抑制趋化素样因子1对变应性鼻炎白细胞介素-9表达和嗜酸性粒细胞的影响  

Study on the effect of inhibiting chemokine-like factor 1 on the expression of interleukin-9 and eosinophils in allergic rhinitis

作　　者：张峻峥 时文杰 谢伟伟 李海亮[1] 樊宏[1] ZHANG Junzheng;SHI Wenjie;XIE Weiwei;LI Hailiang;FAN Hong(Department of Otorhinolaryngology,Haikou Hospital,Xiangya Medical College,Central South University,Haikou 570208,China)

机构地区：[1]中南大学湘雅医学院附属海口医院耳鼻咽喉科,海南海口570208

出　　处：《西部医学》2022年第8期1133-1139,共7页Medical Journal of West China

基　　金：海南省卫生健康行业科研项目(20A200190)。

摘　　要：目的探究抑制趋化素样因子1(CKLF1)对变应性鼻炎(AR)大鼠内白细胞介素-9(IL-9)与嗜酸性粒细胞的影响。方法40只SPF级SD大鼠随机分为对照组、模型组、CKLF1抗体组(AR+CKLF1-C19组)、丙酸氟替卡松组(AR+FP组),每组10只。除对照组外,其余各组大鼠均用卵清蛋白(OVA)和氢氧化铝[Al(OH)_(3)]致敏法诱导AR,AR+CKLF1-C19组AR激发前连续10 d分别在双侧鼻腔滴5μL CKLF1-C19肽干预,AR+FP组AR激发前连续10 d分别在双侧鼻腔滴5μL丙酸氟替卡松喷雾剂干预,对照组和模型组均同时在双侧鼻腔滴5μL生理盐水。末次鼻腔激发后,各组大鼠均进行表观行为学指标评分;ELISA法检测血清IL-9、免疫球蛋白E(IgE)、干扰素-γ(IFN-γ)水平;HE染色观察鼻黏膜组织嗜酸性粒细胞表达;瑞氏染色检测鼻腔内嗜酸性粒细胞数目;免疫组织化学染色检测鼻黏膜组织IL-9表达;实时荧光定量PCR和Western blot检测鼻黏膜组织嗜酸性粒细胞主要碱性蛋白(MBP)、嗜酸性粒细胞阳离子蛋白(ECP)、嗜酸性粒细胞过氧化物酶(EPO)在mRNA和蛋白水平上的表达。结果与对照组比较,模型组大鼠出现打喷嚏、流涕以及频繁抓鼻的现象,血清中IL-9、IgE水平升高,IFN-γ水平下降,鼻黏膜有水肿、黏膜层增厚以及黏膜上皮脱落的症状,嗜酸性粒细胞数目增加,鼻黏膜组织内IL-9阳性表达升高,MBP、ECP、EPO在mRNA和蛋白水平上的表达均上调(P<0.05);与模型组比较,AR+CKLF1-C19组和AR+FP组大鼠打喷嚏、流涕、抓鼻等行为减少,血清中IL-9、IgE水平下降,IFN-γ水平升高,鼻黏膜水肿及炎症细胞浸润减轻,嗜酸性粒细胞数目减少,鼻黏膜组织内IL-9阳性表达下降,同时,MBP、ECP、EPO在mRNA和蛋白水平上的表达也均下调(P<0.05)。结论抑制CKLF1能够明显降低变应性鼻炎大鼠嗜酸性粒细胞的浸润,抑制IL-9表达,从而对变应性鼻炎起到一定的治疗作用。Objective To explore the effect of inhibiting chemokine like factor-1(CKLF-1)on allergic rhinitis(AR)in rats with interleukin-9(IL-9)Influence with eosinophils.Methods 40 SD rats were randomly divided into control group(Control),model group(AR),CKLF1 antibody group(AR+CKLF1-C19),fluticasone propionate group(AR+FP),10 rats in each group.Except for the control group,the rats in the other groups were all induced AR with ovalbumin(OVA)and aluminum hydroxide[Al(OH)_(3)]sensitization method,during the modeling period,the AR+CKLF1-C19 group was instilled 5μL of CKLF1-C19 peptide in the bilateral nasal cavity for 10 consecutive days before AR challenge;in the AR+FP group,5μL fluticasone propionate spray were dripped into the bilateral nasal cavity for 10 consecutive days before AR challenge.Both the control group and the model group were instilled 5μL of saline in both nasal cavities at the same time.After the last nasal cavity challenge,rats in each group were scored with apparent behavior indicators.ELISA method was used to detect the level of serum IL-9,immunoglobulin E(IgE),interferon-γ(IFN-γ).HE staining was used to detect the expression of eosinophils in nasal mucosa tissue.Wright staining was used to detect the number of eosinophils in the nasal cavity.Immunohistochemical staining was used to detect the expression of IL-9 in nasal mucosa tissue.Real-time fluorescent quantitative PCR and Western blot was used to detect eosinophil major basic protein(MBP),eosinophil cationic protein(ECP),and eosinophil peroxidase in nasal mucosa tissue(Eosinophil peroxidase,EPO)expression at the mRNA and protein levels.Results Compared with the control group,rats in the model group had sneezing,runny nose and frequent snatching.The serum IL-9 and IgE levels increased,the IFN-γlevel decreased,and the nasal mucosa had edema and thickened mucosal layer.As well as the symptoms of mucosal epithelial shedding,the number of eosinophils increased,the positive expression of IL-9 in nasal mucosa tissue increased,and the expression of MB

关 键 词：变应性鼻炎 趋化素样因子1 嗜酸性粒细胞 白细胞介素-9 

分 类 号：R765.22[医药卫生—耳鼻咽喉科]