抗炎平喘药物对支气管哮喘小鼠骨髓CD+34造血细胞的影响

目的　观察糖皮质激素和白三烯受体修饰剂对支气管哮喘 (简称哮喘 )模型小鼠骨髓CD 3 4 造血细胞的影响 ,研究潜在的以骨髓为靶点的抗炎机制。方法　以 1%卵白蛋白 (OVA)致敏并激发BALB/c小鼠建立哮喘模型。在连续激发 2周期间分为 4组 ,每组 6只 ,分别给予生理盐水 (对照 ,A组 )、泼尼松 (B组 )、孟鲁斯特 (C组 )及泼尼松 孟鲁斯特 (D组 )灌胃 ,末次激发后 2 4h分别取支气管肺泡灌洗液 (BALF)、外周血及骨髓 ,测定BALF、外周血中有核细胞的分类计数及骨髓中有核细胞总数 ;用流式细胞仪测定外周血及骨髓中CD 3 4 造血细胞、CD 4、CD 8T淋巴细胞占有核细胞的比例 ;免疫组化结合原位杂交法检测骨髓内表达白细胞介素 5受体α链 (IL 5Rα)mRNA的CD 3 4 造血细胞 (CD 3 4 IL 5RαmRNA 细胞 )计数。结果　A组BALF中嗜酸粒细胞 (EOS)计数为 ( 18 3± 1 3)×10 5/L]、外周血中EOS为 ( 2 5± 0 4 )× 10 8/L]、CD 3 4 造血细胞为 ( 9 6± 5 1)× 10 7/L]、骨髓中CD 3 4造血细胞为 ( 7 7± 3 2 )× 10 7/根股骨 ];B组分别为 ( 4 6± 1 7)× 10 5/L]、( 1 5± 0 3)× 10 8/L]、( 3 9± 2 1)× 10 7/L]、( 3 3± 1 8)× 10 7/根股骨 ];D组分别为 ( 3 7± 1 4 )× 10 5/L]、( 1 7± 0 3)× 10 8/L]

The effects of anti-inflammatory and anti-asthmatic agents on CD+34 hematopoietic cells in bone marrow of asthmatic mice

OBJECTIVE: To observe the effects of glucocorticoids and cysteinyl leukotrienes 1 receptor antagonist on CD(34)(+) hematopoietic cells, and to study the rationality of a bone marrow-targeting anti-inflammatory strategy. METHODS: Twenty-four BALB/c mice were sensitized and challenged by 1% ovalbumin (OVA) to establish the asthmatic model. Asthmatic mice were challenged by 1% OVA and divided into 4 groups: fed by sterile saline (group A), prednisone (group B), montelukast (group C) and prednisone plus montelukast (group D) respectively for two consecutive weeks. The mice were killed at 24 h after the last challenge, then bronchoalveolar lavage fluid (BALF), peripheral blood and bone marrow were prepared. Eosinophils in peripheral blood and BALF, nucleate cells in BALF, peripheral blood and bone marrow were counted. The percentage of CD(34)(+) cells, CD(4)(+), CD(8)(+) T lymphocyte to nucleate cells in peripheral blood and bone marrow were counted by flow cytometry. Immunocytochemistry and in situ hybridization were employed to detect the hematopoietic cells expression of CD(34)(+) and IL-5Ralpha mRNA in bone marrow (CD(34)(+) IL-5Ralpha mRNA(+) cells). RESULTS: The number of EOS in BALF and peripheral blood and the number of CD(34)(+) cells in peripheral blood and bone marrow in group A were (18.3 +/- 1.3) x 10(5)/L], (2.5 +/- 0.4) x 10(8)/L], (9.6 +/- 5.1) x 10(7)/L] and (7.7 +/- 3.2) x 10(7)/femur] respectively, compared with the corresponding indices in group B (4.6 +/- 1.7) x 10(5)/L, (1.5 +/- 0.3) x 10(8)/L, (3.9 +/- 2.1) x 10(7)/L, (3.3 +/- 1.8) x 10(7)/femur] and group D (3.7 +/- 1.4) x 10(5)/L, (1.7 +/- 0.3) x 10(8)/L, (4.1 +/- 1.8) x 10(7)/L, (2.2 +/- 0.7) x 10(7)/femur]; the differences all were significant (all P < 0.01). The number of bone marrow CD(34)(+) IL-5Ralpha mRNA(+) in group B and D were (23 +/- 7)% and (21 +/- 4)%, as compared with the corresponding index in group A (37 +/- 4)%], the differences were significant (P < 0.01); the number of eosinophils in BALF in group C was (12.2 +/- 1.1) x 10(5)/L, as compared with the corresponding index in group A (18.3 +/- 1.3) x 10(5)/L], the difference was significant (P < 0.05). CONCLUSIONS: Prednisone probably inhibits the proliferation, differentiation and emigration of CD(34)(+) cells in the bone marrow of asthmatic mice, and inhibits eosinophilopoiesis in bone marrow, eosinophil migration into peripheral blood and recruitment to the airways. Montelukast may suppress eosinophil infiltrating into lungs of asthmatic mice, but it does not inhibit the proliferation and emigration of CD(34)(+) cells and does not show apparent synergistic effect with prednisone.