高容量血液滤过对多器官功能障碍犬CHOP蛋白的影响  

Effect of high volume hemofiltration on CHOP protein from a canine model of multiple organ dysfunction syndrome

作　　者：努尔孜叶·阿布里克木 廖汉文[2] 王顺[1] 马琴[1] 帕旦木·托乎提 隋晓露 马林[1] 胡磊[1] 陈继红[1] 

机构地区：[1]新疆医科大学第一附属医院肾病科,乌鲁木齐830054 [2]新疆伊犁州友谊医院肾病科,伊宁835100

出　　处：《中华肾脏病杂志》2015年第2期115-119,共5页Chinese Journal of Nephrology

基　　金：基金项目：新疆维吾尔自治区青年科技创新人才培养工程（优秀青年科技创新人才培养项目）

摘　　要：目的观察高容量血液滤过（HVHF）治疗多器官功能障碍综合征（MODS）对CCAAT增强子结合蛋白（CHOP）的影响，并探讨CHOP蛋白在内质网应激介导的凋亡途径中的作用。方法12只Beagle犬采用失血性休克＋复苏灌注＋内毒素血症建立犬MODS模型，随机分为HVHF组（n=6）和MODS组（n=6）。内毒素注射完毕后HVHF组接受HVHF治疗24h，MODS组不接受HVHF治疗。体内实验：留取各时间点（术前、内毒素注射完毕后0h、6h、12h及24h）血清样本，处死时即留取肝、肾、肺组织标本并测定CHOPmRNA表达。体外实验：两组血清体外诱导人脐静脉内皮细胞（HUVECs）建立内质网应激凋亡模型。测定CHOPsiRNA干扰前后CHOPmRNA表达、蛋白表达及内皮细胞凋亡率。结果体内实验结果：两组肝、肾、肺组织CHOPmRNA表达水平的差异无统计学意义（P〉0．05）。体外实验结果：（1）CHOPmRNA表达：与MODS组相比，HVHF组CHOPmRNA表达水平在内毒素注射完毕后6h、24h显著下降，差异有统计学意义（P〈0．05）。与CHOPsiRNA干扰前相比，干扰后两组CHOPmRNA水平各时间点均下降，差异有统计学意义（P〈0．05）。（2）CHOP蛋白表达：与MODS组相比，各时间点HVHF组CHOP蛋白表达水平均显著下降，差异有统计学意义（P〈0．05）。与CHOPsiRNA干扰前相比，干扰后两组CHOP蛋白表达水平均显著下降，差异有统计学意义（P〈0．05）。（3）内皮细胞凋亡率：与术前相比，两组各时间点内皮细胞凋亡率均显著下降，差异有统计学意义（P〈0．05）。与MODS组相比，HVHF组各时间点内皮细胞凋亡率显著下降，差异有统计学意义（p〈0．05）。与siRNA干扰前相比，干扰后两组内皮细胞凋亡率均显著下降，差异有统计学意义（P〈0．05）。结论HVHF治疗MODS过程中内皮细胞凋亡减少可能与抑制CHOPmRNA表达及蛋白合成有关。Objective To observe the effect of high volume hemofiltration （HVHF） on the expression of CCAAT enhancer binding protein（CHOP） during the treatment of multiple organ dysfunction syndrome （MODS）. To investigate the role of CHOP protein act in apoptosis pathway mediated by the endoplasmic reticulum stress. Methods Twelve Beagle dogs were subjected to hemorrhagic shock plus resuscitation and endotixemia to establish MODS model, then they wererandomly divided into two groups： HVHF group （n=6） and MODS group （n=6）. After endotoxin injection completed, the HVHF group received HVHF treatment for 24 hours; MODS group did not receive. Vivo experiments： Blood samples were obtained at different time points（before operation, 0 h, 6 h, 12 h, 24 h after the injection of endotoxin）. The dogs were killed and the tissue samples from lung, liver and kidney were took, then the expression of CHOP mRNA was determined. Vitro experiments： human umbilical vein endothelial cells （HUVECs） were induced by two groups＇ blood samples to establish the apoptosis model. Gene expression, protein quantification and cell apoptosis rate were determined before and after the interference. Results Vivo experiments： The levels of CHOP mRNA from lung, liver and kidney had no significant difference between the two groups （P 〉 0.05）. Vitro experiments： （1）The expression of CHOP mRNA： Compared with MODS group, the expression levels of CHOP mRNA were significantly decreased in HVHF group at 6 h, 24 h after the injection of endotoxin （P 〈 0.05）. Compared with before, the expression levels of CHOP mRNA in the two groups were both significantly decreased after CHOP siRNA interference （P 〈 0.05）. （2）The expression of CHOP protein： Compared with MODS group, the expression levels of CHOP protein were significantly decreased in HVHF group at each time points （P 〈 0.05）. Compared with before, the expression levels of CHOP protein in the two groups were both significantly decreased after CHOP

关 键 词：血液滤过 多器官功能衰竭 细胞凋亡 内质网应激 转录因子 CHOP 

分 类 号：R459.7[医药卫生—急诊医学]