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山东大学学报 (医学版) ›› 2020, Vol. 58 ›› Issue (1): 13-19.doi: 10.6040/j.issn.1671-7554.0.2019.339

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鼻腔滴入γ干扰素对变应性鼻炎大鼠外周血Th17/Treg细胞及相关细胞因子的影响

张美玲1,孙卉1,孙文凯2,孟朝暾3,邬信芳2,高伟4,李钦5   

  1. 1. 潍坊医学院临床医学院, 山东 潍坊 261000;2. 滨州医学院临床医学院, 山东 滨州 256600;3. 青岛大学医学部, 山东 青岛 266000;4. 山东第一医科大学研究生院, 山东 泰安 271000;5. 临沂市人民医院耳鼻咽喉头颈外科, 山东 临沂 276000
  • 发布日期:2022-09-27
  • 通讯作者: 李钦. E-mail:liqin8226@163.com
  • 基金资助:
    临沂市人民医院研究生培养基金(YJS2018047)

Effects of intranasal infusion of IFN-γ on Th17/Treg cells in peripheral blood of rats with allergic rhinitis

ZHANG Meiling1, SUN Hui1, SUN Wenkai2, MENG Zhaotun3, WU Xinfang2, GAO Wei4, LI Qin5   

  1. 1. Weifang Clinical Medical College, Weifang Medical University, Weifang 261000, Shandong, China;
    〓山〓东〓大〓学〓学〓报〓(医〓学〓版)58卷1期〓-张美玲, 等.鼻腔滴入γ干扰素对变应性鼻炎大鼠外周血Th17/Treg细胞及相关细胞因子的影响〓\=-2. School of Clinical Medicine, Binzhou Medical University, Binzhou 256600, Shandong, China;
    3. Medical College, Qingdao University, Qingdao 266000, Shandong, China;
    4. Graduate School, Shandong First Medical University, Taian 271000, Shandong, China;
    5. Department of Otorhinolaryngology &
    Head and Neck Surgery, Linyi Peoples Hospital, Linyi 276000, Shandong, China
  • Published:2022-09-27

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摘要： 目的 探讨大鼠鼻腔滴入γ干扰素(IFN-γ)对变应性鼻炎(AR)大鼠外周血中辅助性T细胞17(Th17)、调节性T细胞(Treg)水平的影响。 方法 将40只Wistar雄性大鼠,按体质量编号、采用随机数字表法随机分为4组:对照组、致敏组、曲安奈德组、IFN-γ组,致敏阶段采用5级卵清蛋白(OVA)、氢氧化铝给予致敏组、曲安奈德组、IFN-γ组大鼠腹腔注射致敏以及OVA滴鼻激发造模,对照组腹腔注射以生理盐水替代,滴鼻以超纯水替代;给药阶段对照组不予处理,致敏组、曲安奈德组、IFN-γ组每只每侧鼻孔分别给予50 μL PBS、3.5 μg曲安奈德、1 μg IFN-γ滴鼻,于末次给药结束24 h内行流式细胞术检测外周血Th17细胞、Treg细胞所占比例,1周内行定量酶联免疫吸附法(ELISA)检测外周血中白介素-17A、转化生长因子(TGF)-β1的浓度,并行苏木精-伊红(HE)染色,观察黏膜情况。 结果 与对照组［(0.213 4±0.149 5)%］相比,致敏组［(4.975 7±1.503 9)%］、曲安奈德组［(1.169 3±0.386 1)%］、IFN-γ组［(1.532 0±0.526 5)%］的Th17细胞比例都不同程度升高(F=51.532,P<0.05),除曲安奈德组、IFN-γ组间比较无统计学意义(P>0.05)外,其余各组间比较有统计学意义(P<0.05);与对照组［(22.939 3±2.099 5)%］相比,致敏组［(8.223 2±2.188 8)%］、曲安奈德组［(14.087 5±1.369 6)%］、IFN-γ组［(16.614 3±1.621 70)%］的Treg细胞比例不同程度降低(F=97.107,P<0.05),除曲安奈德组、IFN-γ组间比较无统计学意义(P>0.05)外,其余各组间比较有统计学意义(P<0.0083);与对照组［(5.076 9±1.435 6)pg/mL］相比,致敏组［(34.900 5±11.670 8)pg/mL］、曲安奈德组［(9.733 8±1.828 9)pg/mL］、IFN-γ组［(10.524 6±2.118 0)pg/mL］的IL-17A浓度升高(F=27.173,P<0.05),除曲安奈德组、IFN-γ组间比较无统计学意义(P>0.05)外,其余各组间比较有统计学意义(P<0.05);与对照组［(431.881 6±67.326 2)pg/mL］相比,致敏组［(173.886 2±41.708 2)pg/mL］、曲安奈德组［(246.053 3±13.841 2)pg/mL］、IFN-γ组［(290.012 7±22.885 6)pg/mL］的TGF-β1浓度降低(F=36.623,P<0.05),各组间比较有统计学意义(P<0.05)。与对照组大鼠黏膜相比,致敏组大鼠黏膜增厚、水肿,有大量炎性细胞浸润,细胞排列紊乱,杯状细胞增生,纤毛几乎不可见;曲安奈德组大鼠鼻黏膜层较致敏组变薄,水肿明显改善,炎性细胞明显降低,纤毛有所生长。IFN-γ组大鼠鼻黏膜层较致敏组变薄,水肿改善,仅见少量炎性细胞,纤毛有所生长。 结论 AR大鼠外周血中Th17、Treg细胞比例异常,伴有细胞因子IL-17A、TGF-β1表达水平的异常。IFN-γ可能通过下调AR大鼠外周血中Th17细胞比例及降低IL-17A水平,同时上调Treg细胞的比例及升高TGF-β1水平,对AR发挥治疗作用。

