孟鲁司特对变应性鼻炎大鼠Th1/Th2失衡的免疫调节作用研究

Study of immunomodulatory effects of montelukast on the Th1/Th2 imbalance in rats with allergic rhinitis

目的研究孟鲁司特对变应性鼻炎(AR)大鼠辅助T淋巴细胞1(Th1)/辅助T淋巴细胞2(Th2)失衡的免疫调节作用。方法将SD大鼠88只随机分为空白组(n=22)、模型组(n=22)、实验组(n=22)及对照组(n=22)。模型组、实验组及对照组大鼠均用卵清蛋白(OVA)致敏法建立AR大鼠模型。造模成功后,实验组大鼠给予孟鲁司特(15 mg·kg^-1·d^-1)灌胃干预,对照组大鼠给予地塞米松(1 mg·kg^-1·d^-1)灌胃干预,空白组与模型组灌胃等量生理盐水,连续干预10 d。用鼻部症状行为学评分法记录各组大鼠AR症状行为;用酶联免疫吸附(ELISA)法检测血清OVA特异性免疫球蛋白E(OVA-sIgE)、白细胞介素-4(IL-4)及干扰素-γ(INF-γ)水平;用苏木精-伊红(HE)染色观察大鼠鼻黏膜组织病理变化;用免疫组化法检测鼻黏膜组织细胞间黏附分子-1(ICAM-1)、转化生长因子-β1(TGF-β1)蛋白表达。结果空白组、模型组、实验组及对照组大鼠症状行为评分分别为(0.76±0.35),(6.46±0.71),(3.42±0.59),(2.83±0.38)分;血清OVA-sIgE水平分别为(158.32±24.64),(973.12±68.25),(314.37±32.42),(226.58±28.16)ng·L^-1;IL-4水平分别为(10.87±0.96),(68.22±3.47),(27.83±1.58),(16.56±1.42)pg·mL^-1;INF-γ水平分别为(71.25±3.36),(21.68±3.11),(52.72±3.61),(64.33±2.84)pg·mL^-1;各组大鼠鼻黏膜组织ICAM-1蛋白阳性细胞百分比分别为(11.02±2.23)%,(86.34±3.76)%,(58.22±2.65)%,(21.32±1.93)%;TGF-β1蛋白阳性细胞百分比分别为(14.56±1.87)%,(72.08±2.83)%,(41.12±3.11)%,(20.42±2.04)%。与空白组相比,模型组大鼠症状行为评分,血清OVA-sIgE、IL-4水平及鼻黏膜组织ICAM-1与TGF-β1蛋白阳性细胞百分比均显著升高,且实验组低于模型组,对照组低于实验组;血清INF-γ水平降低,且实验组高于模型组,对照组高于实验组(均P<0.05)。结论孟鲁司特可降低AR大鼠血清OVA-sIgE水平,调节Thl/Th2细胞因子平衡,同时降低鼻黏膜组织中ICAM-1与TGF-β1蛋白表达,进而达到治疗AR的目的。

Objective To investigate the immunomodulatory effects of montelukast on the helper T lymphocyte 1(Th1)/helper T lymphocyte 2(Th2)imbalance in rats with allergic rhinitis(AR).Methods Eighty-eight SD rats were randomly divided into blank group(n=22),model group(n=22),test group(n=22)and control group(n=22).The AR rat models were established by ovalbumin(OVA)sensitization method in model group,test group and control group.After successful modeling,rats in test group were given montelukast(15 mg·kg^-1·d^-1)for intragastric intervention,rats in control group were given dexamethasone(1 mg·kg^-1·d^-1)for intragastric intervention,blank group and model group were given the same amount of normal saline for 10 d.The symptomatic behavior of AR in each group was recorded by nasal symptom behavioral scoring method.Serum OVA-specific immunoglobulin E(OVA-s IgE),interleukin-4(IL-4)and interferon-γ(INF-γ)levels were detected by enzyme-linked immunosorbent assay(ELISA).The pathological changes of rat nasal mucosa were observed by hematoxylin-eosin(HE)staining.The expression of intercellular adhesion molecule-1(ICAM-1)and transforming growth factor-β1(TGF-β1)protein in nasal mucosa were detected by immunohistochemistry method.Results The symptom behavior scores of blank group,model group,test group and control group were(0.76±0.35),(6.46±0.71),(3.42±0.59)and(2.83±0.38)scores,the serum OVA-s IgE levels were(158.32±24.64),(973.12±68.25),(314.37±32.42),(226.58±28.16)ng·L^-1,IL-4 levels were(10.87±0.96),(68.22±3.47),(27.83±1.58),(16.56±1.42)pg·m L^-1,the INF-γlevels were(71.25±3.36),(21.68±3.11),(52.72±3.61),(64.33±2.84)pg·m L^-1,the percentages of ICAM-1 protein positive cells in nasal mucosa of each group rats were(11.02±2.23)%,(86.34±3.76)%,(58.22±2.65)%,(21.32±1.93)%,the percentage of TGF-β1 protein positive cells were(14.56±1.87)%,(72.08±2.83)%,(41.12±3.11)%and(20.42±2.04)%.Compared with blank group,the symptomatic behavior scores,serum OVA-s IgE,IL-4 levels and the percentage of ICAM-1 and TGF-β1 protein positive cells in the nasal mucosa significantly increased in model group,test group were lower than model group,and control group were lower than test group;serum INF-γlevels significantly decreased,and test group was higher than model group,and control group was higher than test group(all P<0.05).Conclusion Montelukast can reduce the level of serum OVA-s IgE in AR rats,regulate the balance of Th1/Th2 cytokines,and decrease the expression of ICAM-1 and TGF-β1 protein in nasal mucosa,so as to achieve the purpose of treating AR.