转录因子GATA-3和T—bet的表达失衡在复苏后急性肺损伤中的作用  

Role of imbalance between transcription factors GATA-3 and T-bet expressions in the pathogenesis ofacute lung injury after resuscitation

作　　者：顾伟[1] 李春盛[1] 殷文朋[1] 张健[1] 侯晓敏[1] 张达[1] 

机构地区：[1]首都医科大学北京朝阳医院急诊科,北京100020

出　　处：《中华急诊医学杂志》2014年第1期8-14,共7页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金（30972863）;2012北京市优秀博士学位论文专项资金项目（2012-1002501）

摘　　要：目的探讨转录因子GATA-3和T．bet的表达失衡是否参与了猪心肺复苏后的急性肺损伤机制。方法26头猪随机（随机数字法）分为假手术组（SHAM）（6只）、复苏12h组和复苏24h组（各10只）。建立室颤8min小型猪模型；用流式细胞术检测猪外周血淋巴细胞（PBL）中CD3＋、CD4＋和CD8＋淋巴细胞亚群的分布；采用酶联免疫吸附（ELISA）方法检测血浆中白细胞介素4（IL4），肿瘤坏死因子仅（TNF—d）和干扰素1（INF-1）的含量；采用Western-blot法检测肺组织GATA．3和T—bet的蛋白表达；用实时荧光PCR法定量肺组织GATA一3和T—betmRNA水平。结果复苏组小猪发生严重肺组织细胞损伤，在复苏后的12h和24h，猪血清中CD4＋细胞的比例（28．4±2．3）％，（24．1±1．6）％明显低于假手术组（48．4±2．9）％，（51．1±5．4）％；CD4＋／CD8＋（1．7±0．9），（1．5±1．0）也较假手术组明显减低（2．5±1．3），（2．7±1．1）（P〈0．05）；IL-4和TNF-d水平在2～12h逐渐升高（P〈0．01）；INF-1水平和INF-y／IL4却在2～12h逐渐降低。复苏组的肺组织GATA-3的蛋白表达较假手术组分别在12h、24h明显升高，T—bet的蛋白表达和T—bet/GATA-3则分别在12h、24h明显降低。复苏组的肺组织GATAd和T—bet的mRNA水平较假手术组分别在12h、24h明显升高和减低。结论转录因子GATA-3和T-bet的表达失衡导致的肺免疫功能障碍可能参与了心肺复苏后急性肺组织损伤的过程。Objective To study the role of imbalance between transcription factors GATA-3 and T-bet expressions in causing acute lung injury after resuscitation in cardiac arrest model of swine. Methods After swine model of electrically induced cardiac arrest was established for 8 minutes, animals were resuscitated to get restoration of spontaneous circulation （ ROSC ）. The swine with ROSC were randomly assigned to be sacrificed at 12 and 24 h after ROSC （n =8 in each group）. CD3＋ , CIM＋ and CD8＋ lymphocyte subsets were determined by flow cytometry, and the levels of serum IL-4, TNF-a, and IFN-ywere measured by using ELLSA. The protein levels and expressions of GATA-3/T-bet mRNA were detected in lung tissue by western blotting and quantitative real-time PCR device, respectively. Results Pulmonary function was significantly impaired after ROSC. CIM ＋ lymphocyte subsets （28.4 ± 2. 3 ） %, （ 24. 1 ± 1.6） % and CIM ＋/CD8 ＋ （ 1.7 ± 0. 9）, （ 1.5 ±1.0） were significantly lower in the post-ROSC group compared with the sham-operated group （48.4＋_2.9）%, （51.1＋5.4）% （2.5±1.3）, （2.7±1.1） （P〈0.05） at12 h and 24 h after ROSC. The levels of serum IL-4 and TNF-a were markedly increased, while IFN-y and IFN-y/IL-4 were significantly decreased in the post-ROSC group compared with the sham-operated group （ P 〈 0. 05） at 2-12 hafter ROSC. Protein level and expression of GATA-3 mRNA in lung tissue were markedly increased, while those of T-bet were significantly reduced in the post-ROSC group compared with the sham-operated group （P 〈 0. 05） at 12 and 24 h after ROSC. Conclusions The lung immune dysfunction induced by imbalance between transcription factors GATA-3 mRNA and T-bet mRNA expressions may complicate in the process of post-resuscitation lung injury in a porcine model of cardiac arrest.

关 键 词：心肺复苏 急性肺损伤 THL Th2失衡 免疫功能障碍 GATA-3 T—bet 

分 类 号：R563.8[医药卫生—呼吸系统]