摘要
目的:观察针刺与艾灸肺俞穴对哮喘模型大鼠炎症反应和肠道短链脂肪酸(SCFAs)的影响。方法:将56只Sprague-Dawley大鼠随机分为正常组16只和造模组40只。应用卵清白蛋白(OVA)制备哮喘大鼠模型。分别取正常组和造模组大鼠各4只进行模型评价。模型制备成功后,剩余造模组大鼠被随机分为模型组、针刺组和艾灸组,每组12只。针刺组和艾灸组大鼠均取双侧肺俞穴,分别进行针刺与艾灸干预30 min。每次干预结束后1 h,给予1%OVA溶液雾化激发20 min,持续14 d。实验结束后,采用生化法检测大鼠外周血中炎性相关指标;瑞氏-吉姆萨染色法计数肺泡灌洗液(BALF)中炎症细胞含量;苏木素-伊红染色后光镜下观察肺组织形态学病理变化;实时荧光定量聚合酶链反应检测肺组织中相关炎症因子mRNA表达量;气相色谱-氢火焰离子化法检测粪便中SCFAs的含量。结果:模型组大鼠外周血中嗜酸性粒细胞(Eos)、中性粒细胞(Neu)及淋巴细胞(Lym),BALF中Eos及Neu的含量及肺组织中白介素(IL)-4、IL-5、IL-13、IL-33、胸腺基质淋巴细胞生成素(TSLP)mRNA表达均显著升高(P<0.05或P<0.01);肺组织结构严重损伤;粪便中乙酸、丙酸、异丁酸、丁酸、戊酸含量均显著降低(P<0.05或P<0.01)。与模型组相比,针刺组与艾灸组大鼠外周血中的Eos、Neu、Lym,BALF中Eos含量及肺组织中IL-4 mRNA和IL-5 mRNA表达均显著降低(P<0.05或P<0.01),肺组织病理损伤减轻,粪便中乙酸、丙酸、异丁酸及丁酸含量均显著升高(P<0.05或P<0.01);针刺组大鼠BALF中Lym,肺组织中IL-13 mRNA表达以及艾灸组大鼠肺组织中IL-33 mRNA和TSLP mRNA表达均显著降低(P<0.05或P<0.01);艾灸组大鼠粪便中戊酸含量显著升高(P<0.01)。与针刺组相比,艾灸组大鼠肺组织中IL-5 mRNA和IL-13 mRNA表达,以及粪便中乙酸含量显著升高(P<0.05或P<0.01)。结论:针刺和艾灸均能改善哮喘模型大鼠异常炎症,调节肠道SCFAs含量,但针刺对促炎因子尤其是IL-5和IL-13的抑制效果优于艾灸,而艾灸对肠道菌群代谢物SCFAs尤其是乙酸的调节优于针刺。
Objective:To observe the effects of acupuncture and moxibustion at Feishu(BL13)on inflammatory responses and intestinal short-chain fatty acids(SCFAs)in rats with asthma.Methods:Fifty-six Sprague-Dawley rats were randomly divided into a normal group(16 rats)and a modeling group(40 rats).Rats in the modeling group were subjected to establishing asthma models using ovalbumin(OVA).Model evaluation was conducted using 4 rats from each group.The remaining rats that successfully developed asthma were then randomly divided into a model group,an acupuncture group,and a moxibustion group,with 12 rats in each group.Rats in the acupuncture group received acupuncture treatments,and those in the moxibustion group received moxibustion treatments,both at Feishu(BL13)for 30 min.Following the treatments,the rats were exposed to atomization excitation with a 1%OVA solution for 20 min daily for 14 consecutive days.At the end of the experiment,inflammatory markers in the rats’peripheral blood were analyzed using a biochemical method.In addition,inflammatory cells in the bronchoalveolar lavage fluid(BALF)were counted using Wright-Giemsa staining.The lung tissue of rats was examined under a light microscope after staining with hematoxylin-eosin to observe morphological or pathological changes.Furthermore,real-time fluorescence quantitative polymerase chain reaction was utilized to measure the mRNA expression of inflammatory factors in the lung tissue.Lastly,the concentration of SCFAs in the rat’s feces was determined using gas chromatographyhydrogen flame ionization.Results:The levels of eosinophils(Eos),neutrophils(Neu),and lymphocytes(Lym)in the peripheral blood,as well as Eos and Neu in the BALF,and the expression of interleukin(IL)-4,IL-5,IL-13,IL-33,and thymic stromal lymphopoietin(TSLP)mRNAs in the lung tissue were all found to be significantly increased(P<0.05 or P<0.01);the lung tissue structure displayed severe injuries;the levels of acetic acid,propionic acid,isobutyric acid,butyric acid,and valeric acid in the feces decreased significantly in the model group(P<0.05 or P<0.01).Compared with the model group,the peripheral blood levels of Eos,Neu,and Lym,as well as Eos in the BALF,and the mRNA expression levels of IL-4 and IL-5 in the lung tissue decreased significantly in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).This reduction was accompanied by alleviated pathological damage in the lung tissue.Additionally,there were significant increases in the levels of acetic acid,propionic acid,isobutyric acid,and butyric acid in the feces in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).In the acupuncture group,the expression levels of Lym in the BALF and IL-13 mRNA in the lung tissue decreased significantly(P<0.05 or P<0.01).In the moxibustion group,the mRNA expression levels of IL-33 and TSLP in the lung tissue also reduced significantly(P<0.05 or P<0.01).Furthermore,the level of valeric acid in the feces increased notably in the moxibustion group(P<0.01).Compared with the acupuncture group,it was found that the mRNA levels of IL-5 and IL-13 in the lung tissue,as well as the acetic acid level in the feces,were significantly higher in the moxibustion group(P<0.05 or P<0.01).Conclusion:Both acupuncture and moxibustion were effective in reducing abnormal inflammation and regulating intestinal SCFAs in asthma model rats.Acupuncture demonstrated superiority in inhibiting pro-inflammatory factors,particularly IL-5 and IL-13,while moxibustion exhibited better regulation on intestinal metabolites SCFAs,especially acetic acid.
作者
任玲
周竞颖
来奕恬
屈艺卓
张国山
杨燕萍(翻译)
REN Ling;ZHOU Jingying;LAI Yitian;QU Yizhuo;ZHANG Guoshan;YANG Yanping(School of Acupuncture-moxibustion,Tuina and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,China;不详)
基金
湖南省自然科学基金项目,No.2021JJ30513
湖南省教育厅科学研究项目,No.20B444
国家自然科学基金项目,No.81603705
中国博士后科学基金项目,No.2107M612567
湖南省青年骨干教师培养对象项目,2021
湖南中医药大学研究生创新课题,No.2023CX79。
关键词
灸法
针刺疗法
穴
肺俞
哮喘
炎症
短链脂肪酸
大鼠
Moxibustion Therapy
Acupuncture Therapy
Point,Feishu(BL13)
Asthma
Inflammation
Short-chain Fatty Acids
Rats
