COPD大鼠对糖皮质激素不敏感与丝裂原活化蛋白激酶磷酸酶-1的关系  

The relationship between COPD rats insensitive to glucocorticoids and mitogen-activated protein kinase phosphatase-1

作　　者：吴洁[1] 辛晓峰[1] 戴伟[1] 张鹏鹏[1] 赵明[1] 方丽萍[1] 

机构地区：[1]南京大学医学院附属金陵医院(南京军区南京总医院)干部呼吸内科,南京医学硕士研究生210002

出　　处：《医学研究生学报》2016年第1期40-45,共6页Journal of Medical Postgraduates

摘　　要：目的慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)对激素不敏感与丝裂原活化蛋白激酶磷酸酶-1(mitogen-activated protein kinase phosphatase-1,MKP-1)密切相关。文中探讨COPD大鼠激素不敏感与肺组织中MKP-1的表达及p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38MAPK)信号通路的关系。方法将50只雄性SD大鼠按随机数字表法分为对照组、COPD空白组、P38MAPK抑制剂组、地塞米松组、P38MAPK抑制剂+地塞米松组,每组10只。采用LPS+烟熏4个月建立COPD模型。建模成功后P38MAPK抑制剂组、地塞米松组每天分别腹腔注射SB203580(100 mg/kg)、地塞米松(2 mg/kg),P38MAPK抑制剂+地塞米松组注射同等剂量的p38抑制剂和地塞米松,连续14 d。支气管肺泡灌洗液测定TNF-α和IL-8的浓度以及细胞计数;测定肺组织平均内衬间隔(MLI)和肺泡破坏指数(DI);取肺组织,用蛋白印迹分析法测MKP-1和P38MAPK的表达。结果 LPS+烟熏4个月大鼠肺顺应性、每分钟通气量均显著低于对照组(P<0.01);而气道阻力、MLI和DI则高于对照组。肺组织病理显示LPS+烟熏4个月大鼠肺气肿形成,模型建立。Western blot检测结果表明,COPD空白组MKP-1、P38MAPK的表达较对照组明显升高(P〈0.01),而较P38MAPK抑制剂+地塞米松组MKP-1降低[(100±11)%vs(131±22)%,P<0.05)];与COPD空白组比较,P38MAPK抑制剂+地塞米松组MKP-1升高[(100±11)%vs(131±22)%,P<0.05)];与COPD空白组P38MAPK表达[(201±76)%]比较,P38MAPK抑制剂组、P38MAPK抑制剂+地塞米松组[(32±3)%、(68±7)%]明显降低,差异有统计学意义(P<0.05);与P38MAPK抑制剂组比较,地塞米松组P38MAPK表达明显升高[(32±3)%vs(185±12)%,P<0.05];与地塞米松组比较,P38MAPK抑制剂+地塞米松组P38MAPK表达降低[(185±12)%vs(68±7)%,P<0.05]。肺泡灌洗液细胞数和TNF-α、IL-8浓度检测结果表明,COPD空白组与P38MAPK抑制剂组、P38MAPK抑制剂+地塞米松组比较,上述指标均升高(P<0.05)Objective Corticosteroid insensitivity in lung tissue of chronic obstructive pulmonary disease （ COPD ） is closely related of mitogen-activated protein kinase phosphatase-1 （ MKP-1 ）. To investigate the expression and activity of MKP-1, and the relation of p38 lightning mitogen-activated protein kinase （p38MAPK） signaling pathway and cortieosteroid insensitivity in lung tissue of COPD rats. Methods Fifty male Sprague-Dawley rats were randomly divided into control group, COPD group, P38MAPK inhibitor group, dexamethasone group and P38 MAPK inhibitors ＋ dexamethasone group （ n = 10 for each）. The rat model of COPD was estab- lished by cigarette smoking combined with intratracheal injection of lipopolysaccharide （LPS）. The P38MAPK inhibitor group were in- traperitoneally injected SB203580 （100 mg/kg） ; the dexamethasone group were intraperitoneally injected dexamethasone （2 mg/kg） ; the P38MAPK inhibitor ＋ dexamethasone group intraperitoneally injected the same dose of p38 inhibitor and dexamethasone, for 14 consecutive days. The bronchoalveolar lavage fluid levels of TNF- alpha, IL-8 and the change of cell count and the expression of MKP 1 and P38MAPK in pulmonary tissue were measured. Results The COPD group pulmonary compliance and minute ventilation were significantly lower than control group （ P 〈 0.01 ） ; but airway resistance, MLI and DI were significantly higher than control group. The pathology of lung tissue showed that emphysema was significant in the COPD group. Western blot test results show ed the expression of MKP-1 and P38MAPK were significantly increased in COPD group compared with controls group （ P 〈 0. O1 ） ; The expression of MPK-1 in lung tissue was significantly up regulated in P38MAPK inhibitor ＋ dexamethasone group compared with COPD group [ （100 ＋ 11 ）% vs （131±22）% ,P 〈0.05 ）] ; the expression of P38MAPK in lung tissue significantly up regulated in COPD group （210 ± 7.6） % compared with P38MAPK inhibitor group （ 32 ± 3 ）

关 键 词：慢性阻塞性肺疾病 地塞米松 丝裂原活化蛋白激酶-1 P38丝裂原活化蛋白激酶 

分 类 号：R563[医药卫生—呼吸系统]