血液透析滤过对多器官衰竭犬内质网应激介导的肝细胞凋亡的影响  

The effect of hemodiafiltration on liver cell apoptosis mediated by the endoplasmic reticulum stress signaling pathways in dogs with multiple organ dysfunction syndrome

作　　者：顾凤娟 隋晓露 许云鹏[1] 张艾莎 陈继红[1] 

机构地区：[1]新疆医科大学第一附属医院肾内科,乌鲁木齐830054

出　　处：《新疆医学》2017年第8期835-838,共4页Xinjiang Medical Journal

基　　金：国家自然科学基金(81360121)

摘　　要：目的观察连续血液透析滤过(CVVHDF)治疗多器官功能障碍综合征(MODS)对肝脏功能的影响,并探讨其对肝脏细胞内质网应激介导的细胞凋亡影响。方法 14只Beagle犬采用失血性休克+复苏灌注+内毒素血症^([1])建立犬MODS模型^([2-3]),分为MODS组(n=6)和CVVHDF组(n=8)。内毒素注射完毕后CVVHDF组接受CVVHDF治疗24 h,MODS组不接受CVVHDF治疗。术前(T1)、内毒素注射完后0h(T2)、6h(T3)、12h(T4)及24h(T5)留血标本。荧光定量PCR法检测肝细胞CHOP、JNK、Caspase-12mRNA基因表达水平。Western blot法测定肝细胞CHOP、JNK、Caspase-12 mRNA蛋白表达。Annexin V-PI染色流式细胞仪检测肝细胞凋亡率。结果与MODS组相比,CVVHDF治疗后,肝脏细胞JNK mRNA在表达增高,差异有统计学意义(P<0.01);CHOP蛋白表达增高,差异有统计学意义(P<0.05);凋亡细胞计数明显降低,差异有统计学意义(P<0.05)。结论 MODS病程中诱发内质网应激,激活下游凋亡信号通路,CVVHDF治疗后减弱内质网应激激,减轻肝脏细胞凋亡。Objective To observe the effect of continuous veno-venous hemodiafiltration（CVVHDF）on liver function in multiple organ dysfunction syndrome（MODS） and to discuss the expression of apoptosis mediated by endoplasmic reticulum stress signaling pathways. Methods Two-hit method was used to establish the dog MODS model. Fourteen Beagle dogs were divided into MODS group（n=6）and CVVHDF group（n=8）. CVVHDF was performed about 24 hours after LPS injection in MODS dogs of CVVHDF group. Blood samples before operation（T1）,before LPS injection and 0h（T2）,6h（T3）,12h（T4）,24h（T5）after LPS injected were collected. L liver cell CHOP,JNK,Caspase-12 gene expression levels were detected by Fluorescence quantitative PCR method;Western blot method was used to determine liver cell CHOP,JNK,Caspase-12 mRNA expression. Annexing V-PI staining flow cytometry was used to detect liver cell apoptosis. Results The levels of JNK mR NA levels and CHOP protein expression in liver were higher in CVVHDF group than in MODS group（P0.01）;The count of apoptotic cells had a significant difference between two groups（P0.05）. Conclusion There were three signaling pathways mediated apoptosis in MODS dogs over endoplasmic reticulum stress,CVVHDF can reduce the endoplasmic reticulum stress and reduce apoptosis.

关 键 词：连续血液透析滤过 多器官功能障碍 内质网应激 肝脏 

分 类 号：R457[医药卫生—治疗学]