细胞周期蛋白D_1在支气管哮喘小鼠肺组织中的表达及其与气道重塑的关系  

Cyclin D_1 and its association with airway remodeling in a murine model of asthma

作　　者：葛海燕[1] 黄茂[2] 姚欣[2] 殷凯生[2] 杨玉[2] 

机构地区：[1]南通大学附属医院呼吸科 [2]南京医科大学第一附属医院呼吸科,210029

出　　处：《中华结核和呼吸杂志》2006年第9期607-611,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：南京医科大学科技创新基金(CX2004011)

摘　　要：目的研究细胞周期蛋白 D_1(Cyclin D_1)在支气管哮喘(简称哮喘)小鼠肺组织中的表达,探讨 Cyclin D_1在哮喘及气道重塑中的作用。方法将40只 SPF 级 BALB/c 小鼠按随机数字表法分为正常对照组(A 组)、哮喘雾化2周组(B 组)、哮喘雾化4周组(C 组)、哮喘雾化8周组(D 组)4组,每组10只。用10%鸡卵白蛋白(OVA)致敏和1%OVA 激发小鼠建立哮喘模型。分析支气管肺泡灌洗液(BALF)中嗜酸粒细胞(EOS)计数及分类;用动物肺功能仪检测各组小鼠肺功能状况;用苏木精-伊红(HE)染色观察气道炎症及细胞浸润情况;用图像分析软件观察气道壁及平滑肌层变化情况;用逆转录-聚合酶链反应(RT-PCR)及实时定量(Real-time)PCR 测定肺组织中 Cyclin D_1mRNA 水平表达变化;用 Western blot 法观察肺组织中 Cyclin D_1的蛋白表达变化。结果 BALF 分析结果提示,B、C、D 组 EOS 计数分别为(42.6±0.9)×10~4/L、(54.7±1.4)×10~4/L、(44.8±2.4)×10~4/L,与A组[(3.4±0.5)×10~4/L]比较差异有统计学意义(q 值分别为79.75、91.42、84.82,P 均<0.01);对小鼠呼气阻力检测发现,乙酰胆碱浓度为45 μg/kg 时 B、C、D 组分别为(5.27±0.16)cm·L^(-1)·min^(-1)、(6.68±0.20)cm·L^(-1)·min^(-1)、(7.14±0.41)cm·L^(-1)·min^(-1),与 A 组[(4.11±0.15)cm·L^(-1)·min^(-1)]比较差异有统计学意义(q 值分别为5.58、6.39、7.11.P 均<0.05);支气管平滑肌面积/管腔内周长(WAm/Pi)B 组为2.8±0.6,C 组为4.8±0.6,D 组为6.4±0.7,与 A 组(2.4±0.4)比较差异有统计学意义(q 值分别为6.40、8.28、9.27,P<0.05);管壁面积/管腔内周长(WAt/Pi)B 组为6.4±0.8,C 组为8.3±1.2,D 组为9.3±1.0,与 A 组(5.6±1.0)比较差异有统计学意义(q 值分别为2.80、4.83、6.37,P 均<0.05);Western blot 检测发现 Cyclin D_1在 B、C、D 组表达量分别为0.587±0.015、0.808±0.029、0.826±0.022,与 A 组(0.404±0.016)比较差异有统计学意义(q 值分别为5.87、8.08、8.26,P 均<0.01);相关性分析结果提示呼�Objective To evaluate the expression of cyclin D1 in asthmatic mouse lungs, and to explore the role of cyclin D1 in bronchial asthma and airway remodeling. Methods Forty BALB/c mice were randomized to group A （ normal）, group B （ sensitized for 2 weeks）, group C （ sensitized for 4 weeks） and group D（sensitized for 8 weeks）, 10 mice each group. The mice were sensitized with 10% ovalbumin and challenged with 1% ovalbumin to establish the asthmatic model. The number of eosinophils and the cell percentages in bronchoalveolar lavage fluid （BALF） were counted by cytology method. Pulmonary functions were measured to evaluate the resistance of expiration. Airway inflammation and eosinophil infiltration were evaluated by HE staining, and the airway wall thickness （WAt/Pi） , smooth muscle thickness （WAm/Pi） and smooth nucleus counts（N/Pi） were quantified by computer-assisted image analysis system. The mRNA expression of cyclin D~ was measured by RT-PCR and Real-time PCR. The protein expression of cyclin D1 was assayed by Western blot. The correlation between airway resistance of expiration and the expression of cyclin D1 was studied. Results The eosinophil count and differential in BALF of group B, C, and D [ （42.6 ±0. 9） × 10^4/L, （54. 7 ± 1.4） × 10^4/L, （44. 8 ±2.4） × 10^4/L ] were higher than those of group A （3.4 ± 0. 5） × 10^4/L （ q = 79. 75,91.42,84. 82, all P 〈 0. 01 ）. The airway resistance of expiration after challenge with 45 μg/kg acetylcholine in group B,C,and D [ （5. 27 ±0. 16）cm · L^-1 · min^-1, （6.68 ± 0.20）cm·L^-1 · min^-1, （7. 14±0.41）cm· L^-1· min^-1] was higher than that in group A [（4.11 ± 0. 15） cm · L^-1 ·min^-1, q = 5.58 ,6. 39,7. 11 , all P 〈 0. 05 ] . Eosinophil infiltration, cilium loss, formation of mucus plug and smooth muscle cell layer thickening were observed in group B, C, and D. The morphological changes of the airways became evident following airway remodeling. WAm/Pi in group B, C, and D（2. 8 ±

关 键 词：细胞周期蛋白D1 哮喘 支气管高反应性 气道重塑 

分 类 号：R562.25[医药卫生—呼吸系统]