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Title: [Effect of moxa-cone moxibustion at lung's back-shu points and front-mu points on Th17/Treg balance in mice with asthma].
Author: Qiao Y, Lei HT, Yi W, Zhao CJ, Xia X, Zhou B, Rui JL, Wang ZD.
Journal: Zhongguo Zhen Jiu; 2020 Nov 12; 40(11):1217-22. PubMed ID: 33788491.
Abstract:
OBJECTIVE: To observe the effect of moxa-cone moxibustion at lung's back-shu points and front-mu points on the expression of helper T lymphocyte 17 (Th 17)/regulatory T lymphocyte (Treg) in mice with asthma, and to explore the possible mechanism of moxa-cone moxibustion on asthma. METHODS: Sixty SPF-grade healthy male Balb/c mice were randomly divided into a normal group, a model group, an LY294002 group (LY group), an electroacupuncture (EA) group and a moxibustion group, 12 mice in each group. Asthma model was replicated by using ovalbumin (OVA) sensitization. Except the mice in the normal group, all the mice were intraperitoneally injected with sensitization solution (containing 15 μg of OVA and 30 mg of aluminum hydroxide) on the 1st day, 7th day and 14th day, 0.5 mL per mice; from the 15th day, 1% OVA solution was atomized for 20 min, once a day for 2 weeks; the mice in the normal group was treated with identical operations but with 0.9% sodium chloride solution. The mice in the LY group were treated with injection of LY294002 at tail vein on the 13th day, 14th day and 15th day. At the beginning of the 15th day, The mice in the EA group were treated with EA at "Feishu" (BL 13) and "Zhongfu" (LU 1) with disperse-dense wave, frequency of 2 Hz/20 Hz, intensity of 1 mA, 15 min each time, once a day for 2 weeks. The mice in the moxibustion group was treated with moxa-cone moxibustion at "Feishu" (BL 13) and "Zhongfu" (LU 1) from the 15th day, three moxa-cones per acupoint, once a day for 2 weeks. On the 16th day, 18th day and 22nd day, the incubation period of asthma was recorded. On the 29th day, all the samples were collected. The expressions of IL-17 and IL-10 in serum and bronchoalveolar lavage fluid (BALF) were detected by ELISA method. The pathological changes of lung tissue were observed by HE staining. The percentage of Th17, Treg and Th17/Treg ratio in spleen tissue were detected by flow cytometry method. RESULTS: Compared with the normal group, the incubation period of asthma in the model group was significantly shortened (P<0.01), while that in the LY group, the EA group and the moxibustion group was longer than that in the model group (P<0.01). After the intervention, bleeding and inflammatory exudation in the lung tissue were observed in the model group, and a large number of inflammatory cell infiltration around the bronchus and alveoli was found, and extensive consolidation appeared; the pathological changes of lung tissue in the LY group, the EA group and the moxibustion group were significantly reduced, and the structure of bronchus and alveoli was basically normal, and the inflammatory exudation and inflammatory cell infiltration were significantly reduced compared with the model group; the inflammatory reaction in the moxibustion group was slightly less than that in the LY group and the EA group. Compared with the normal group, the content of IL-17 in serum and BALF, the percentage of Th17 and Th17/Treg ratio in spleen tissue in the model group after intervention were increased (P<0.01), while the content of IL-10 in serum and BALF and the percentage of Treg in spleen tissue were decreased (P<0.01, P<0.05). Compared with the model group, the contents of IL-17 in serum and BALF, Th17 percentage and Th17/Treg ratio in spleen tissue in the LY group, the EA group and the moxibustion group were significantly reduced (P<0.01, P<0.05), while the content of IL-10 in serum and BALF and the percentage of Treg in spleen tissue were increased (P<0.01, P<0.05). The above indice had no significant difference among all the intervention groups (P>0.05). CONCLUSION: The Th17/Treg is imbalanced in asthmatic body. The moxibustion at lung's back-shu points and front-mu points can control asthma by regulating Th17/Treg imbalance and restoring immune homeostasis. 目的：观察肺“俞募配穴”艾炷灸对哮喘小鼠辅助性T淋巴细胞17（Th17）/调节性T淋巴细胞（Treg）表达的影响，探讨艾炷灸治疗哮喘的可能作用机制。方法：将60只SPF级健康雄性Balb/c小鼠随机分为正常组、模型组、LY294002组（LY剂组）、电针组和艾炷灸组，各12只，采用卵清蛋白（OVA）致敏复制哮喘模型。除正常组外，其他组小鼠于实验第1、7、14天腹腔注射以0.9%氯化钠溶液配制的致敏液（含OVA 15μg、氢氧化铝30 mg）0.5 mL/只，第15天起予1%OVA溶液雾化激发，每次20 min，每天1次，连续2周；正常组以0.9%氯化钠溶液代替进行以上操作。LY剂组于实验第13、14、15天予尾静脉注射LY294002；电针组于第15天起予电针“肺俞”“中府”穴，疏密波，频率2 Hz/20 Hz，强度1 mA，每次15 min，每日1次，连续2周；艾炷灸组于第15天起予艾炷灸“肺俞”和“中府”穴，每次每穴灸3壮，每日1次，连续2周。每组小鼠于实验第16、18、22天记录引喘潜伏期，第29天统一取材，予ELISA法检测血清及肺泡灌洗液（BALF）中IL-17和IL-10表达，HE染色法观察肺组织病理形态改变，流式细胞术检测脾组织Th17、Treg百分比和Th17/Treg比值。结果：与正常组比较，模型组小鼠引喘潜伏期明显缩短（P<0.01），LY剂组、电针组和艾炷灸组小鼠引喘潜伏期较模型组延长（P<0.01）。干预后，模型组小鼠肺组织内见出血及大量炎性渗出，支气管及肺泡周围大量炎性细胞浸润，出现广泛的实变；LY剂组、电针组及艾炷灸组的肺组织病变程度明显减轻，支气管及肺泡结构基本正常，炎性渗出及炎性细胞浸润较模型组明显减少，艾炷灸组的炎性反应稍轻于LY剂组和电针组。与正常组比较，干预后模型组血清及BALF中IL-17含量，脾组织中Th17细胞百分比、Th17/Treg比值升高（P<0.01），血清及BALF中IL-10含量、脾组织中Treg细胞百分比下降（P<0.01，P<0.05）；与模型组比较，LY剂组、电针组和艾炷灸组血清及BALF中IL-17含量，脾组织中Th17细胞百分比、Th17/Treg比值下降（P<0.05，P<0.01），血清及BALF中IL-10含量、脾组织中Treg细胞百分比升高（P<0.05，P<0.01）。各干预组间以上指标比较，差异均无统计学意义（P>0.05）。结论：哮喘机体存在Th17/Treg失衡，肺“俞募配穴”艾炷灸疗法能通过调控Th17/Treg失衡，使机体免疫恢复稳态而达到控制哮喘的目的。.

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