沙漠干热环境猪创伤失血性休克肾损伤氧化应激及Caspase-3变化研究  

The changes of oxidative stress and caspase-3 in swine with traumatic hemorrhagic shock in dry-heat environment of desert

作　　者：安宗仁 李新宇[2] 刘江伟[2] 沈才福 段越[2] 李建瑛[2] 李佳佳[2] 

机构地区：[1]石河子大学医学院,石河子832000 [2]新疆军区总医院新疆特殊环境医学重点实验室,乌鲁木齐830000

出　　处：《中华急诊医学杂志》2017年第5期522-527,共6页Chinese Journal of Emergency Medicine

基　　金：总后勤部面上基金项目（CLZ13J003）

摘　　要：目的研究PI3K/Akt/GSK3β信号通路在颈部降温的调节作用，初步探讨颈部降温的脑保护作用是否与PI3K/Akt及GSK3β的磷酸化有关。方法实验在第二军医大学附属长征医院急救科实验室完成。健康雄性新西兰家兔30只随机（随机数字法）分为5组，采用4 min室颤模型。（1）假手术组：常规手术操作置管，不诱导CA，于术后24 h处死取标本。（2）常温复苏组（normothermia theat，NT组）：常规致颤复苏并于24 h时间点处死动物取标本。（3）复苏即刻降温组（intra-arrest therapeutic hypothermia，IATH组）：于心肺复苏同时启动颈部快速降温，目标脑温为34 ℃，以后维持目标脑温至ROSC后4 h。于24 h时间点处死动物取标本。（4）复苏中降温＋特异性抑制剂LY294002（LY294002组）：复苏前20 min脑室注射LY294002，其余同组3。（5）复苏后1 h降温组（post-arrest therapeutic hypothermia，PATH组），于心肺复苏后1 h启动颈部快速降温，目标脑温为34 ℃，其余同组3。LY294002溶于DMSO稀释成10 μmol/L，在动物脑立体定位仪下在ROSC前20 min前给予脑室内注射，其余各组给予溶剂DMSO。动物24 h过量麻醉处死，采集标本。应用Western blot检测Akt、p-Akt、GSK-3β、P-GSK-3β （ser9）的蛋白表达，运用TUNEL等方法观察各组组织凋亡变化。所得数据以均数±标准差（±s）表示，多组间比较采用单因素方差分析。结果与Sham组比较，常温心肺复苏组兔脑细胞胞质中的Akt （Thr-308）磷酸化水平（P-AKT）和P-GSK3β在心肺复苏后24 h明显减低（P〈0.05）;复苏即刻颈部降温组P-AKT和P-GSK3β水平在心肺复苏后24 h均较常温复苏组明显增强（P〈0.05）；复苏后1 h降温组的蛋白表达水平在复苏后24 h也较常温复苏组有增强（P〈0.05），但弱于复苏即刻颈部降温组；脑室内注射LY294002去除了亚低温的这一作用，表明了LY294002抑制Akt的磷酸化。复苏�Objective To study the changes of oxidative stress and caspase-3 in swine with traumatic hemorrhagic shock in.dry-heat environment of desert. Methods A total of 48 Landrace small swine were randomly （random number） divided into 2 groups （n = 24 in each group ）, and then the traumatic hemorrhagic shock was established in room temperature environment [ （25 ± 1 ） ℃, （35 ± 5 ） % humidity ] and in dry-heat environment [ （40. 5 ±0. 5）℃ , （ 10 ±2）% humidity] in swine. Dry-heat environment traumatic hemorrhagic shock group （ DHS）, which was made in an artificial experiment cabin mimic the reality included swine exposed in the dry-heat environment of desert for 3 h （ T0, n = 6） , T1 （50 min after shock modeling, n = 6） , T2 （100 min after shock modeling, n = 6） , T3 （150 min after shock modeling, n =6）. At each interval, blood sample was coUected to detect urea nitrogen （BUN） and creatinine, urine sample was collected to detect neutrophil gelatinase-associated lipoprotein （NGAL）, kidney tissue samples were collected to evaluate renal morphological and tubular scores, as well as to detect catalase （ CAT）, superoxide dismutase （SOD） and malondialdehyde （MDA）. Western blot was used to detect the level of easpase-3. Traumatic hemorrhagic shock group of room temperature environment （RTS） was established and variety of assays were carried out as same as those deteced in the dry-heat environment group. Results Compared with the room temperature environment exposed group, kidney damage index, antioxidant and caspase-3 were increased in desert dry-heat environment exposed for 3 h group, but there were no statistically significant difference （P 〉 0. 05 ）. And from T1 then on, the levels of NGAL, CAT and SOD in DHS groups were increased which were significant different from those in RTS group （ P 〈 0. 05 or P 〈 0. 01 ）. There were significant differences in BUN and creatinine at T2 between two groups （ P 〈 0. 05 ）. At T3, caspase-3 prote

关 键 词：沙漠 干热环境 创伤失血性休克 氧化应激 细胞凋亡 急性肾损伤 

分 类 号：R641[医药卫生—外科学]