芪丹益肺通窍颗粒对变应性鼻炎大鼠免疫功能的影响及机制

Effect of Qidan Yifei Tongqiao Granules on immune function in rats and its mechanism

目的探究芪丹益肺通窍颗粒对变应性鼻炎(AR)大鼠免疫功能的影响及对白细胞介素-33(IL-33)/生长刺激表达基因2(ST2)通路的调控作用。方法取SD大鼠分为正常对照组、模型组(卵白蛋白致敏)、中药组(芪丹益肺通窍颗粒,28 g/kg)、pcDNA-IL-33组(尾静脉注射含IL-33过表达重组蛋白载体)、pcDNA-NC组(尾静脉注射pcDNA-IL-33阴性对照载体)、中药+pcDNA-IL-33组,每组20只;建立AR模型。观察大鼠鼻部症状,通过行为症状评分表,对大鼠鼻部AR症状进行评分;采用ELISA检测试剂盒检测血清免疫球蛋白(IgE)、白细胞介素-8(IL-8)、趋化因子CC基序配体2(CCL2)水平;HE染色观察鼻黏膜组织病理变化;免疫组化法检测鼻黏膜组织IL-33、ST2阳性表达水平;流式细胞术检测辅助性T细胞1(Th1)/辅助性T细胞2(Th2);采用ELISA检测试剂盒检测鼻腔灌洗液中免疫因子如肿瘤坏死因子-β(TNF-β)、白细胞介素(IL-2、IL-6、IL-9)水平;Western blot法检测Th1转录因子(T-bet)、Th2转录因子(Gata3)、IL-33、髓样分化蛋白88(MyD88)、肿瘤坏死因子受体相关因子6(TRAF6)表达。结果模型组大鼠鼻部AR症状评分升高,鼻黏膜上皮脱落及炎性浸润等病理损伤加重,嗜酸性粒细胞数目及血清IgE、CCL2、IL-8水平升高、Th1/Th2比例降低,IL-33/ST2通路及其相关蛋白表达升高(P<0.05)。芪丹益肺通窍颗粒可改善大鼠鼻部AR症状评分及鼻黏膜组织损伤、上调Th1/Th2比例、抑制IgE及炎症因子水平(P<0.05)。中药+pcDNA-IL-33组可逆转中药组的上述作用。结论芪丹益肺通窍颗粒可能通过抑制IL-33/ST2通路激活,改善大鼠Th1/Th2免疫功能失衡,缓解大鼠AR症状。

Objective To explore the effect of Qidan Yifei Tongqiao Granules(QYTG)on the immune function of rats with allergic rhinitis(AR),and its regulation via interleukin-33(IL-33)/growth-stimulating expressed gene 2(ST2)pathway.Methods SD rats were divided into normal control,AR model(sensitization with ovalbumin),QYTG(28 g/kg),pcDNA-IL-33(tail vein injection containing IL-33 overexpression recombinant protein vector),pcDNA-NC(tail vein injection of pcDNA-IL-33 negative control vector),and QYTG+pcDNA-IL-33 groups,with 20 animals each.The nasal AR symptoms of rats were scored;the levels of serum immunoglobulin(IgE),interleukin-8(IL-8),chemokine CC motif ligand 2(CCL2)were detected with ELISA;the pathological changes of nasal mucosa were observed with HE staining;the expression of IL-33 and ST2 in nasal mucosa tissue were detected with immunohistochemistry;the ratios of Th1/Th2 were detected with flow cytometry;the levels of tumor necrosis factor-β(TNF-β),IL-2,IL-6,and IL-9 in nasal lavage fluid were detected with ELISA;the expression of Th1 transcription factor(T-bet),Th2 transcription factor(Gata3),IL-33,myeloid differentiation protein 88(MyD88),tumor necrosis factor receptor related factor 6(TRAF6)was detected with Western blot.Results In the model group,the nasal AR symptom score increased,and pathological damages such as epithelial shedding and inflammatory infiltration of the nasal mucosa increased,the number of eosinophils and serum IgE,CCL2,and IL-8 levels increased,the ratios of Th1/Th2 were reduced,the expression of IL-33/ST2 pathway and its related proteins increased(P<0.05).QYTG can improve the nasal AR symptom score and nasal mucosal tissue damage,up-regulate the ratios of Th1/Th2,and inhibit the levels of IgE and inflammatory factors(P<0.05).QYTG+pcDNA-IL-33 could reverse the above-mentioned effects.Conclusion Qidan Yifei Tongqiao Granules relieved AR symptoms of rats by inhibiting the activation of IL-33/ST2 pathway and regulating Th1/Th2 immune function imbalance.