Qin Zhong-Yin, Chen Pan-Bi, Tang Xu-Yun, Du Di-Jia, Long Run-Jin
Graduate School, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China.
College of Acupuncture and Tuina, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China.
Zhen Ci Yan Jiu. 2022 Aug 25;47(8):690-5. doi: 10.13702/j.1000-0607.20210800.
To investigate the effects of acupuncture on phosphorylated P38 mitogen-activated protein kinase (p-P38MAPK), intercellular adhesion molecule-1 (ICAM-1), interferon γ (IFN-γ), and eosinophilic granulocytes (EOS) in lung tissue of asthmatic rats, and to explore the mechanism of acupuncture in regulating the apoptosis of EOS.
Clean-grade male Wistar rats were randomly divided into normal, model, dexamethasone and acupuncture groups, 8 rats in each group. The asthmatic model was established by intraperitoneal injection of mixture suspension (1 mL) of 10% ovalbumin and 10% Al(OH)_3+ normal saline, followed by inhalation of atomized 1% ovalbumin solution for 30 min, once daily for 2 weeks to trigger occurrence of asthmatic symptoms. The rats in dexamethasone group were intraperitoneally injected with 0.9 mg/kg dexamethasone since day 15 once a day for two consecutive weeks. In the acupuncture group, bilateral "Feishu" (BL13), "Pishu" (BL20), "Shenshu" (BL23), "Dingchuan" (EX-B1), and "Danzhong" (CV17) were selected for acupuncture treatment once every other day since day 15 for two consecutive weeks. Uniform reinforcing and reducing manipulation was carried out, and the needles were retained for 30 min. The pathological changes of the lung tissue were observed by hematoxylin-eosin (HE) staining. The apoptosis of EOS in the lung tissue of rats was detected by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. The expression of p-P38MAPK in the lung tissue was detected by Western blot. The protein and mRNA levels of ICAM-1 and IFN-γ in the lung tissue were determined by immunohistochemistry and real-time PCR, respectively.
The results of HE staining showed that the pulmonary alveoli and surrounding tissues were intact with no inflammatory cell infiltration in the normal group. The model group showed massive exudation of inflammatory materials and thickened pulmonary interstitium. The dexamethasone group and acupuncture group showed less damage of the alveolar structure and only a small number of inflammatory cells around the airway. Compared with the normal group, the apoptosis rate of EOS in lung tissue was decreased (<0.01), the expression levels of p-P38MAPK and ICAM-1 proteins and mRNAs in the lung tissue were up-regulated (<0.01), while the expression levels of IFN-γ protein and mRNA in the lung tissue were down-regulated (<0.01) in the model group. Compared with the model group, the apoptosis rate of EOS in the lung tissue was increased (<0.05), the expression levels of p-P38MAPK and ICAM-1 proteins and mRNAs in the lung tissue were down-regulated (<0.01, <0.05), while the expression levels of IFN-γ protein and mRNA were up-regulated (<0.05, <0.01) in the dexamethasone and acupuncture groups.
Acupuncture may inhibit the P38MAPK signaling pathway, down-regulate ICAM-1 expression, and up-regulate IFN-γ expression to promote the apoptosis of EOS and reduce EOS aggregation, thus alleviating the inflammatory response of airway in asthma.
观察针刺对哮喘大鼠肺组织磷酸化P38丝裂原活化蛋白激酶（p-P38MAPK）、细胞间黏附分子-1（ICAM-1）、干扰素γ（IFN-γ）及嗜酸性粒细胞（EOS）的影响，探讨针刺调控EOS凋亡的机制。
将清洁级雄性Wistar大鼠随机分为正常组、模型组、地塞米松组和针刺组，每组8只。采用腹腔注射10%卵白蛋白与10%氢氧化铝混合悬液（1 mL）加生理盐水的方法制备哮喘模型，随后雾化吸入1%卵白蛋白溶液30 min，每日1次，连续2周以诱发哮喘症状。地塞米松组自第15天起腹腔注射地塞米松0.9 mg/kg，每日1次，连续2周。针刺组自第15天起隔日选取双侧“肺俞”（BL13）、“脾俞”（BL20）、“肾俞”（BL23）、“定喘”（EX-B1）、“膻中”（CV17）进行针刺治疗，连续2周。行提插补泻手法，留针30 min。采用苏木精-伊红（HE）染色观察肺组织病理变化。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL法）检测大鼠肺组织EOS凋亡情况。采用蛋白质印迹法检测肺组织中p-P38MAPK的表达。分别采用免疫组化法和实时荧光定量PCR法检测肺组织中ICAM-1和IFN-γ的蛋白及mRNA水平。
HE染色结果显示，正常组肺泡及周围组织完整，无炎性细胞浸润。模型组可见大量炎性物质渗出，肺间质增厚。地塞米松组和针刺组肺泡结构损伤较轻，气道周围仅有少量炎性细胞。与正常组比较，模型组肺组织EOS凋亡率降低（<0.01），肺组织中p-P38MAPK、ICAM-1蛋白及mRNA表达水平上调（<0.01），而肺组织中IFN-γ蛋白及mRNA表达水平下调（<0.01）。与模型组比较，地塞米松组和针刺组肺组织EOS凋亡率升高（<0.05），肺组织中p-P38MAPK、ICAM-1蛋白及mRNA表达水平下调（<0.01,<0.05），而IFN-γ蛋白及mRNA表达水平上调（<0.05,<0.01）。
针刺可能通过抑制P38MAPK信号通路，下调ICAM-1表达，上调IFN-γ表达，促进EOS凋亡，减少EOS聚集，从而减轻哮喘气道炎症反应。
