18β-甘草次酸钠抑制变应性鼻炎大鼠鼻黏膜胸腺基质淋巴细胞生成素表达的研究  

The inhibition of 18p-sodium glycyrrhetinic acid on thymic stromal lymphopoietin expression in the nasal mucosa of allergic rhinitis rats

作　　者：季洁 桂岩[1] 王有虎[1] 侯赟[1] 陈康兵 席克虎[1] 陈小婉[1] 刘小涵 张小兵[1] Ji Jie;Gui Yan;Wang Youhu;Hou Yun;Chen Kangbing;Xi Kehu;Chen Xiaowan;Liu Xiaohan;Zhang Xiaobing(The First Clinical Medical College of Lanzhou University,Lanzhou 730030,China;Department of Otorhinolaryngology Head and Neck Surgeryythe First People's Hospital of Lanzhou, Lanzhou 730050,China)

机构地区：[1]兰州大学第一临床医学院,730030 [2]兰州市第一人民医院耳鼻咽喉头颈外科,730050

出　　处：《中华耳鼻咽喉头颈外科杂志》2019年第6期456-463,共8页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金地区科学基金项目(81160449).

摘　　要：目的探索18β-甘草次酸钠对变应性鼻炎(allergic rhinitis,AR)大鼠鼻黏膜胸腺基质淋巴细胞生成素(thymic stromal lymphopoietin ,TSLP)的作用。方法100只Wistar大鼠雌雄各半,采用随机数字表法随机分为5组,分别为对照组、AR模型组、布地奈德组、18β-甘草次酸钠20mg/kg剂量组和40mg/kg剂量组海组20只。除对照组外,其他4组以卵清蛋白(ovalbumin,OVA)致敏建立AR动物模型,造模成功后,布地奈德组、18β-甘草次酸钠20 mg/kg剂量组和40mg/kg剂量组分别给予布地奈德、18β-甘草次酸钠20 mg/kg和40mg/kg干预,检测时间点为用药2周和4周时。用免疫组织化学(免疫组化)法检测TSLP在大鼠鼻黏膜的分布情况;用免疫印迹法在蛋白水平检测大鼠鼻黏膜TSLP的表达;用实时荧光定量聚合酶链反应(RT-PCR)在基因水平检测大鼠鼻黏膜TSLP-mRNA的表达;用酶联免疫吸附测定法(EL1SA)检测大鼠血清中白细胞介素(interleukin,ID4和OVA特异性IgE(OVAspecific IgE,OVA-slgE)的浓度水平。多组间比较采用单因素方差分析和最小显著差异法,两组间比较采用LSD r检验,P<0.05为差异有统计学意义。结果免疫组化证实大鼠鼻黏膜中TSLP表达于上皮细胞、内皮细胞和上皮纤毛处。免疫印迹和KT-PCR提示AR模型组较对照组大鼠的鼻黏膜TSLP及 TSLP-mRNA 表达明显增强(2周TSLP:1.7959 + 0.1314比0.9905±0.1642,4周 TSLP:1.8097±0.2534比0.8703+0.1244;2周TSLP-mRNA: 4.5829±0.6977比1.1087±0.081 1,4 周 TSLP-mRNA:4.814 4±0.662 8比1.001 0±0.155 3;P值均<0.05)。给予药物干预2周、4周后,与AR模型组相比,布地奈德组、18β-甘草次酸钠20 mg/kg和40 mg/kg剂量组大鼠鼻黏膜TSLP及TSLP-mRNA的表达均被明显抑制[2 周 TSLP:(0.897 8±0.081 8)/(1.0721±0.1136)/(1.3966±0.1339)比1.7959±0.1314;4周TSLP:(1.1910±0.1613)/(1.1410±0.152 3)/( 1.200 5±0.189 6)比 1.809 7±0.253 4;2周TSLP-mRNA:(1.1756±0.1009)/(1.2544±0.078 2)/(2.0372±0.5592)比4.5829±0.697 7;4周TSLP-mRNA:(1.158Objective To explore the effect of 18β-soclium glycyrrhetinic acid on thymic stromal lymphopoietin (TSLP) in nasal mucosa of allergic rhinitis (AR) rats. Methods One hundred Wistar rats,half male and half female,were randomly divided into 5 groups by random number table method: control group, AR model group.budesonide group, 18β-sodium glycynhetinic acid at dose of 20 mg/kg and 40 mg/kg groups, with 20 rats in each group. AR animal models were established by ovalbumin (OVA) sensitization in the other four experimental groups. After successful modeling, budesonide and 18P-sodium glycyrrhetinic acid were given in each group,and the detection time points were 2 weeks and 4 weeks. The distribution of TSLP in rat nasal mucosa was detected by immunohistochemistry,and the expression of TSLP in rat nasal mucosa was determined l)y Western blot at the protein level. The expression of TSLP-mRNA in rat nasal mucosa was detected and compared by real-time fluorescence quantitative PCR (RT-PCR) at mRNA level. The concentrations of IL-4 and OVA-sIgE in rat serum were measured and compared by ELISA. One-way analysis of variance and the least significant difTerence method were used for the comparison among groups, LSD t test was used for the comparison between the two groups,and the difference was statistically significant (P<0.05). Results Immunohistochemistry confirmed existence of TSLP in rat nasal mucosa,especially in epithelial cells,endothelial cells and epithelial cilia. Western blot and RT-PCR suggested that the expression of TSLP and TSLP-mRNA in nasal mucosa of AR model group was significantly higher than that of control group (2 weeks TSLP: 1.7959±0.1314 % vs 0.9905±0.1642,4 weeks TSLP: 1.8097±0.2534 vs 0.8703± 0.1244;2 weeks TSLP-mRNA:4.5829±0.6977 vs 1.1087±0.0811.4 weeks TSLP-mRNA:4.8144±0.6628 vs 1.0010±0.1553;all P<0.05). After 2 weeks and 4 weeks of drug intervention,the expression of TSLP and TSLP-mRNA was inhibited in nasal mucosa of budesonide group. 18β-sodium sodium glycyrrhetinic acid at dose of 2

关 键 词：鼻炎 变应性 鼻黏膜 18β-甘草次酸钠 胸腺基质淋巴细胞生成素 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]