PPAR-γ激动剂对哮喘急性发作期患者T淋巴细胞磷酸化STAT6影响的体外实验研究  

Peroxisome proliferators-activated receptor gamma-agonist inhibits the expression of phosphorylation of signal transducer and activator of transcription-6 in T-lymphocytes of patients with acute asthma in vitro

作　　者：杨海华[1,2] 王文军 向旭东[1] 陈平[1] 黄玲[1,4] 邓军卫[1] 

机构地区：[1]中南大学湘雅二医院呼吸内科,湖南长沙410011 [2]上海徐汇区中心医院呼吸内科 [3]四川泸州医学院附属医院呼吸内科 [4]四川绵阳市三医院呼吸内科

出　　处：《中国呼吸与危重监护杂志》2008年第5期346-349,I0001,共5页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：湖南省科技厅资助项目(编号:05fj3035)

摘　　要：目的观察过氧化物酶增殖物激活受体γ(PPAR-γ)激动剂罗格列酮对哮喘急性发作期(简称哮喘急发)患者T淋巴细胞磷酸化信号转导和转录激活因子6(p-STAT6)及IL-4表达的影响,探讨罗格列酮的抗炎机制。方法选取健康对照组(A组)10例,哮喘急发患者10例,抽血、分离、纯化、培养外周血T淋巴细胞;其中,哮喘急发患者标本分为哮喘急发未干预组(B组,n=10)和哮喘急发罗格列酮干预组(C组,n=10),C组在T淋巴细胞培养开始时加入罗格列酮(浓度为10-4mol/L)。培养48 h后,采用ELISA法测定培养上清液IL-4浓度,Western blot法和免疫组织化学法检测T淋巴细胞p-STAT6的表达水平。结果B组患者的IL-4水平高于A组和C组[(170.34±9.05)pg/mL比(76.82±7.06)pg/mL和(123.59±8.70)pg/mL,P均<0.05],其中C组IL-4水平高于A组(P<0.05)。B组患者的p-STAT6表达水平高于A组和C组[Western blot法:6.28±0.19比3.07±0.18和4.12±0.16,P均<0.01;免疫组织化学法:(36.58±7.41)%比(11.39±4.02)%和(23.92±5.89)%,P均<0.05],其中C组p-STAT6表达水平高于A组(Western blot法:P<0.01;免疫组织化学法:P<0.05)。B组患者的IL-4水平与p-STAT6水平呈正相关(Western blot法:r=0.86,P<0.01;免疫组织化学法:r=0.82,P<0.01)。结论罗格列酮可抑制哮喘急发患者体外培养的T淋巴细胞p-STAT6的表达及IL-4的分泌。Objective To explore the possible anti-inflammatory mechanism of peroxisome proliferators-activated receptor（PPAR） gamma-agonists by investigating the effects of Rosiglitazone on the expression of phosphorylation of signal transducer and activator of transcription-6 （ p-STAT6 ） and the secretion of interleukin（IL）-4 in T-lymphocytes from patients with acute asthma. Methods Peripheral blood T-lymphocytes from 10 healthy volunteers （group A） and 10 patients with acute asthma were isolated, purifieated and cultured. T-lymphocytes from the asthma patients were divided into a control group （group B） and a Rosiglitazone treated group （group C ）. Rosiglitazone was added with a single dose of 10^ -4 mol/L at 0 hour of eultrue. After cultured for 48 hours, the concentration of IL-4 in supernatant of each group was detected by ELISA. The express of p-STAT6 in the T-lymphocytes was determined by Western blot and immunohistochemical techniques. Results The level of IL-4 was increased in group B than that in group A and group C [ （ 170. 34 ± 9.05 ） pg/mL vs （76. 82 ±7.06） pg/mL and （ 123.59 ± 8.70） pg/mL, both P 〈 0. 05 ] , and which in group C was higher than group A （P 〈 0.05 ）. The level of p-STAT6 in T lymphocyte in group B was higher than that in group A and C [ Western blot ：6. 28 ± 0. 19 vs 3.07±0.18 and 4. 12 ±0.16,all P 〈0.01;immunohistochemistry：（36.58 ±7.41）% vs （11.39 ± 4. 02）% and （23.92±5.89）%,all P 〈 0. 05 ], and which in group C was higher than that in group B [ Western hlot（ P 〈 0. 01 ）, immunohistochemistry （ P 〈 0. 05 ） 1. There was a positive correlation between the level of p-STAT6 and IL-4 （ P 〈 0. 01 ）. Conclusion The levels of p-STAT6 and IL-4 in T-lymphocytes of patients with acute asthma could be suppressed by Rosiglitazone in vitro.

关 键 词：哮喘 T淋巴细胞 过氧化物酶增殖物激活受体Γ 激动剂 罗格列酮 信号转导和转录激活因子6 

分 类 号：R562.25[医药卫生—呼吸系统]