Objective: To observe the effect of acupuncture at "Feishu" (BL 13) + "Dingchuan" (EX-B 1) and "Kongzui" (LU 6) + "Yuji" (LU 10) for the airway remodeling in asthma rats based on the transforming growth factor-β1 (TGF-β1)/ Smad family member 3 (Smad3) signaling pathway; and explore the efficacy difference between the two acupoint combinations.
Methods: Forty SPF male SD rats, aged 4 weeks, were randomly divided into a blank group (n = 10) and a modeling group (n = 30). The ovalbumin (OVA) sensitization method was used to establish asthma model in the modeling group. After successful model preparation, the rats of the modeling group were randomized into a model group, an acupuncture at "Feishu" (BL 13) + "Dingchuan" (EX-B 1) (AAF) group, and acupuncture at "Kongzui" (LU 6)+"Yuji" (LU 10) (AAK) group, with 10 rats in each one. Starting from day 15 of the experiment, 5 min after motivating, acupuncture was applied to "Feishu" (BL 13) + "Dingchuan" (EX-B 1) and "Kongzui" (LU 6)+"Yuji" (LU 10) in the AAF group and the AAK group respectively. The intervention was delivered for 30 min each time, once daily, lasting 3 weeks consecutively. Using lung function detector, the airway resistance (RL) and dynamic compliance (Cdyn) of the lungs were detected. The histomorphology of lung tissues was detected with HE staining and Masson staining, and the mRNA and protein expression of TGF-β1 and Smad3 in lung tissues was detected with the real-time PCR and Western blot methods.
Results: Compared with the blank group, RL was increased and Cdyn was decreased in the rats of the model group (P<0.01); and RL was reduced and Cdyn was increased in the AAF group and the AAK group when compared with those in the model group (P<0.01, P<0.05). The rats of the model group had bronchial lumen stenosis, inflammatory cell infiltration, collagen fibre hyperplasia and thickened smooth muscle in the lung tissues when compared with those in the blank group; and in comparison with the model group, all of the above morphological changes were attenuated in the AAF group and the AAK group. Besides, these morphological changes of the lung tissues were more alleviated in the AAF group when compared with those in the AAK group. In comparison with the blank group, the mRNA and protein expression of TGF-β1 and Smad3 of the lung tissues was increased in the model group (P<0.01), and it was reduced in the AAF group and the AAK group when compared with that in the model group (P<0.05, P<0.01). The mRNA expression of TGF-β1 and Smad3 was lower in the AAF group when compared with that in the AAK group (P<0.05).
Conclusion: Acupuncture at either "Feishu" (BL 13)+"Dingchuan" (EX-B 1) or "Kongzui" (LU 6)+"Yuji" (LU 10) reduces the airway remodeling in the rats with asthma, which may be related to the down-regulation of mRNA and protein expression of TGF-β1 and Smad3. The better efficacy is obtained with acupuncture at "Feishu" (BL 13)+"Dingchuan" (EX-B 1).
目的：基于转化生长因子β1（TGF-β1）/Smad家族成员3（Smad3）信号通路观察针刺双侧“肺俞”“定喘”和双侧“孔最”“鱼际”对哮喘大鼠气道重塑的影响，探讨两组腧穴配伍的疗效差异。方法：将40只4周龄SPF级雄性SD大鼠随机分为空白组10只、造模组30只。造模组大鼠采用卵清蛋白（OVA）致敏法复制哮喘模型，模型制备成功后将大鼠随机分为模型组、针刺“肺俞”“定喘”（AAF）组、针刺“孔最”“鱼际”（AAK）组，每组10只。实验第15天起，AAF组、AAK组每天于激发后5 min分别针刺双侧“肺俞”“定喘”及双侧“孔最”“鱼际”，每次30 min，每日1次，连续3周。应用肺功能检测仪检测大鼠气道阻力（RL）和肺顺应性（Cdyn），HE染色法及Masson染色法检测肺组织形态，实时荧光定量PCR法及Western blot法检测肺组织TGF-β1、Smad3 mRNA和蛋白表达。结果：与空白组比较，模型组大鼠RL升高、Cdyn降低（P<0.01）；与模型组比较，AAF组和AAK组RL降低、Cdyn升高（P<0.01，P<0.05）。与空白组比较，模型组大鼠肺组织出现支气管管腔狭窄、炎性细胞浸润、胶原纤维增生、平滑肌增厚；与模型组比较，AAF组和AAK组支气管管腔狭窄、炎性细胞浸润、胶原纤维增生及平滑肌增厚减轻；与AAK组比较，AAF组以上肺组织形态改变均减轻。与空白组比较，模型组肺组织TGF-β1、Smad3 mRNA和蛋白表达升高（P<0.01）；与模型组比较，AAF组和AAK组肺组织TGF-β1、Smad3 mRNA和蛋白表达降低（P<0.05，P<0.01）；与AAK组比较，AAF组大鼠TGF-β1、Smad3 mRNA表达降低（P<0.05）。结论：针刺“肺俞”“定喘”及“孔最”“鱼际”均可减轻哮喘大鼠气道重塑，可能与下调TGF-β1、Smad3 mRNA和蛋白表达有关；针刺“肺俞”“定喘”疗效更优。.
Keywords: TGF-β1/Smad3 signaling pathway; acupuncture; airway remodeling; asthma.