针刺研究

2021, v.46(09) 757-762

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针刺结合无烟/有烟艾灸调节慢性鼻-鼻窦炎小鼠免疫功能的研究
Acupuncture combined with smokeless or smoky moxibustion for regulating immune function of experimental chronic rhinosinusitis mice

朱鑫;李辉;朱天民;李璐;胡守亮;
ZHU Xin;LI Hui;ZHU Tian-min;LI Lu;HU Shou-liang;Department of Rehabilitation Medicine, The Affiliated Taizhou Hospital of Wenzhou Medical University;College of Basic Medicine, Chengdu University;School of Health and Rehabilitation, Chengdu University of Traditional Chinese Medicine;

摘要(Abstract):

目的:探讨艾烟对慢性鼻-鼻窦炎(CRS)小鼠针灸结合治疗效应的影响,以及针刺结合无烟/有烟艾灸对胸腺基质淋巴细胞生成素(TSLP)、垂体腺苷酸环化酶激活肽(PACAP)蛋白的调节作用。方法:将雄性C57BL/6J小鼠随机分为正常组、假手术组、模型组、西药组、针刺结合无烟艾灸组和针刺结合有烟艾灸组,每组20只。采用开放上颌窦置入带有肺炎链球菌的膨胀海绵制备CRS小鼠模型。正常组不造模,假手术组开放上颌窦后缝合皮肤切口,其余各组小鼠开放上颌窦后置入带菌的海绵。西药组予克拉霉素0.103 g·kg~(-1)·d~(-1)灌胃;针刺结合无烟艾灸组和针刺结合有烟艾灸组均针刺双侧"足三里""肾俞",留针30 min,透刺双侧"合谷",分别予无烟艾灸、有烟艾灸"关元""神阙"穴20 min,针刺与艾灸隔日交替进行;正常组、假手术组、模型组以0.9%氯化钠溶液灌胃(200μL/d)。各组持续治疗21 d。HE染色法观察各组小鼠鼻窦黏膜病理形态;酶联免疫吸附法、免疫组织化学法观察各组小鼠鼻窦黏膜中TSLP、PACAP蛋白的表达。结果:与正常组、假手术组比较,模型组出现明显炎性反应;西药组、针刺结合无烟艾灸组、针刺结合有烟艾灸组鼻部炎性反应程度均较模型组减轻,且针刺结合有烟艾灸组鼻部炎性反应程度改善明显。与正常组、假手术组比较,模型组鼻窦黏膜中PACAP蛋白表达降低(P<0.05),其余组间鼻窦黏膜中PACAP蛋白的表达差异无统计学意义(P>0.05)。模型组鼻窦黏膜中TSLP蛋白的表达高于正常组、假手术组(P<0.01);与模型组比较,针刺结合有烟艾灸组、西药组鼻窦黏膜中TSLP蛋白表达降低(P<0.01),针刺结合无烟艾灸组鼻窦黏膜中TSLP蛋白表达升高(P<0.01);针刺结合无烟艾灸组鼻窦黏膜中TSLP蛋白的表达高于针刺结合有烟艾灸组(P<0.01)。结论:CRS的病理机制可能与鼻窦黏膜中PACAP蛋白的下调、TSLP的上调有关;针刺结合有烟艾灸通过降低鼻窦黏膜中TSLP的表达,能减轻CRS小鼠炎性反应及组织结构的破坏。
Objective To observe the therapeutic effect of moxa fume in the treatment of chronic rhinosinusitis(CRS) and the effect of acupuncture plus smokeless moxibustion or smoky moxibustion on the expression of thymic stromal lymphopoietin(TSLP) and pituitary adenylate cyclase activating polypeptide(PACAP) proteins in the sinus mucosal tissue in CRS mice.Me-thods Sixty male C57 BL/6 J mice were randomly divided into 6 groups, namely normal control, sham operation, CRS model, medication, acupuncture plus smokeless moxibustion(Acu+smokeless Moxi) and acupuncture plus smoky moxibustion(Acu+smoky Moxi) groups, with 20 mice in each group. The CRS model was established by inserting a piece of polyporous sponge filled with streptococcus pneumoniae into the maxillary sinus after operation. The mice in the sham operation group received skin incision after opening the maxillary sinus. Mice of the medication group received gavage of clarithromycin 0.103 g·kg~(-1)·d~(-1) for 21 days. For mice of the Acu+smokeless Moxi and Acu +smoky Moxi groups, manual acupuncture stimulation was applied to bilateral "Zusanli"(ST36), "Shenshu"(BL23) and "Hegu"(LI4) with the needles retained for 30 min, once every other day, and on the following day, moxibustion was applied to "Guanyuan"(CV4) and "Shenque"(BL23) for 20 min, once every other day. The treatment was given for 21 days. Mice of the normal, sham operation and model groups received gavage of normal saline(200 μL/d) for 21 days. Histopathological changes of the nasal mucosa were observed after H.E. staining, the TSLP and PACAP contents and expression were determined by enzyme-linked immunosorbent assay(ELISA) and immunohistochemistry, separately.Results At the end of the treatment, mice of the model group still had symptoms of nasal obstruction and runny nose, but those of the 3 treatment groups were obviously relieved in the nasal symptoms. H.E. staining showed an obvious chronic inflammatory reaction in the sinus mucosa, uneven distribution of the mucosal epithelium and necrotic and exfoliated epithelial cells, hyperplasia of fibrous tissue in the submucosa, etc. in the model group, which were relatively milder in the medication, Acu+smokeless Moxi and Acu+smoky Moxi groups, while no obvious inflammation was found in the normal group and sham operation group. In comparison with the normal group, no significant changes were found in the expression levels of PACAP and TSLP in the sham operation group(P>0.05). The expression level of PACAP was significantly lower(P<0.05) and that of TSLP significantly higher in the model group than in the normal and sham operatin groups(P<0.05). Compared with the model group, no significant changes were found in the expression of PACAP in the medication, Acu+smokeless Moxi and Acu+smoky Moxi groups(P>0.05), and the expression of TSLP was further obviously increased in the Acu+smokeless Moxi group(P<0.01), but obviously decreased in the Acu+smoky Moxi group(P<0.01).Conclusion Acupuncture combined with smoky moxibustion can down-regulate the expression of TSLP protein in the nasal sinus mucosa in CRS mice, which maybe contribute to its effect in reducing the inflammatory reaction and nasal symptoms.

关键词(KeyWords): 慢性鼻-鼻窦炎;针灸;垂体腺苷酸环化酶激活肽;胸腺基质淋巴细胞生成素
Chronic rhinosinusitis;Acupuncture plus moxibustion;Pituitary adenylate cyclase activating polypeptide;Thymic stromal lymphopoietin

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81674037);; 四川省科技计划项目(No.2018SZ0375)

作者(Author): 朱鑫;李辉;朱天民;李璐;胡守亮;
ZHU Xin;LI Hui;ZHU Tian-min;LI Lu;HU Shou-liang;Department of Rehabilitation Medicine, The Affiliated Taizhou Hospital of Wenzhou Medical University;College of Basic Medicine, Chengdu University;School of Health and Rehabilitation, Chengdu University of Traditional Chinese Medicine;

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DOI: 10.13702/j.1000-0607.201021

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