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[艾灸肺俞穴与肺募穴对哮喘小鼠Th17/Treg平衡的影响]

[Effect of moxa-cone moxibustion at lung's back- points and front- points on Th17/Treg balance in mice with asthma].

作者信息

Qiao Yun, Lei Hui-Ting, Yi Wei, Zhao Cai-Jiao, Xia Xing, Zhou Bei, Rui Jing-Lin, Wang Zeng-Duo

机构信息

College of Acupuncture-Moxibustion and Tuina, Guangxi University of CM, Nanning 530200, China.

College of Graduate, Guangxi University of CM, Nanning 530200, China.

出版信息

Zhongguo Zhen Jiu. 2020 Nov 12;40(11):1217-22. doi: 10.13703/j.0255-2930.20191004-0001.

Abstract

OBJECTIVE

To observe the effect of moxa-cone moxibustion at lung's back- points and front- 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 (<0.01), while that in the LY group, the EA group and the moxibustion group was longer than that in the model group (<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 (<0.01), while the content of IL-10 in serum and BALF and the percentage of Treg in spleen tissue were decreased (<0.01, <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 (<0.01, <0.05), while the content of IL-10 in serum and BALF and the percentage of Treg in spleen tissue were increased (<0.01, <0.05). The above indice had no significant difference among all the intervention groups (>0.05).

CONCLUSION

The Th17/Treg is imbalanced in asthmatic body. The moxibustion at lung's back- points and front- points can control asthma by regulating Th17/Treg imbalance and restoring immune homeostasis.

摘要

目的

观察艾灸肺俞穴与中府穴对哮喘小鼠辅助性T淋巴细胞17(Th17)/调节性T淋巴细胞(Treg)表达的影响,探讨艾灸治疗哮喘的可能机制。

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组)、电针组和艾灸组,每组12只。采用卵清蛋白(OVA)致敏法复制哮喘模型。除正常组小鼠外,其余小鼠于第1、7、14天腹腔注射致敏液(含15μgOVA和30mg氢氧化铝),每只0.5mL;从第15天起,用1%OVA溶液雾化20min,每天1次,共2周;正常组小鼠采用相同操作,但注射0.9%氯化钠溶液。LY组小鼠于第13、14、15天经尾静脉注射LY294002。第15天开始,电针组小鼠取“肺俞”(BL13)、“中府”(LU1)穴,采用疏密波,频率2Hz/20Hz,强度1mA,每次15min,每天1次,共2周。艾灸组小鼠从第15天起对“肺俞”(BL13)、“中府”(LU1)穴进行艾炷灸,每穴3壮,每天1次,共两周。于第16、18、22天记录哮喘潜伏期。第29天,采集所有样本。采用ELISA法检测血清及支气管肺泡灌洗液(BALF)中IL-17和IL-10的表达。采用HE染色观察肺组织病理变化。采用流式细胞术检测脾组织中Th17、Treg百分比及Th17/Treg比值。

结果

与正常组比较,模型组哮喘潜伏期明显缩短(<0.01),LY组、电针组和艾灸组哮喘潜伏期均长于模型组(<0.01)。干预后,模型组肺组织有出血和炎性渗出,支气管和肺泡周围有大量炎性细胞浸润,出现广泛实变;LY组、电针组和艾灸组肺组织病理改变明显减轻,支气管和肺泡结构基本正常,炎性渗出和炎性细胞浸润较模型组明显减少;艾灸组炎症反应较LY组和电针组稍轻。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Th17百分比及Th17/Treg比值升高(<0.01),血清及BALF中IL-10含量、脾组织中Treg百分比降低(<0.01,<0.05)。与模型组比较,LY组、电针组和艾灸组血清及BALF中IL-17含量、脾组织中Th17百分比及Th17/Treg比值明显降低(<0.01,<0.05),血清及BALF中IL-10含量、脾组织中Treg百分比升高(<0.01,<0.05)。各干预组上述指标比较差异无统计学意义(>0.05)。

结论

哮喘机体存在Th17/Treg失衡。艾灸肺俞穴与中府穴可通过调节Th17/Treg失衡、恢复免疫稳态而控制哮喘。

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