Bruton酪氨酸激酶和核因子-κB在高氧诱导肺损伤中的作用  

Effect of Bruton＇ s tyrosine kinase in hyperoxia-induced acute lung injury

作　　者：孟辉[1] 曹大伟[1] 刘超锋[1] 侯明霞[1] 何方[1] 张新日[1] 

机构地区：[1]山西医科大学第一医院呼吸科,太原030001

出　　处：《中华急诊医学杂志》2017年第2期181-185,共5页Chinese Journal of Emergency Medicine

基　　金：山西省科技成果推广计划项目（20140710）;山西省自然科学基金（2012011037-2）

摘　　要：目的观察Bruton酪氨酸激酶（ Bruton＇ s tyrosine kinase, Btk）和核因子-κB（ nuclearfactor kappa B, NF-κB） 在高氧诱导的急性肺损伤（hyperoxia-induced acute lung injury, HALI）小鼠肺组织中的表达水平，探讨两者在HALI发生中的作用机制。方法雄性昆明小鼠72只，按随机数字表法随机分为四组：对照组、高氧暴露3d组（H3d组）、高氧暴露3d＋抑制剂组（H3d＋I组）和抑制剂组。光镜下观察各组小鼠肺组织病理学改变，测定支气管肺泡灌洗液（BALF）总蛋白含量（TP）和肺湿／干质量比值（W／D）；采用Westernblot法测定肺组织Btk、p-Btk和pNF-κBp65蛋白表达；采用RT—qPCR法测定肺组织白细胞介素-6（IL-6）mRNA表达，采用ELISA法检测血清中单核细胞趋化因子-1（MCP-）含量。各组间比较采用单因素方差分析。结果对照组与抑制剂组比较，各测定数据差异均无统计学意义（P〉0．05）。与H3d＋I组比较，H3d组肺组织病理损伤明显增加且BALF总蛋白含量和W／D比值均明显增高，（分别为P=0．002，P=0．000）差异均具有统计学意义；H3d组肺组织Btk、p-Btk和pNF—κBp65表达量明显高于H3d＋I组（分别为P=0．002。P=0．013。P=0．000）。H3d组肺组织IL-6mRNA表达水平明显高于对照组（P=0．004）、抑制剂组（P=0．000）和H3d＋I组（P=0．021）；H3d组血清MCP-1含量均高于对照组（P=0．002）、抑制剂组（P=0．000）和H3d＋I组（P=0．009）。相关性分析结果显示，各组小鼠肺组织Btk或p-Btk表达水平与pNF—κBp65表达量之间呈正相关（r=0．902；r=0．954，P〈0．01）。结论Btk可能通过调控NF-κB信号通路而介导IL-6和MCP-1等炎症细胞因子释放，在HALI发牛中起首要作用．抑制Btk的活性可有效减轻肺损伤的严重程度。Objective To investigate both in mechanism of hyperoxia-induced acute lung injury （HALI） by vivo experiment, to observe the Bruton＇ s tyrosine kinase （Btk） and nuclear factor kappa B （NF-κB） signals expression level. Methods Total of 72 healthy male Kunming mice were randomly （random number） divided into four groups： air control group, hyperoxia exposure 3 days group （H3d group） , hyperoxia exposure 3 days ＋ inhibitor group （H3d ＋ I group） and inhibitor groups . Then the pathological changes of lung tissues were observed under light microscope; The total protein content （TP） of bronchoalveolar lavage fluid （BALF） and wet/dry weight ratio （W/D） of lung were detected; The protein expression of Btk, p-Btk, pNF-κB p65 were mersured by Western blot; the mRNA level of IL-6 was determined by real-time polymerase chain reaction （qRT-PCR）; the level of monocyte chemoattractant protein-1 （MCP-1） in serum was detected by enzyme-linked immunosorbent assay （ELISA） . Statistcal significance was determined by 1-way ANOVA. Results There were no significant difference in the data between the control group and the inhibitor group （ P 〉 0. 05 ） . The pathological injury in light microscope, content of total protein in BALF, W/D ratio of lung tissues in H3d group were significantly higher than H3d ＋ I group （ Respectively P = 0. 002, P = 0. 000 ） . Western blot analysis showed that expression of Btk, p-Btk, pNF-κB p65 in H3d group were significantly higher than those in H3d ＋ I group （ Respectively P =0. 002, P=0. 013, P =0. 000） . RT-qPCR results showed that the expression of IL-6 mRNA in H3d group were significantly higher than control group （P =0. 004）, inhibitor group （P =0. 000） and H3d ＋ Igroup （P =0. 021 ） . In addition, The serum MCP-1 levels in H3d group were higher markely than the control group （P =0. 002） , inhibitor group （P =0. 000） and H3d ＋ I group （P =0. 009） . The correlation analysis showed that pNF-κB p65 wer

关 键 词：高氧 急性肺损伤 Bruton酪氨酸激酶 核因子-ΚB 小鼠 湿/干质量比值 单核细胞趋化因子-1 信号通路 

分 类 号：R563.8[医药卫生—呼吸系统]