Objective To explore the effect of 18β-sodium 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 glycyrrhetinic 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 18β-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 by 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 difference 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 20 mg/kg and 40 mg/kg group,which was significantly different from that of AR model group (2 weeks TSLP:(0.8978 ± 0.0818)/ (1.0721 ± 0.1136)/ (1.3966 ± 0.1339) vs 1.7959 ± 0.1314; 4 weeks TSLP:(1.1910 ± 0.1613)/ (1.1410 ± 0.1523)/ (1.2005 ± 0.1896) vs 1.8097 ± 0.2534; 2 weeks TSLP-mRNA:(1.1756 ± 0.1009)/ (1.2544 ± 0.0782)/ (2.0372 ± 0.5592) vs 4.5829 ± 0.6977; 4 weeks TSLP-mRNA:(1.1583 ± 0.1043)/ (1.2240 ± 0.0340)/ (1.2752 ± 0.0996) vs 4.8144 ± 0.6628; all P<0.05), and not significantly different from control group. With the inhibition of TSLP, the concentrations of IL-4 and OVA-sIgE in rat serum were also decreased. Conclusion 18β-sodium glycyrrhetinic acid has obvious inhibitory effect on TSLP in nasal mucosa of AR rats, which can control Th2 type immune inflammatory reaction.%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20 mg/kg剂量组和40 mg/kg剂量组,每组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40 mg/kg剂量组分别给予布地奈德、18β-甘草次酸钠20 mg/kg和40 mg/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(ELISA)检测大鼠血清中白细胞介素(interleukin,IL)4和OVA特异性IgE(OVA specific IgE,OVA-sIgE)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 t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RT-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±0.0811,4周TSLP-mRNA:4.8144±0.6628比1.0010±0.1553;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:(0.8978±0.0818)/(1.0721±0.1136)/(1.3966±0.1339)比1.7959±0.1314;4周TSLP:(1.1910±0.1613)/(1.1410±0.1523)/(1.2005±0.1896)比1.8097±0.2534;2周TSLP-mRNA:(1.1756±0.1009)/(1.2544±0.0782)/(2.0372±0.5592)比4.5829±0.6977;4周TSLP-mRNA:(1.1583±0.1043)/(1.2240±0.0340)/(1.2752±0.0996)比4.8144±0.6628;P值均<0.05],较对照组差异均无统计学意义.随着TSLP受抑制,血清IL-4、OVA-sIgE浓度也降低.结论 18β-甘草次酸钠对AR大鼠鼻黏膜TSLP有明显抑制作用,可以控制Th2型免疫炎性反应.
展开▼
