Lin Xian-Gang, Li Wei, Xiang Shui-Ying, Wu Sheng-Bing, Zhang Xin-Fang, Jiang Chuan-Wei, Liu Zi-Bing, Chen Ye-Nong
College of Acupuncture-moxibustion and Massage, Anhui University of Chinese Medicine, Hefei 230038, China.
Xin'an Medicine of the Ministry of Education of the People's Republic of China, Anhui University of Chinese Medicine, Hefei 230038, China.
Zhen Ci Yan Jiu. 2021 Mar 25;46(3):180-6. doi: 10.13702/j.1000-0607.200420.
To observe the effect of electroacupuncture (EA) at "Zusanli"(ST36) on the expression of epidermal growth factor receptor (EGFR), tumor necrosis factor α(TNF-α) transfer growth factor α(TGF-α), interleukin-8(IL-8), p38 mitogen-activated protein kinases (p38MAPK), mucin-5AC (MUC5AC) and other related factors in chronic obstructive pulmonary disease (COPD) rats, so as to reveal its underlying mechanisms in improving COPD.
A total of thirty male SD rats were randomly divided into normal control, model and EA groups, with 10 rats in each group. The COPD model was replicated using a combined method of tracheal infusion of lipopolysaccharide (LPS) and forced smoke-inhaling. EA (1-3 mA, 4 Hz/20 Hz) was applied to bilateral ST36 for 30 min, once daily for two consecutive weeks. The lung ventilation activities including the forced vital capacity (FVC) and forced expiratory volume (FEV) at 0.1 and 0.3 s (FEV0.1, FEV0.3) were detected. Histopathological changes of the middle lobe and bronchus of the right lung were observed after H.E. staining. The contents of TGF-α, TNF-α and IL-8 in the serum, bronchoalveolar lavage fluid (BALF) and superior lobe of the right lung were assayed by using ELISA, and the expression levels of EGFR, p38MAPK and MUC5AC proteins (inferior lobe of the left lung) and mRNAs (inferior lobe of the right lung) detected using Western blot, immunohistochemistry (strept avidin-biotin complex, SABC method) and real-time quantitative PCR, respectively.
Compared with the normal group, the FVC, FEV0.1, FEV0.3, FEV0.1/FVC and FEV0.3/FVC levels were significantly decreased (<0.01), while the contents of TNF-α, TGF-α and IL-8 in the serum, BALF and lung tissues, expression levels of EGFR, p38MAPK and MUC5AC mRNAs and proteins, and the immunoactivity of EGFR, p38MAPK and MUC5AC in the lung tissues were significantly increased in the model group (<0.01). After EA intervention, the decreased levels of the FVC, FEV0.1, FEV0.3, FEV0.1/FVC and FEV0.3/FVC, and the increased levels of the abovementioned genes and proteins were all reversed in the EA group (<0.01, <0.05). After modeling, the bronchial walls were thickened, with enlarged alveolar cavities, fractured alveolar walls, obvious inflammatory cell infiltration, and rich mucus secretion in the lumen, which was relatively milder in the EA group.
EA of ST36 can improve the ventilation function in COPD rats, which may be associated with its function in down-regulating the levels of TNF-α, TGF-α, IL-8, EGFR, p38MAPK and MUC5AC mRNAs and proteins in the lung tissues, inhibiting EGFR-p38MAPK signaling mediated expression of MUC5AC.
观察电针“足三里”（ST36）对慢性阻塞性肺疾病（COPD）大鼠表皮生长因子受体（EGFR）、肿瘤坏死因子α（TNF-α）、转化生长因子α（TGF-α）、白细胞介素-8（IL-8）、p38丝裂原活化蛋白激酶（p38MAPK）、黏蛋白5AC（MUC5AC）等相关因子表达的影响，以揭示其改善COPD的潜在机制。
将30只雄性SD大鼠随机分为正常对照组、模型组和电针组，每组10只。采用气管内注入脂多糖（LPS）并强迫烟熏的联合方法复制COPD模型。将电针（1 - 3 mA，4 Hz/20 Hz）施加于双侧ST36，持续30分钟，每日1次，连续两周。检测肺通气活动，包括用力肺活量（FVC）以及0.1秒和0.3秒时的用力呼气量（FEV）（FEV0.1、FEV0.3）。苏木精-伊红（H.E.）染色后观察右肺中叶和支气管的组织病理学变化。采用酶联免疫吸附测定（ELISA）法检测血清、支气管肺泡灌洗液（BALF）和右肺上叶中TGF-α、TNF-α和IL-8的含量，分别采用蛋白质印迹法、免疫组织化学法（链霉亲和素-生物素复合物，SABC法）和实时定量聚合酶链反应（PCR）检测EGFR、p38MAPK和MUC5AC蛋白（左肺下叶）和mRNA（右肺下叶）的表达水平。
与正常组相比，模型组FVC、FEV0.1、FEV0.3、FEV0.1/FVC和FEV0.3/FVC水平显著降低（<0.01），而血清、BALF和肺组织中TNF-α、TGF-α和IL-8的含量，肺组织中EGFR、p38MAPK和MUC5AC mRNA及蛋白的表达水平，以及EGFR、p38MAPK和MUC5AC的免疫活性均显著升高（<0.01）。电针干预后，电针组FVC、FEV0.1、FEV0.3、FEV0.1/FVC和FEV0.3/FVC降低的水平以及上述基因和蛋白升高的水平均得到逆转（<0.01，<0.05）。造模后，支气管壁增厚，肺泡腔扩大，肺泡壁断裂，炎症细胞浸润明显，管腔内黏液分泌丰富，电针组情况相对较轻。
电针ST36可改善COPD大鼠的通气功能，这可能与其下调肺组织中TNF-α、TGF-α、IL-8、EGFR、p38MAPK和MUC5AC mRNA及蛋白水平，抑制EGFR - p38MAPK信号介导的MUC5AC表达有关。
