鼻内针治疗变应性鼻炎大鼠及调节TH1/TH2免疫平衡的实验研究

Experimental study on treating allergic rhinitis(AR)rats with intranasal acupuncture and regulating TH1/TH2 immune balance

目的探讨鼻内针刺对变应性鼻炎(AR)大鼠的治疗效果,及对Th1/Th2免疫平衡的影响。方法选取20只SPF级SD雄性大鼠为研究对象,5只作为空白组,其余15只建立AR模型。造模成功后,随机分为模型组、针刺组、假针刺组,各5只。空白组和模型组不予干预,针刺组大鼠在迎香穴和鼻丘进行针刺治疗,假针刺组在面部非穴位区域浅刺。干预10 d后采用酶联免疫吸附试验(ELISA)检测血清中干扰素-γ(IFN-γ)和白细胞介素-4(IL-4)水平,苏木精-伊红(HE)染色观察鼻黏膜组织形态变化,蛋白质免疫印迹(WB)和实时荧光定量PCR(qPCR)分别检测鼻黏膜组织中Th1/Th2相关蛋白和基因的表达水平。结果与空白组比较,模型组行为评分、IL-4和GATA3、JAK1、STAT6的mRNA和蛋白水平均升高,IFN-γ、T-bet的mRNA和蛋白表达水平均降低,差异均有统计学意义(P<0.05)。与模型组比较,针刺组行为评分、IL-4和GATA3、JAK1、STAT6的mRNA和蛋白表达水平均降低,IFN-γ、T盒子转录因子(T-bet)的mRNA和蛋白表达水平均升高,差异均有统计学意义(P<0.05),且针刺组的行为评分、IL-4和GATA3、JAK1、STAT6的mRNA和蛋白表达水平均低于假针刺组,IFN-γ、T-bet的mRNA和蛋白表达水平均高于假针刺组,差异有统计学意义(P<0.05)。HE染色显示,针刺组大鼠鼻黏膜组织结构恢复接近正常,炎性细胞浸润显著减轻。结论鼻内针刺可能通过调节Th1/Th2免疫平衡而缓解AR症状。

ObjectiveExploring the therapeutic effect of intranasal acupuncture on allergic rhinitis(AR)rats and its impact on Th1/Th2 immune balance.Methods20 specific pathogen-free(SPF)SD male rats were selected for the study.5 rats were designated as control group,and the remaining 15 rats were used to establish an AR model.After successful modeling,the rats were randomly divided into 3 groups:model group,acupuncture group and sham acupuncture group,with 5 rats in each group.The blank group and model group received no intervention.The acupuncture group underwent acupuncture treatment at bilateral Yingxiang(LI20)and nasal bulb points,while the sham acupuncture group received superficial needling at non-acupoint,non-meridian locations.After 10 days of intervention,ELISA was used to detect the levels of interferon-gamma(IFN-γ)and interleukin-4(IL-4).Morphological changes in nasal mucosa tissues were observed via hematoxylin and eosin(HE)staining,while Th1/Th2-related protein and gene expressions were analyzed using Western blotting and quantitative PCR(qPCR),respectively.ResultsCompared with the blank group,the behavior score,IL-4,GATA3,JAK1,and STAT6 mRNA and protein levels in the model group increased,while the mRNA and protein expression levels of IFN-γand T-bet decreased(P<0.05).Compared with the model group,the behavioral scores,IL-4,GATA3,JAK1,and STAT6 mRNA and protein expression levels in the acupuncture group decreased,while the mRNA and protein expression levels of IFN-γand T-bet increased(P<0.05).The behavioral score,mRNA and protein expression levels of IL-4 and GATA3,JAK1,and STAT6 in the acupuncture group were lower than those in the sham acupuncture group,while the mRNA and protein expression levels of IFN-γand T-bet in the acupuncture group were higher than those in the sham acupuncture group(P<0.05).Hematoxylin and eosin(HE)staining revealed that the nasal mucosa tissue structure in the acupuncture group showed a near-normal appearance,with a significant reduction in inflammatory cell infiltration.ConclusionNasal acupuncture may alleviate AR symptoms by regulating Th1/Th2 immune balance.