摘要
目的:观察电针"足三里"和"肺俞"对慢性阻塞性肺疾病(COPD)大鼠肺泡巨噬细胞M1极化的影响,探讨电针抗炎的可能机制。方法:将40只SD大鼠随机分为正常组、正常电针组、模型组和模型电针组,每组10只。采用单纯烟熏法复制COPD大鼠模型。正常电针组和模型电针组大鼠给予电针双侧"足三里"和"肺俞",每次30 min,隔日治疗1次,持续2周。于治疗结束后检测大鼠肺功能;HE染色法观察大鼠肺组织病理变化;ELISA法检测大鼠支气管肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)、诱导型一氧化氮合酶(iNOS)的含量;Western blot法和荧光定量PCR法分别检测肺泡巨噬细胞(AM)中M1极化标志物(CD86、iNOS)及信号通路蛋白髓样分化因子88(MyD88)、核因子-κB p65(NF-κB p65)的表达;免疫组织化学法观察肺组织中CD86的分布及免疫表达。结果:与正常组比较,模型组大鼠肺组织内炎性细胞浸润明显,肺泡壁增厚;用力肺活量(FVC)、第0.1秒用力呼气量(FEV0.1)、第0.3秒用力呼气量(FEV0.3)、FEV0.1占FVC之比(FEV0.1/FVC),FEV0.3占FVC之比(FEV0.3/FVC)均显著下降(P<0.01);BALF中TNF-α、iNOS含量及AM中CD86、iNOS、MyD88、NF-κB p65蛋白及mRNA表达明显升高(P<0.01),肺组织中CD86阳性表达明显升高(P<0.01)。与模型组比较,模型电针组的炎性细胞浸润及肺泡壁厚度均有改善;FVC、FEV0.1、FEV0.3、FEV0.1/FVC及FEV0.3/FVC均显著上升(P<0.05,P<0.01);BALF中TNF-α、iNOS含量及AM中CD86、iNOS、MyD88、NF-κB p65蛋白及mRNA表达明显下降(P<0.01,P<0.05),肺组织中CD86阳性表达明显降低(P<0.01)。结论:电针"足三里"和"肺俞"可以降低COPD大鼠肺内炎性反应,其机制可能与电针通过抑制MyD88/NF-κB p65信号通路从而抑制AM向M1型极化有关。
Objective To observe the effect of electroacupuncture(EA)at"Zusanli"(ST36)and"Feishu"(BL13)on M1 polarization of alveolar macrophages(AM)in rats with chronic obstructive pulmonary disease(COPD),so as to explore its anti-inflammatory mechanism underlying improvement of COPD.Methods Forty SD rats were randomly divided into normal and normal+EA,COPD model and COPD+EA groups(n=10 in each group).The COPD model was established by simple fumigation.EA(4 Hz/20 Hz,1 to 2 mA)was applied to bilateral ST36 and BL13 for 30 min,once every other day for 2 weeks.The pulmonary function including the forced vital capacity(FVC),forced expiratory volume in 0.1 and 0.3 s(FEV0.1,FEV0.3,FEV0.1/FVC,and FEV0.3/FVC)was detected by using a small animal respiratory function detector.Histopathological changes of the lung were displayed by H.E.staining.The contents of tumor necrosis factor-α(TNF-α)and induced nitric oxide synthase(iNOS)in the broncho alveolar lavage fluid(BALF)were assayed by ELISA.The expression of M1 polarization markers(CD86,iNOS),myeloid differentiation factor 88(MyD88)and nuclear factor-κB p65(NF-κB p65)in AM were detected by Western blot and quantitative real time-PCR,separately.The distribution and expression of CD86 in the lung were detected by immunohistochemistry.Results Following modeling,the levels of FVC,FEV0.1,FEV0.3,ratios of FVE0.1/FVC and FEV0.3/FVC were significantly decreased(P<0.01),while the contents of TNF-αand iNOS in the BALF,expression of CD86,iNOS,MyD88 and NF-κB p65 mRNAs and proteins in the AM,and CD86 immunoactivity in the lung were significantly increased in the model group relevant to the normal group(P<0.01).After the intervention,the decrease of the lung function and increase of the above-mentioned genes and proteins were all reversed in the COPD+EA group(P<0.05,P<0.01).Conclusion EA at ST36 and BL13 can reduce pulmonary inflammation in COPD rats,which may be related to its function in inhibiting M1 polarization of AM via down-regulating MyD88/NF-κB p65 signaling pathway.
作者
李尹
张新芳
刘自兵
詹婉婷
张毅
程晨
谢菁菁
LI Yin;ZHANG Xin-fang;LIU Zi-bing;ZHAN Wan-ting;ZHANG Yi;CHENG Chen;XIE Jing-jing(Graduate Schoo,Anhui University of Chinese Medicine,Hefei 230038,China;Department of Physiology,College of Integrated Traditional Chinese and Western Medicine,Anhui University of Chinese Medicine,Hefei 230038,China;Institute of Acupuncture and Meridian,College of Acupuncture-moxibustion and Massage,Anhui University of Chinese Medicine,Hefei 230038,China;Key Laboratory of Xin′an Medicine,Ministry of Education,Anhui University of Chinese Medicine,Hefei 230038,China)
出处
《针刺研究》
CAS
CSCD
北大核心
2020年第3期173-179,共7页
Acupuncture Research
基金
国家自然科学基金项目(No.81373743)
安徽省自然科学基金项目(No.1408085MH201)
安徽中医药大学研究生科技创新基金(No.2018YB008)。
作者简介
第一作者:李尹,2017级硕士研究生,研究方向:针刺作用机制。E-mail:3076247927@qq.com;通信作者:刘自兵,教授,硕士生导师,研究方向:针刺作用机制。E-mail:zibingliu@163.com。
