支气管哮喘豚鼠肺组织中丝裂原活化蛋白激酶对γ-谷氨酰半胱氨酸合酶的作用  

Effects of mitogen activated protein kinase on γ-glutamylcysteine synthase in lung of guinea pigs with bronchial asthma

作　　者：朱运福 戴爱国 胡瑞成 

机构地区：[1]湖南省老年医院湖南省老年医学研究所呼吸疾病研究室,湖南长沙410001

出　　处：《中国病理生理杂志》2007年第10期1990-1995,共6页Chinese Journal of Pathophysiology

基　　金：湖南省科技计划资助项目(No.05JT1024);湖南省教育厅科研资助项目(No.02C408)

摘　　要：目的:研究在支气管哮喘豚鼠肺组织中丝裂原活化蛋白激酶(MAPK)信号通路对γ-谷氨酰半胱氨酸合酶(γ-GCS)的影响。方法:成年雄性豚鼠20只随机分成哮喘组和对照组,每组10只,以腹腔内注射联合雾化吸入卵蛋白复制哮喘模型。测支气管肺泡灌洗液(BALF)细胞总数并进行分类计数;原位杂交和RT-PCR测肺组织中γ-GCS-h mRNA表达;免疫组化测γ-GCS、磷酸化细胞外激活蛋白激酶(p-ERK)、磷酸化c-Jun氨基末端激酶(p-JNK)和磷酸化p38(p-p38)在肺组织中的表达;W estern b lotting测p-ERK、p-JNK和p-p38在肺组织中的表达;双酶法测γ-GCS的活性。结果:(1)哮喘组BALF中细胞总数、嗜酸性粒细胞明显多于对照组(P<0.01)。(2)免疫组化显示肺组织中p-ERK、p-p38、p-JNK和γ-GCS蛋白质表达哮喘组明显高于对照组(P<0.01);W estern b lotting亦显示肺组织中p-ERK、p-JNK和p-p38蛋白质表达哮喘组均明显高于对照组。(3)原位杂交和RT-PCR显示在肺组织中γ-GCS-h mRNA表达哮喘组明显高于对照组(P<0.01)。(4)γ-GCS活性哮喘组明显高于对照组(P<0.01)。(5)直线相关性分析显示:在肺组织中p-ERK、p-p38与γ-GCS-hmRNA、γ-GCS蛋白质呈显著正相关,p-JNK与γ-GCS-h mRNA、γ-GCS蛋白质无明显相关性。结论:支气管哮喘豚鼠肺中p-ERK、p-p38、p-JNK和γ-GCS表达均增加;p-ERK和p-p38可能上调γ-GCS表达。AIM ： To investigate the effects of mitogen activated protein kinase on γ - glutamylcysteine synthase （ γ - GCS） in lung of guinea pigs with bronchial asthma. METHODS ： Twenty adult male guinea pigs were divided into asthmatic group and control group （ 10 in each group）. Asthmatic model was established by ovalbumin intraperitoneal injection combined with inhalation, The numbers of total and inflammation cells in bronchoalveolar lavage fluid （BALF） were measured. The γ - GCS - h mRNA in lung tissue was examined by in situ hybridization and RT - PCR, Immunohistochemistry was used to detecte the expression of γ- GCS, phosphorylated extracellular signal regulated kinase （p- ERK） , phosphrylated c -Jun amino terminal kinase （p -JNK） and phosphorylated p38 （p -p38 ） in lung tissues. Western blotting was conducted to determine the expressions of p - ERK, p - JNK and p - p38 in lung tissue. The activity of γ - GCS was measured by coupled enzyme assay. RESULTS： （ 1 ） The total cell number and number of eosinophils in BALF of asthmatic group were significantly higher than those in control group （ P 〈 0.01 ）, （2） Immunohistochemistry indicated that the p-ERK, p- p38, p- JNK and γ- GCS were stronger expressed in asthmatic group than those in control group （ P 〈0. 01 ）. Western blotting also discovered that the expressions of p - ERK, p - JNK and p - p38 in lung tissue of asthmatic group were stronger than those in control group. （ 3 ） Both in situ hybridization and RT - PCR analysis showed that the expression of γ- GCS- h mRNA was more positive in asthmatic group compared with control group （P 〈0. 01 ）. （4） The activity of γ- GCS of asthmatic grouo was significantly higher than that in control group （ P 〈0. 01 ）. （ 5 ） Linear cor-relation analysis indicated that in lung tissue of guinea pig with asthma, p - ERK and p - p38 markedly positive correlated with γ - GCS - h mRAN and γ- GCS protein. No relationship between p - JNK andγ - GCS - h mR

关 键 词：哮喘 有丝分裂素激活蛋白激酶类 谷氨酰半胱氨酸合酶 

分 类 号：R562[医药卫生—呼吸系统]