College of Acupuncture-Moxibustion and Tuina, Guangxi University of CM, Nanning 530200, China.
Basic Medicine School, Guangxi University of CM, Nanning 530200, China.
Zhongguo Zhen Jiu. 2024 Mar 12;44(3):295-302. doi: 10.13703/j.0255-2930.20230426-k0004.
To explore the effect and mechanism of acupuncture at "Feishu" (BL 13) and "Dingchuan" (EX-B 1), and "Kongzui" (LU 6) and "Yuji" (LU 10) for relaxing the airway smooth muscle in the rats during acute asthma attack and compare the effect among the two pairs of acupoints and the acupoints combination.
Forty SD male rats with SPF grade were randomly divided into a blank group, a model group, a pair-point A group (acupuncture at "Feishu" [BL 13] and "Dingchuan" [EX-B 1]), a pair-point B group (acupuncture at "Kongzui" [LU 6] and "Yuji" [LU 10]) and a point combination group (acupuncture at "Feishu" [BL 13] , "Dingchuan" [EX-B 1], "Kongzui" [LU 6] and "Yuji" [LU 10]), with 8 rats in each group. Except the rats in the blank group, the model of acute asthma attack was induced by ovalbumin (OVA) combined with aluminum hydroxide gel in the rest groups. Started on the 15th day of modeling, except in the blank group and the model group, acupuncture was delivered in the other groups, 30 min in each intervention, once daily, for 14 days. In each group, the latent period of asthma inducing was measured; the lung resistance (LR) and dynamic lung compliance (Cdyn) were determined using lung function detector; the levels of endothelin-1 (ET-1), tumor necrosis factor-α (TNF-α), cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in serum and bronchoalveolar lavage fluid (BALF) were measured by ELISA; with Masson staining and electron microscopy adopted, the morphology and ultrastructure of airway smooth muscle of the rats were observed; the mRNA and protein expressions of ET-1 and beta-2 adrenergic receptor (β-AR) were detected by quantitative real-time fluorescence and Western blot, respectively.
Compared with the blank group, the latent period of asthma inducing was shortened (0.05), RL increased and Cdyn decreased (0.05) with the different concentrations of methacholine (0.025 mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.2 mg/kg) in the model group. In the pair-point A group, the pair-point B group and the point combination group, the latent period of asthma inducing was prolonged (0.05), RL decreased and Cdyn increased (0.05) with different concentrations of methacholine when compared with those in the model group; and the latent period of asthma inducing in the point combination group was longer than that in the pair-point A group (<0.05). Compared with the blank group, the levels of ET-1, TNF-α and cGMP in the serum and BALF were elevated (<0.05), and those of cAMP reduced (0.05) in the model group. The levels of ET-1, TNF-α and cGMP in the serum and BALF were reduced (0.05), and those of cAMP elevated (<0.05) in the pair-point A group, the pair-point B group and the point combination group when compared with those in the model group. In the blank group, the lung tissue was normal structurally. In the model group, the collagen fibers were proliferated increasingly, the smooth muscle was thickened, the mitochondria were swollen, and their cristae disrupted and reduced massively. In the pair-point B group, the collagen fibers were proliferated, the smooth muscle was thicker compared with that in the blank group, the mitochondria were mildly swollen and their cristae disrupted partially. In the pair-point A group and the point combination group, the lung tissue changes were obviously alleviated in comparison with the model group, the mitochondria were slightly swollen and their cristae disrupted occasionally. Compared with the blank group, the mRNA and protein expression of ET-1 increased and that of β-AR decreased in the lung tissue of the model group (0.05). In the pair-point A group, the pair-point B group and the point combination group, the mRNA and protein expression of ET-1 was reduced and that of β-AR elevated in the lung tissue when compared with those in the model group (0.05). In comparison with the pair-point A group, the mRNA expression of β-AR was elevated in the point combination group (0.05). When compared with the pair-point B group, the mRNA expression of β-AR increased, the protein expression of ET-1 decreased (<0.05) in the point combination group.
Acupuncture at "Feishu" (BL 13) and "Dingchuan" (EX-B 1), "Kongzui" (LU 6) and "Yuji" (LU 10), two pairs of acupoints relieves the airway smooth muscle spasm in the rats during acute asthma attack, which may be related to inhibiting the mRNA and protein expression of ET-1 to reduce the excretion of ET-1 and TNF-α; while enhancing the mRNA and protein expression of β-AR to balance the levels of cAMP and cGMP. The effect is optimal when acupuncture is delivered at two pairs of acupoints simultaneously.
