电针通过迷走神经调控肺泡巨噬细胞极化对慢性阻塞性肺疾病小鼠肺部炎症的影响

Electroacupuncture Modulates Alveolar Macrophage Polarization via the Vagus Nerve to Alleviate Pulmonary Inflammation in a Mouse Model of Chronic Obstructive Pulmonary Disease

  • 摘要:
    目的 探究电针对慢性阻塞性肺疾病(COPD)小鼠的抗炎效应及其潜在机制。
    方法 40只C57BL/6小鼠随机分为正常组、模型组、电针组、离断组、离断+电针组,每组8只。除正常组外,其余各组经香烟烟雾暴露12周建立小鼠COPD模型。造模后,离断组、离断+电针组在电针前行左侧颈部迷走神经切断术。电针组、离断+电针组进行电针治疗,选取肺俞和足三里穴,每日电针1次、每次20 min,共14 d。电针结束后,肺功能分析系统检测小鼠肺通气功能;HE染色观察肺组织病理;ELISA检测肺组织中白细胞介素(IL)-6、肿瘤坏死因子-α(TNF-α)、IL-10、转化生长因子-β(TGF-β)的含量;Western blot检测肺组织CD86、CD206蛋白的表达;流式细胞术检测肺组织F4/80+CD86+(M1型)和F4/80+CD206+(M2型)细胞的比例分布;免疫荧光技术观察CD86和CD206在肺组织中的表达。
    结果 与正常组比较,模型组小鼠肺功能各项指标明显下降(P < 0.01),肺病理呈明显损伤;M1占比,IL-6、TNF-α、CD86含量及表达显著升高(P < 0.05, P < 0.01),M2占比,IL-10、TGF-β、CD206含量及表达显著降低(P < 0.01)。与模型组比较,电针组肺功能及病理均有不同程度改善;M1占比,IL-6、TNF-α、CD86均有不同程度降低(P < 0.05, P < 0.01),M2占比,IL-10、TGF-β、CD206均有不同程度升高(P < 0.05, P < 0.01);离断组肺功能及病理均有降低或加重,IL-6、TNF-α、CD86含量及表达不同程度升高(P < 0.05, P < 0.01),IL-10、TGF-β、CD206含量及表达降低(P < 0.05, P < 0.01)。与电针组比较,离断+电针组M1占比,IL-6、TNF-α、CD86含量及表达升高(P < 0.01),M2占比,IL-10、TGF-β、CD206含量及表达降低(P < 0.05,P < 0.01)。
    结论 电针肺俞、足三里穴可以改善COPD小鼠肺功能和肺部炎症,促进肺泡巨噬细胞由M1向M2极化,其作用与迷走神经介导有关。

     

    Abstract:
    OBJECTIVE To investigate the anti-inflammatory effect of electroacupuncture in mice with chronic obstructive pulmonary disease (COPD) and its underlying mechanisms.
    METHODS Forty C57BL/6 mice were randomly divided into normal group, model group, electroacupuncture (EA) group, vagotomy group, and vagotomy+EA group, with 8 mice in each group. Except for the normal group, all groups were exposed to cigarette smoke for 12 weeks to establish a COPD model. After model establishment, the vagotomy group and the vagotomy+EA group underwent left cervical vagotomy before EA. EA treatment was performed at the Feishu (BL13) and Zusanli (ST36) acupoints once daily for 20 minutes for a total of 14 days. After EA, the pulmonary ventilation function of the mice was detected by a pulmonary function analysis system; lung tissue pathology was observed by HE staining; the levels of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), IL-10, and transforming growth factor-β (TGF-β) in lung tissue were detected by ELISA; the expression of CD86 and CD206 proteins in lung tissue was detected by Western blot; the distribution of F4/80+CD86+ (M1 type) and F4/80+CD206+ (M2 type) cells in lung tissue was determined by flow cytometry; the expression of CD86 and CD206 in lung tissue was observed by immunofluorescence.
    RESULTS Compared with the normal group, the model group showed significantly decreased lung function (P < 0.01), obvious lung pathological damage, increased M1 proportion, IL-6, TNF-α, CD86 content and expression (P < 0.05, P < 0.01), and decreased M2 proportion, IL-10, TGF-β, CD206 content and expression (P < 0.01). Compared with the model group, the EA group showed varying degrees of improvement in lung function and pathology; the M1 proportion, IL-6, TNF-α, and CD86 were reduced (P < 0.05, P < 0.01), while the M2 proportion, IL-10, TGF-β, and CD206 were increased (P < 0.05, P < 0.01). The vagotomy group showed worsened lung function and pathology, increased IL-6, TNF-α, and CD86 content and expression (P < 0.05, P < 0.01), and decreased IL-10, TGF-β, and CD206 content and expression (P < 0.05, P < 0.01). Compared with the EA group, the vagotomy+EA group showed increased M1 proportion, IL-6, TNF-α, and CD86 content and expression (P < 0.01), and decreased M2 proportion, IL-10, TGF-β, and CD206 content and expression (P < 0.05, P < 0.01).
    CONCLUSION EA at Feishu(BL13) and Zusanli(ST36) acupoints can improve lung function and pulmonary inflammation in COPD mice, promoting the polarization of alveolar macrophages from M1 to M2, which is mediated by the vagus nerve.

     

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