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[艾灸腧募穴对哮喘小鼠RORγt/Foxp3表达的干预机制:基于PI3K信号通路的分析]

[Intervention mechanism of moxa-cone moxibustion at Shu- and Mu-acupoints on the expression of RORγt/Foxp3 in asthma mice: an analysis based on PI3K signaling pathway].

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 and Massage, Guangxi University of Chinese Medicine, Nanning 530200, China.

Graduate School, Guangxi University of Chinese Medicine, Nanning 530200, China.

出版信息

Zhen Ci Yan Jiu. 2021 Apr 25;46(4):272-7. doi: 10.13702/j.1000-0607.200491.

Abstract

OBJECTIVE

To explore the effect of moxa-cone moxibustion at "Feishu" (BL13) and "Zhongfu" (LU1) on the contents of related inflammatory factors in serum and bronchoalveolar lavage fluid (BALF), and the expression of phosphatidy-linositol-3-kinase (PI3K), retinoic acid related orphan receptor γt (RORγt) and fork/wing helix transcription factor 3 (Foxp3) in lung tissue in asthma mice.

METHODS

Sixty male Balb/c mice were divided into normal, model, LY294002 (an inhibitor of PI3K, LY), electroacupuncture (EA) and moxa-cone moxibustion (moxibustion) groups (=12 in each group). The asthma model was established by intraperitoneal injection of ovalbumin sensitization. The mice in the LY group were injected with LY 294002 (7.5 mg/kg) via the tail vein. EA or moxa-cone moxibustion was applied at BL13 and LU1 in the EA or moxibustion group once daily for 2 weeks. The levels of immunoglobulin E (IgE), interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and BALF were detected by enzyme-linked immunosorbent assay. The expressions of PI3K, RORγt and Foxp3 in lung tissue were detected by real-time fluorescent quantitative polymerase chain reaction (QRT PCR) and immunohistochemistry.

RESULTS

Compared with the normal group, the levels of IgE and IL-4 in serum and BALF, the expressions of PI3K and RORγt mRNA and protein in lung tissue were significantly increased (<0.01), while the levels of IFN-γ, and the expressions of Foxp3 mRNA and protein were significantly reduced (<0.01) in the model group. Compared with the model group, the levels of IgE and IL-4 in serum and BALF, the expressions of PI3K and RORγt mRNA and protein in lung tissue were significantly decreased (<0.01, <0.05), and the levels of IFN-γ in serum and BALF, the expressions of Foxp3 mRNA and protein in lung tissue were significantly increased (<0.01) in the LY, EA and moxibustion groups.

CONCLUSION

Moxa cone moxibustion at Shu- and Mu-acupoints of the lung meridian can reduce airway inflammatory reaction and control asthma attacks in asthma mice, which is closely related to its effects in regulating the expressions of RORγt and Foxp3 through PI3K signaling pathway.

摘要

目的

探讨艾灸肺俞穴(BL13)、中府穴(LU1)对哮喘小鼠血清及支气管肺泡灌洗液(BALF)中相关炎症因子含量,以及肺组织中磷脂酰肌醇-3激酶(PI3K)、维甲酸相关孤儿受体γt(RORγt)和叉头/翼状螺旋转录因子3(Foxp3)表达的影响。

方法

将60只雄性Balb/c小鼠分为正常组、模型组、LY294002组(PI3K抑制剂LY)、电针组(EA)和艾灸组(每组12只)。采用腹腔注射卵白蛋白致敏法建立哮喘模型。LY组小鼠经尾静脉注射LY294002(7.5mg/kg)。EA组或艾灸组每日1次对BL13和LU1进行电针或艾灸,共2周。采用酶联免疫吸附测定法检测血清和BALF中免疫球蛋白E(IgE)、白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)水平。采用实时荧光定量聚合酶链反应(QRT-PCR)和免疫组织化学法检测肺组织中PI3K、RORγt和Foxp3的表达。

结果

与正常组比较,模型组血清和BALF中IgE、IL-4水平,肺组织中PI3K、RORγt mRNA及蛋白表达显著升高(P<0.01),而血清和BALF中IFN-γ水平,肺组织中Foxp3 mRNA及蛋白表达显著降低(P<0.01)。与模型组比较,LY组、EA组和艾灸组血清和BALF中IgE、IL-4水平,肺组织中PI3K、RORγt mRNA及蛋白表达显著降低(P<0.01,P <0.05),血清和BALF中IFN-γ水平,肺组织中Foxp3 mRNA及蛋白表达显著升高(P<0.01)。

结论

艾灸肺经俞募穴可减轻哮喘小鼠气道炎症反应,控制哮喘发作,这与其通过PI3K信号通路调节RORγt和Foxp3表达的作用密切相关。

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