基于MAPK信号通路探讨针刺蝶腭穴对变应性鼻炎大鼠鼻-脾轴协同抗炎的作用机制

Exploration on the mechanism of acupuncture at Die’e acupoint on the synergistic anti-inflammatory effect of nose-spleen axis based on MAPK signaling pathway in rats with allergic rhinitis

目的:观察针刺蝶腭穴对变应性鼻炎大鼠鼻-脾轴免疫炎症的影响,探究其神经-免疫调控作用机制。方法:将24只SD大鼠随机分为空白组、模型组、针刺组、假针刺组,每组6只。卵清蛋白诱导法建立变应性鼻炎大鼠模型。记录干预前后各组大鼠行为学积分情况,采用甲苯胺蓝染色法、HE染色法分别观察大鼠鼻黏膜、脾脏组织病理形态,ELISA法检测大鼠鼻腔灌洗液中P物质(SP)、神经肽Y(NPY)、卵清蛋白特异性IgE(OVA-sIgE)、白介素(IL)-1β、肿瘤坏死因子α(TNF-α)、IL-6、IL-10及干扰素γ(IFN-γ)含量;Western blot法检测大鼠鼻黏膜及脾脏组织中ERK、p-ERK、P38、p-P38、JNK、p-JNK蛋白表达。结果:与模型组比较,干预后针刺组行为学积分显著降低(P<0.01),鼻腔灌洗液中的SP、OVA-sIgE、IL-1β、TNF-α、IL-6含量显著降低(P<0.01),NPY、IFN-γ、IL-10含量显著升高(P<0.01),鼻黏膜及脾脏p-ERK、p-P38、p-JNK蛋白磷酸化水平显著下降(P<0.05,P<0.01)。结论:针刺蝶腭穴可调控神经递质SP、NPY的释放水平,并对变应性鼻炎大鼠鼻-脾轴发挥协同抗炎作用,其机制可能与MAPK信号通路有关。

Objective:To observe the effect of acupuncture at Die’e acupoint on the immunoinflammation of the nosespleen axis in rats with allergic rhinitis,and to investigate the mechanism of neural-immunoregulation.Methods:Twenty-four SD rats were randomly divided into blank group,model group,acupuncture group and sham-acupuncture group,six in each group.Ovalbumin induction method was used to establish a rat model of allergic rhinitis.Behavioral scores of rats in each group were recorded before and after the intervention.Toluidine blue staining and HE staining were used to observe the histopathological morphology of the nasal mucosa and spleen of rats,respectively;ELISA was used to detect the contents of sustance P(SP),neuropeptide Y(NPY),ovalbumin specific IgE(OVA-sIgE),interleukin(IL)-1β,tumor necrosis factorα(TNF-α),IL-6,IL-10,and inteferonγ(IFN-γ)in the nasal lavage fluid of rats;and Western blot was used to detect ERK,p-ERK,P38,p-P38,JNK,p-JNK protein expression in rat nasal mucosa and spleen tissue.Results:Compared with the model group,the behavioral scores of the acupuncture group were significantly reduced after the intervention(P<0.01),the levels of SP,OVA-sIgE,IL-1β,TNF-αand IL-6 in the nasal lavage fluid were reduced(P<0.01),and the levels of NPY,IFN-γand IL-10 were elevated(P<0.01),and the phosphorylation levels of p-ERK,p-P38,and p-JNK proteins in the nasal mucosa and spleen decreased(P<0.05,P<0.01).Conclusion:Acupuncture at Die’e acupoint can regulate the release levels of neurotransmitters SP and NPY,and exert a synergistic anti-inflammatory effect on the nose-spleen axis in rats with allergic rhinitis.The mechanism may be related to the MAPK signaling pathway.