关键词: 动物模型, 变应性鼻炎, γ干扰素, Th17细胞, Treg细胞, 白介素17A, 转化生长因子, Wistar大鼠

Abstract: Objective To investigate the effects of interferon gamma(IFN-γ)on T helper cell 17(Th17)and regulatory T cell(Treg)(Th17/Treg)in peripheral blood of rats with allergic rhinitis(AR). Methods A total of 40 Wistar male rats were randomly divided into 4 groups: control group, sensitized group, triamcinolone acetonide group, and IFN-γ group. The control group was injected with saline intraperitoneally, and had ultra-pure water nasal drops. The other 3 groups were injected with ovalbumin(OVA)and aluminium hydroxide and had OVA nasal instillation to establish AR models. After that, the sensitized group was treated with 50 μL phosphate buffer(PBS), triamcinolone acetonide group was treated with 3.5 μg triamcinolone acetonide, and IFN-γ group was treated with 1 μg IFN-γ nasal drops on either nostril. The proportions of Th17 cells and Treg cells in peripheral blood were measured with flow cytometry within 24 hours after the last administration, and levels of interleukin-17 A(IL-17A)and transforming growth factor(TGF-β1)were detected with quantitative enzyme-linked immunosorbent assay(ELISA)within 1 week. The nasal mucosa was observed with hematoxylin-eosin(HE)staining. Results Compared with control group ［(0.213 4±0.149 5)%］, the percentage of Th17 cells in sensitized group ［(4.975 7±1.503 9)%］, triamcinolone acetonide group ［(1.169 3±0.386 1)%］ and IFN-γ group ［(1.532 0±0.526 5)%］ increased in varying degrees(F=51.532, P<0.05), with no statistical difference between the triamcinolone acetonide group and IFN-γ group(P>0.05), but with differences among the other groups(P<0.05). Compared with control group ［(22.939 3±2.099 5)%］, the proportion of Treg cells in sensitized group ［(8.223 2±2.1888)%］, triamcinolone acetonide group ［(14.087 5±1.369 6)%］ and IFN-γ group ［(16.614 3±1.621 7)%］ decreased in varying degrees(F=97.107, P<0.05), with no difference between the triamcinolone acetonide group and IFN-γ group(P>0.05), but with differences among the other groups(P<0.008 3). Compared with control group ［(5.076 9±1.435 6)pg/mL］, the concentration of IL-17A in sensitized group ［(34.900 5±11.670 8)pg/mL］, triamcinolone acetonide group ［(9.733 8±1.828 9)pg/mL］, and IFN-γ group ［(10.524 6±2.118 0)pg/mL］ increased(F=27.173, P<0.05), with no difference between the triamcinolone acetonide group and IFN-γ group(P>0.05), but with differences among the other groups(P<0.05). Compared with the control group ［(431.881 6±67.326 2)pg/mL］, the concentration of TGF-β1 in sensitization group ［(173.886 2±41.708 2)pg/mL］, triamcinolone acetonide group ［(246.053 3±13.841 2)pg/mL］ and IFN-γ group ［(290.012 7±22.885 6)pg/mL］ decreased(F=36.623, P<0.05), with no differences among the groups(P<0.05). Compared with control group, the nasal mucosa of sensitized group was thicker and more edematous, with infiltration of a large number of inflammatory cells, disordered cell arrangement, goblet cell proliferation and almost invisible cilia. The nasal mucosa of triamcinolone acetonide group was thinner, edema was improved, inflammatory cells were significantly reduced and cilia slightly grew. The nasal mucosa of IFN-γ group was thinner than that of sensitized group, edema was improved, only a few inflammatory cells were found and the cilia slightly grew. Conclusion The percentages of Th17 and Treg cells in peripheral blood of AR rats were abnormal, accompanied by abnormal expression of IL-17A and TGF-β1. IFN-γ may play a therapeutic role in AR by decreasing the proportion of Th17 cells and level of IL-17A, while increasing the proportion of Treg cells and level of TGF-β1.

