for 14 days.Rats of the DEX group received intraperitoneal injection of DEX(0.5 mg/kg),once daily for 14 days.H.E.staining was used to observe the changes of pathological morphology in lung tissue.ELISA was used to detect the levels of interleukin(IL)-4 and IL-5 in broncho alveolar lavage fluid(BALF),and serum immunoglobulin IgE and IgM.Western blot was used to detect the expressions of DEK,PI3K,AKT,phosphorylated(p)-PI3K,p-AKT,and E-cadherin in lung tissue.Immunofluorescence staining was used to detect the positive expressions of DEK,p-PI3K,p-AKT,and E-cadherin in lung tissue.Results Compared with the normal group,the levels of IL-4,IL-5 in BALF and serum IgE and IgM,the protein expression levels of DEK,p-PI3K and p-AKT in lung tissue,the positive expression levels of DEK,p-PI3K and p-AKT were increased in the model group(P<0.01),while the protein expression and positive expression levels of E-cadherin were decreased(P<0.01).In comparison with the model group,the contents of IL-4,IL-5 in BALF and serum IgE and IgM,the protein expressions of DEK,p-PI3K and p-AKT,the positive expression levels of DEK and p-PI3K in the acupuncture and DE

关 键 词：针刺 过敏性哮喘 上皮间质转化 DEK 磷酸化磷脂酰肌醇3-激酶 蛋白激酶B 

分 类 号：R245[医药卫生—针灸推拿学]