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[电针“夹脊”(EX-B2)通过减轻气道炎症和调节过敏性哮喘大鼠的PI3K/AKT/mTOR信号通路来改善哮喘]

[Electroacupuncture of "Jiaji"(EX-B2) improves asthma by reducing airway inflammation and regulating PI3K/AKT/mTOR signaling pathway in rats with allergic asthma].

作者信息

Zhu Tao, Li Jie, Wan Hong-Ye, Chang Bu, Cheng Yan-Ting, Ji Yu-Fang, Ji Lai-Xi

机构信息

College of Acupuncture-Moxibustion and Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.

The Second Clinical Medicine College, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province.

出版信息

Zhen Ci Yan Jiu. 2025 Jun 25;50(6):666-674. doi: 10.13702/j.1000-0607.20241336.

Abstract

OBJECTIVES

To observe the effect of electroacupuncture (EA) at "Jiaji" (EX-B2) on asthma symptoms, airway inflammation and phosphatidylinositol 3-kinase (PI3K) / protein kinase B (AKT) / mammalian target of rapamycin (mTOR) signaling in rats with allergic asthma (AA), so as to explore its mechanisms underlying improvement of AA.

METHODS

Forty male SD rats were randomly divided into blank control, AA model, EA and medication groups, with 10 rats in each group. The AA model was established by intraperitoneal injection of 1 mL 1% ovalbumin (OVA, grade V) suspension on the 1 day and 7 day and inhalation of 1% aerosol OVA (on the 15 day) for 30 min, once daily for 2 weeks. Rats of the EA group received EA stimulation (2 Hz/15 Hz, 1-2 mA) of bilateral EX-B2 for 20 min, once daily for 2 weeks. Rats of the medication group received intraperitoneal injection of dexamethasone solution (0.5 mg/kg), once daily for 2 weeks. The nasal scratching or tickling (0-3 points) and asthma (0-9 points) behavior were graded, and their summation (behavior score) was used for assessing the severity of asthma. Histopathological changes of the lung tissue were assessed by giving pathological score after H.E. staining. The contents of IL-4, IL-5, IL-13 in the bronchoalveolar lavage fluid (BALF) and lgE in the serum were detected by ELISA. The immunoactivity of Beclin1 and microtubule-associated protein 1A/1B light chain 3B (LC3B) in the lung tissue was detected by immunohistochemistry. The ratio of expression levels of p-PI3K/PI3K, p-AKT/AKT, p-mTOR /mTOR proteins in the lung tissue were detected by Western blot.

RESULTS

Compared with the blank control group, the model group showed an increase in the behavior score (shortness of breath, wheezing and irritability, etc), pathological scores of the lung tissue, contents of IL-4, IL-5 and IL-13 in the BALF and IgE in the serum, the immunoactivity of Beclin1 and LC3B in the lung tissue (<0.01), and a decrease in the body weight, and the ratio of expression levels of p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR proteins in the lung tissue (<0.01). In comparison with the model group, the behavior score, pathological score, the contents of IL-4, IL-5, IL-13 and IgE, and the immunoactivity of Beclin1 and LC3B were significantly decreased (<0.05, <0.01), while the ratio of expression levels of p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR proteins were considerably increased (<0.05, <0.01) in both EA and medication groups. The body weight of rats was increased (<0.05) in the EA group compared to the model group. No significant differences were found between the EA and medication groups of all the indexes mentioned above.

CONCLUSIONS

EA of EX-B2 can improve the symptoms of asthma in rats with AA, which may be associated with its functions in inhibiting the autophagy level of airway epithelial cells in the lung tissue and reducing airway inflammation by regulating PI3K/AKT/mTOR signaling pathway.

摘要

目的

观察电针“夹脊”(EX - B2)对过敏性哮喘(AA)大鼠哮喘症状、气道炎症及磷脂酰肌醇3 -激酶(PI3K)/蛋白激酶B(AKT)/雷帕霉素靶蛋白(mTOR)信号通路的影响,以探讨其改善AA的作用机制。

方法

40只雄性SD大鼠随机分为空白对照组、AA模型组、电针组和药物组,每组10只。于第1天和第7天腹腔注射1 mL 1%卵清蛋白(OVA,Ⅴ级)混悬液,第15天吸入1% OVA气雾剂30分钟,每日1次,连续2周,建立AA模型。电针组大鼠给予双侧EX - B2电针刺激(2 Hz/15 Hz,1 - 2 mA)20分钟,每日1次,连续2周。药物组大鼠腹腔注射地塞米松溶液(0.5 mg/kg),每日1次,连续2周。对鼻搔抓或瘙痒(0 - 3分)和哮喘(0 - 9分)行为进行评分,二者之和(行为评分)用于评估哮喘严重程度。苏木精 - 伊红(H.E.)染色后给予病理评分,评估肺组织的组织病理学变化。采用酶联免疫吸附测定(ELISA)法检测支气管肺泡灌洗液(BALF)中白细胞介素 - 4(IL - 4)、白细胞介素 - 5(IL - 5)、白细胞介素 - 13及血清中免疫球蛋白E(IgE)的含量。采用免疫组织化学法检测肺组织中自噬相关蛋白Beclin1和微管相关蛋白1A/1B轻链3B(LC3B)的免疫活性。采用蛋白质免疫印迹法检测肺组织中p - PI3K/PI3K、p - AKT/AKT、p - mTOR/mTOR蛋白表达水平的比值。

结果

与空白对照组比较,模型组行为评分(气短、喘息、烦躁等)、肺组织病理评分、BALF中IL - 4、IL - 5、IL - 13含量及血清中IgE含量、肺组织中Beclin1和LC3B免疫活性升高(P < 0.01),体重下降,肺组织中p - PI3K/PI3K、p - AKT/AKT、p - mTOR/mTOR蛋白表达水平的比值降低(P < 0.01)。与模型组比较,电针组和药物组行为评分、病理评分、IL - 4、IL - 5、IL - 13及IgE含量、Beclin1和LC3B免疫活性均显著降低(P < 0.05,P < 0.01),肺组织中p - PI3K/PI3K、p - AKT/AKT、p - mTOR/mTOR蛋白表达水平的比值显著升高(P < 0.05,P < 0.01)。电针组大鼠体重较模型组增加(P < 0.05)。上述各指标电针组与药物组比较,差异均无统计学意义。

结论

电针夹脊穴可改善AA大鼠的哮喘症状,其机制可能与抑制肺组织气道上皮细胞自噬水平及通过调节PI3K/AKT/mTOR信号通路减轻气道炎症有关。

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