Key words: Animal models, Allergic rhinitis, Interferon gamma, T helper cell 17, Treg cells, Interleukin-17A, Transforming growth factor, Wistar rats

中图分类号: 

  • R765
[1] Berker M, Frank LJ, Geβner AL, et al. Allergies-A T cells perspective in the era beyond the T1/T2 paradigm[J]. Clin Immunol(Orlando, Fla.), 2017, 174:73-83. doi:10.1016/j. clim.2016.11.001.
[2] Bajoriuniene I, Malakauskas K, Lavinskiene S, et al. Response of peripheral blood Th17 cells to inhaled Dermatophagoidespteronyssinus in patients with allergic rhinitis and asthma[J]. Lung, 2012, 190(5):487-495.
[3] Wang M, Gu Z, Yang J, et al. Changes among TGF-β1 Breg cells and helper T cell subsets in a murine model of allergic rhinitis with prolonged OVA challenge[J]. Int Immunopharmacol, 2019, 69:347-357.doi: 10.1016/j.intimp.2019.01.009.
[4] Zrioual S, Ecochard R, Tournadre A, et al. Genome-wide comparison between IL-17A and IL-17F-induced effects in human rheumatoid arthritis synoviocytes[J]. Immunol, 2009, 182(5):3112-3120.
[5] 李钦, 张永东, 孙崇伟, 等. 鼻腔吸入γ干扰素对大鼠变应性鼻炎的治疗作用[J]. 中华耳鼻咽喉头颈外科杂志, 2008, 43(2):134-138. LI Qin, ZHANG Yongdong, SUN Chongwei, et al. Effects of interferon gamma on Eosinophil apoptosis and Eotaxin levels in nasal lavage fluid of allergic rhinitis rats[J]. Chinese Journal of Otorhinolaryngology Head and Neck Surgery, 2008, 43(2):134-138.
[6] 李钦, 陈彦林, 马焱燚, 等. JNK信号途径蛋白P-JNK和P-c-Jun在变应性鼻炎大鼠鼻黏膜组织重塑中的作用[J]. 临床耳鼻咽喉头颈外科杂志, 2015, 29(23):2057-2059. LI Qin, CHEN Yanlin, MA Yanyan, et al. Role of P-JNK and P-c-Jun of JNK transduction pathway on the nasal mucosa remodeling in allergic rhinitis rats[J]. Journal of Clinical Otorhinolaryngology Head and Neck Surgery, 2015, 29(23):2057-2059.
[7] 张仲林, 钟玲, 袁明勇, 等. Th1/Th2/Th17/Treg细胞因子在变应性鼻炎发病中的免疫机制研究[J]. 医学综述, 2014, 20(16):2906-2909. ZHANG Zhonglin, ZHONG Ling, YUAN Mingyong, et al. Study on Th1/Th2/Th17/Treg in the Immune Mechanism of Allergic Rhinitis Onset[J]. Medical Recapitulate, 2014, 20(16):2906-2909.
[8] 张丽霞. Treg通过TICAM-1通路对变应性鼻炎发病机制的影响[J]. 临床耳鼻咽喉头颈外科杂志, 2018, 32(22): 1763-1766. ZHANG Lixia. Treg influences the pathogenesis of allergic rhinitis through TICAM-1 pathway[J]. Journal of Clinical Otorhinolaryngology Head and Neck Surgery, 2018, 32(22):1763-1766.
[9] Vroman H, Van Den Blink B, Kool M. Mode of dendritic cell activation: the decisive hand in Th2/Th17 cell differentiation.Implications in asthma severity[J]. Immunobiology, 2015, 220(2):254-261.
[10] Lubbert E. IL-17/Th17 targeting on the road to prevent chronicdestructive arthritis[J]. Cytokine, 2008, 41(2):84-91.
[11] Zhang H, Kong H, Zeng X, et al. Subsets of regulatory T cells and their roles in allergy[J]. J Transl Med, 2014, 12:125. doi:10.1186/1479-5876-12-125
[12] Robertsen B. The interferon system of teleost fish[J]. Fish Shellfish Immunol, 2006, 20(2):172-191.
[13] 肖伟, 余惠, 郑春燕, 等. 哮喘患者外周血单个核细胞中Th1/Th2 类细胞因子的偏移状态及红霉素的纠偏作用[J]. 中华结核和呼吸杂志, 2000, 23(6):347-350. XIAO Wei, YV Hui, ZHENG Chunyan, et al. The imbalance of TH1/TH2 cytokine expression in peripheral blood mononudear cell from asthmatic patients and the effect of erythromycin on these cytokines[J]. Chinese Journal of Tuberculosis and Respiratory Diseases, 2000, 23(6):347-350.
