速激肽-1受体小干扰RNA对变应性鼻炎多种炎性因子表达的影响  

The effect of NK-1R-siRNA on expression of inflammatory factors in allergic rhinitis

作　　者：王虹 吴静 章如新 Wang Hong;Wu Jing;Zhang Ruxin(Department of Otolaryngology,Huadong Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华东医院耳鼻咽喉科,上海200040

出　　处：《中华耳鼻咽喉头颈外科杂志》2019年第4期280-285,共6页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金(30872848).

摘　　要：目的探讨速激肽-1受体小干扰RNA(neurokinin-1 receptor small interfering RNA,NK-1R-siRNA)对变应性鼻炎(allergic rhinitis,AR)鼻黏膜多种炎性因子表达的影响。方法将24只健康雄性SD大鼠采用随机数字表法随机分为3组,分别为NK-1R-siRNA组、阴性小干扰RNA (negative control-small interfering RNA,NC-siRNA)组和生理盐水组,每组8只。将所有大鼠用卵清蛋白(ovalbumin,OVA)建立AR动物模型,同时用NK-1R-siRNA、NC-siRNA和生理盐水滴鼻干预。观察各组大鼠的AR症状。酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测血清中OVA特异性IgE。实时定量聚合酶链反应(real time PCR,RT-PCR)及免疫组织化学(免疫组化)法检测各组大鼠鼻黏膜中NK-1R的表达情况。通过蛋白芯片技术比较各组大鼠鼻黏膜局部炎性因子的表达。使用SPSS 11.0软件,以单因素方差分析进行数据分析。结果与生理盐水组比较,NK-1R-siRNA组大鼠AR症状明显缓解[挠鼻(31.4±8.9)次/15 min比(69.5±17.9)次/15 min,喷嚏(7.2±1.9)次/15 min比(23.7±9.2)次/15 min,鼻分泌物(7.1±2.3)mg比(24.1±4.4)mg,t值分别为38.100、17.125、16.837,P值均<0.01]。与生理盐水组比较,NK-1R-siRNA组大鼠血清中OVA特异性IgE水平明显降低[(8.56±0.73)ng/ml比(18.05±1.22)ng/ml,t=9.787,P<0.01]。RT-PCR和免疫组化显示NK-1R-siRNA组大鼠鼻黏膜中NK-1R表达较NC-siRNA组和生理盐水组显著下降。蛋白芯片结果显示NK-1R-siRNA干预后,AR大鼠鼻黏膜中白细胞介素(interleukin,IL)1α、IL-1β、IL-4、IL-6、IL-13等多种炎性因子表达受到抑制,而γ干扰素(interferon-γ,IFN-γ)、IL-10的表达明显升高。结论 NK-1R-siRNA可通过调节多种AR相关炎性因子的表达,减轻鼻黏膜局部变应性炎症。Objective To explore the effect of neurokinin-1 receptor small interfering RNA (NK-1R-siRNA) on the expression of inflammation factors in allergic rhinitis (AR). Methods Twenty-four male SD rats were divided into three groups randomly (by random number table methord): NK-1R-siRNA group, negative control siRNA (NC-siRNA) group and saline group, with 8 rats in each group. SD rats were sensitized and challenged with ovalbumin (OVA) to induce AR. The rats were treated intranasally with NK-1R-siRNA, NC-siRNA or saline before and during the challenge period. The AR symptoms were observed. The levels of OVA-specific IgE were measured by enzyme-linked immunosorbent assay (ELISA). The levels of NK-1R expression in the nasal mucosal tissues were determined by real time PCR (RT-PCR) and immunohistochemistry. Antibody array was used in studying the expression of inflammation cell factors in nasal mucosa. SPSS 11.0 software was used for one-factor analysis of variance. Results Compared with saline group, AR symptoms relived significantly in NK-1R-siRNA group (nose rubbing (31.4±8.9)/15 min vs (69.5±17.9)/15 min, sneezing (7.2±1.9)/15 min vs (23.7±9.2)/15 min, nasal secretions (7.1±2.3) mg vs (24.1±4.4) mg, t value was 38.100, 17.125, 16.837, respectively, all P<0.01), and the level of serum OVA-specific IgE was also reduced ((8.56±0.73) ng/ml vs (18.05±1.22) ng/ml, t=9.787, P<0.01). The RT-PCR and immunohistochemistry results showed that the expression of NK-1R in nasal mucosa of NK-1R-siRNA group was remarkably reduced than that of the NC-siRNA group and saline group. After the treatment of NK-1R-siRNA, the expression of interleukin (IL) 1α, IL-1β, IL-4, IL-6 and IL-13 decreased, while the interferon-γ(IFN-γ) and IL-10 increased. Conclusion NK-1R-siRNA could regulate the release of inflammation factors in AR nasal mucosa, thus relive the allergic inflammation.

关 键 词：速激肽-1受体小干扰RNA 鼻炎 变应性 炎性因子 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]