论文部分内容阅读
目的探讨吸入变应原引起支气管哮喘(简称哮喘)过敏性气道炎症免疫耐受形成的机制。方法BALB/c小鼠60只,按随机数字表法分为实验组(50只)和空白对照组(10只),实验组小鼠先给予腹腔注射卵清白蛋白(OVA)1mg,每周1次,共3周。雾化吸入OVA每天1h(含OVA80μg),连续10d。依据吸入OVA时间分为A、B、C、D、E5组,每组10只。A组雾化吸入10d后处死。B、D组继续每天1次,每次1h,每周5次,分别吸入OVA4周及8周,然后每天1h,连续10d吸入OVA后处死。C组停止吸入OVA4周后再次吸入OVA,每天1h,连续10d后处死。E组每天1次,每次1h,每周5次,吸入OVA4周,停止雾化吸入OVA4周,然后每天1h,连续10d吸入OVA后处死。测定各组小鼠支气管肺泡灌洗液(BALF)中细胞总数,嗜酸粒细胞、淋巴细胞、CD4+、CD8+、CD4+IL-10+分类及BALF中白细胞介素4(IL-4)、γ干扰素(IFN-γ)、IL-10的含量。测定血清中IL-4、IFN-γ、IL-10、OVA、IgE、IgG1、IgG2a水平,并对各组小鼠肺组织病理学进行分析。结果空白对照组BALF中嗜酸粒细胞、B淋巴细胞、CD4+IL-10+细胞分别为0.010±0.000、2.1±1.9、4.9±1.5,A组分别为0.480±0.110、5.1±2.6、5.1±2.3,B组分别为0.120±0.020、8.9±3.6、10.4±3.6,C组分别为0.560±0.050、4.7±1.7、6.3±3.1,D组分别为0.070±0.030、10.1±2.9、12.7±4.5,E组分别为0.680±0.030、5.6±3.2、6.1±3.4,各组间比较差异有统计学意义(F值分别为36.46、31.89、167.89,P均<0.01)。B、D组BALF中CD4+IL-10+细胞数与A组比较差异有统计学意义(q=5.8、6.4,P均<0.05);空白对照组BALF中IL-4、IL-10水平分别为(21±3)pg/ml、(44±12)pg/ml,A组分别为(128±23)pg/ml、(68±18)pg/ml,B组分别为(54±12)pg/ml、(127±27)pg/ml,C组分别为(133±21)pg/ml、(78±17)pg/ml,D组分别为(8±18)pg/ml、(135±34)pg/ml,E组分别为(143±26)pg/ml、(76±15)pg/ml,组间比较差异有统计学意义(F分别为37.20、143.78,P均<0.01)。B、D两组BALF中IL-10水平与A组比较差异有统计学意义(q分别为7.8、9.6,P均<0.05)。结论持续吸入变应原可使小鼠气道炎症减轻,产生免疫耐受,调节T淋巴细胞产生的IL-10参与了耐受形成。
Objective To investigate the mechanism of allergic airway inflammation immune tolerance induced by inhaled allergen in bronchial asthma (asthma). Methods Sixty BALB / c mice were randomly divided into experimental group (n = 50) and blank control group (n = 10). The mice in experimental group were given intraperitoneal injection of 1 mg ovalbumin (OVA) Times, a total of 3 weeks. Inhaled OVA daily inhalation 1h (containing OVA80μg) for 10d. According to inhaled OVA time divided into A, B, C, D, E5 group, 10 in each group. A group inhaled 10d after injection. Groups B and D continued once a day for 1 hour and 5 times a week respectively, and inhaled OVA for 4 weeks and 8 weeks respectively, followed by 1 hour daily for 10 days. C group inhaled OVA 4 weeks after inhalation of OVA again, 1h per day for 10d after the death. Group E was given once a day for 1 hour and 5 times a week. OVA was inhaled for 4 weeks. OVA was stopped by inhalation for 4 weeks, then inhaled for OVA for 10 days. The number of total cells, eosinophils, lymphocytes, CD4 +, CD8 +, CD4 + IL-10 + in bronchoalveolar lavage fluid (BALF) and the levels of interleukin-4, Interferon (IFN-γ), IL-10 content. The levels of IL-4, IFN-γ, IL-10, OVA, IgE, IgG1 and IgG2a in serum were measured and the lung histopathology of each group was analyzed. Results The numbers of eosinophils, B lymphocytes and CD4 + IL-10 + cells in BALF of control group were 0.010 ± 0.000, 2.1 ± 1.9 and 4.9 ± 1.5, respectively, while those in group A were 0.480 ± 0.110, 5.1 ± 2.6 and 5.1 ± 2.3, B group were 0.120 ± 0.020,8.9 ± 3.6,10.4 ± 3.6, C group were 0.560 ± 0.050,4.7 ± 1.7,6.3 ± 3.1, D group were 0.070 ± 0.030,10.1 ± 2.9,12.7 ± 4.5, E group were 0.680 ± 0.030,5.6 ± 3.2,6.1 ± 3.4, the difference between each group was statistically significant (F values were 36.46,31.89,167.89, P <0.01). The numbers of CD4 + IL-10 + cells in BALF in group B and group D were significantly different from those in group A (q = 5.8 and 6.4, P <0.05). The levels of IL-4 and IL-10 in BALF of group B (54 ± 12) pg / ml in group A were (128 ± 23) pg / ml and (68 ± 18) pg / ml in group A and (54 ± 12) pg / (133 ± 21) pg / ml and (78 ± 17) pg / ml respectively in group C, (8 ± 18) pg / ml and ± 34 pg / ml in group E and 143 ± 26 pg / ml in group E and 76 ± 15 pg / ml in group E respectively (F = 37.20 and 143.78, P <0.01, respectively) . The levels of IL-10 in BALF in B and D groups were significantly different from those in A group (q = 7.8 and 9.6, P <0.05 respectively). CONCLUSION: Continuous inhalation of allergen can relieve airway inflammation and induce immune tolerance in mice, and IL-10 regulated by T lymphocytes is involved in tolerance formation.