[14] Zhang J, Tang S. Establishment and evaluation of the SD rat allergic rhinitis model[J]. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za zhi, 2015, 29(15):1372-1374.
[15] Yang J, Zhong W, Xue K, et al. Epigenetic changes: An emerging potential pharmacological target in allergic rhinitis[J]. Int Immunopharmacol, 2019, 71:76-83. doi:10.1016/j.intimp.2019.03.004
[16] Li Y, Zhou J, Rui X, et al. PM2.5 exposure exacerbates allergic rhinitis in mice by increasing DNA methylation in the IFN-γ gene promoter in CD4+T cells via the ERK-DNMT pathway[J]. Toxicol Lett, 2019, 301:98-107. doi:10.1016/j.toxlet.2018.11.012.
[17] 李钦, 陈彦林, 马焱燚, 等. γ干扰素对变应性鼻炎大鼠鼻腔灌洗液中嗜酸粒细胞凋亡及Eotaxin水平的影响[J]. 山东大学耳鼻喉眼学报, 2016, 30(4):30-33. LI Qin, CHEN Yanlin, MA Yanyi, et al. Effects of interferon gamma on Eosinophil apoptosis and Eotaxin levels in nasal lavage fluid of allergic rhinitis rats[J]. J Otolaryngol Ophthalmol Shandong University, 2016, 30(4):30-33.
[18] 李钦, 张永东, 孙崇伟, 等. 鼻腔滴入IFN-γ对AR大鼠鼻腔黏膜中GATA-3表达的影响及意义[J]. 山东医药, 2009, 49(14):27-28. LI Qin, ZHANG Yongdong, SUN Chongwei et al. Treatment of allergic rhinitis rats by blockade of GATA3 expression with absorbed interferon gamma[J]. Shandong Medical Journal, 2009, 49(14):27-28.
[19] 张成, 洪苏玲, 胡国华. 变应性鼻炎患者Treg/Th17细胞相关转录因子及细胞因子的失衡表达[J]. 临床耳鼻咽喉头颈外科杂志, 2012, 26(5):209-211. ZHANG Cheng, HONG Sulin, HU Guohua. The expression of Treg/Th17 cells related transcription factors and cytokines in PBMCs and plasma in patients with allergic rhinitis [J]. Journal of Clinical Otorhinolaryngology Head and Neck Surgery, 2012, 26(5):209-211.
[20] Wang Y, Hou XR, Li LH, et al. Acupoint injection improves allergic rhinitis by balancing Th17/Treg in allergic rhinitis rats[J]. Zhen Ci Yan Jiu, 2019, 44(4):276-281.
[21] Jiao WE, Wei JF, Kong YG, et al. Notch signaling promotes development of allergic rhinitis by suppressing foxp3 expression and treg cell differentiation[J]. Int Arch Allergy Immunol, 2019, 178(1):33-44.
[22] Qiao Y, Liu T, Zhang KJ, et al. Allergic rhinitis management:environment control and patient education[J]. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za zhi, 2017, 31(17):1315-1321.
[23] Wen J, Zhou Y, Wang J, et al. Interactions between Th1 cells and Tregs affect regulation of hepatic fibrosis in biliary atresia through the IFN-γ/STAT1 pathway[J]. Cell Death Differ, 2017, 24(6):997-1006.
[24] 李钦, 陈彦林, 马焱燚, 等. γ干扰素抑制c-Jun氨基末端激酶磷酸化对大鼠变应性鼻炎鼻黏膜组织重塑的影响[J]. 临床耳鼻咽喉头颈外科杂志, 2016, 30(13):1034-1037. LI Qin, CHEN Yanlin, MA Yanyi, et al. Effects of inhibiting the phosphorylation of JNK by absorbed INF-γ on the remodeling of nasal mucosa in allergic rhinitis rats[J]. Journal of Clinical Otorhinolaryngology Head and Neck Surgery, 2016, 30(13):1034-1037.
[25] Jan L Bro(·overz)ek, Jean Bousquet, Carlos E, et al. Allergic Rhinitis and its Impact on Asthma(ARIA)guidelines: 2010 Revision[J]. J Allergy Clin Immunol, 2010, 126(3):466-476.
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