探讨针刺“肺俞”（BL 13）、“定喘”（EX-B 1）、“孔最”（LU 6）、“鱼际”（LU 10）对急性哮喘发作大鼠气道平滑肌松弛作用，并比较两对穴位和穴位组合的作用。
40 只 SPF 级雄性 SD 大鼠随机分为空白组、模型组、对穴 A 组（针刺“肺俞”[BL 13]、“定喘”[EX-B 1]）、对穴 B 组（针刺“孔最”[LU 6]、“鱼际”[LU 10]）和穴位组合组（针刺“肺俞”[BL 13]、“定喘”[EX-B 1]、“孔最”[LU 6]、“鱼际”[LU 10]），每组 8 只。除空白组外，其余各组大鼠均用卵清蛋白（OVA）联合氢氧化铝凝胶诱导急性哮喘模型。造模第 15 天起，除空白组和模型组外，其余各组大鼠进行针刺干预，每日 1 次，每次 30 min，共 14 天。测量各组大鼠哮喘诱发潜伏期；采用肺功能检测仪测定肺阻力（LR）和动态肺顺应性（Cdyn）；采用 ELISA 法检测各组大鼠血清和支气管肺泡灌洗液（BALF）中内皮素-1（ET-1）、肿瘤坏死因子-α（TNF-α）、环磷酸腺苷（cAMP）和环磷酸鸟苷（cGMP）水平；采用 Masson 染色和电镜观察大鼠气道平滑肌形态和超微结构；采用实时荧光定量 PCR 和 Western blot 法检测各组大鼠 ET-1 和β-2 肾上腺素能受体（β-AR）mRNA 和蛋白表达。
与空白组比较，模型组大鼠不同浓度（0.025、0.05、0.1、0.2 mg/kg）的乙酰甲胆碱诱导的哮喘潜伏期缩短（0.05），RL 增加，Cdyn 降低（0.05）。与模型组比较，对穴 A 组、对穴 B 组和穴位组合组大鼠不同浓度乙酰甲胆碱诱导的哮喘潜伏期延长（0.05），RL 降低，Cdyn 升高（0.05）；穴位组合组大鼠哮喘潜伏期长于对穴 A 组（<0.05）。与空白组比较，模型组大鼠血清和 BALF 中 ET-1、TNF-α和 cGMP 水平升高（<0.05），cAMP 水平降低（0.05）；与模型组比较，对穴 A 组、对穴 B 组和穴位组合组大鼠血清和 BALF 中 ET-1、TNF-α和 cGMP 水平降低（<0.05），cAMP 水平升高（<0.05）。空白组大鼠肺组织结构正常。模型组大鼠肺组织胶原纤维增生，平滑肌增厚，线粒体肿胀，嵴断裂减少。对穴 B 组大鼠肺组织胶原纤维增生，与空白组比较，平滑肌增厚，线粒体轻度肿胀，嵴部分断裂。对穴 A 组和穴位组合组大鼠肺组织改变明显减轻，线粒体轻度肿胀，嵴偶尔断裂。与空白组比较，模型组大鼠肺组织 ET-1 mRNA 和蛋白表达增加，β-AR 表达减少（0.05）；与模型组比较，对穴 A 组、对穴 B 组和穴位组合组大鼠肺组织 ET-1 mRNA 和蛋白表达降低，β-AR 表达升高（0.05）。与对穴 A 组比较，穴位组合组大鼠肺组织β-AR 表达升高（0.05）；与对穴 B 组比较，穴位组合组大鼠肺组织β-AR 表达升高，ET-1 蛋白表达降低（<0.05）。
针刺“肺俞”（BL 13）、“定喘”（EX-B 1）、“孔最”（LU 6）、“鱼际”（LU 10）两对穴位均可缓解急性哮喘发作大鼠气道平滑肌痉挛，其机制可能与抑制 ET-1 的 mRNA 和蛋白表达，减少 ET-1 和 TNF-α的释放，同时增强β-AR 的 mRNA 和蛋白表达，从而平衡 cAMP 和 cGMP 水平有关。两穴同时针刺作用最佳。
