摘要
目的观察调节性树突细胞（DCregs）对烫伤所致全身炎症反应及动物早期死亡率的影响。方法采用MiniMACS免疫磁性分离系统从100只正常BALB／c小鼠脾脏分离纯化DCregs，以获得其亚群CDllc^low CD45RB^hugh DCs。制备15％总体表面积Ⅲ度烫伤小鼠模型。取20只小鼠，烫伤即刻腹腔注射不同剂量DCregs（1×10^5／ml、5×10^5／ml、10×10^5／ml），观察动物48h死亡率，以确定最佳DCregs干预剂量。另取70只小鼠，按随机数字表法分为正常对照组（7只）、假烫伤组（21只）、烫伤组（21只）和DCregs治疗组（21只）。烫伤组伤后即刻腹腔注射林格液1ml抗休克治疗；DCregs治疗组则给予含10×10^5／ml CDllc^low CD45RB^hugh DCs的林格液1ml治疗。分别于烫伤后12、24、48h无菌取血，采用流式细胞术检测树突细胞的表型表达及血浆炎症因子水平。结果10×10^5／mlDCregs治疗组小鼠死亡率较烫伤组及1×10^5／ml、5×10^5／mlDCregs治疗组均显著下降（0％比80％、80％、60％，均P〈0．01）。10×10^5／mlDCregs治疗组小鼠伤后12、24、48h血浆自细胞介素-6（IL-6）水平（ng／L：98．76±10．02、57．83±6．83、13．29±1．07）显著低于烫伤组（156．32±12．85、84．50±9．29、23．04±2．53，均P〈0．01），血浆巨噬细胞趋化蛋白-1（MCP-1）水平（ng／L：102．79±9．88、42．56±5．90、12．96±1．34）显著低于烫伤组（168．23±23．85、83．39±8．41、42.92±4．96，均P〈0．01）．血浆肿瘤坏死因子-α（TNFα）水平（ng／L：16．84±1．92、16．62±1．28、10．26±1．10）亦显著低于烫伤组（24．16±4．93、24．25±4．01、17．91±1．82，均P％0．01）。结论严重烫伤后腹腔注射脾脏DCregs可能通过有效控制促炎因子的产生而明显降低动物早期死亡率。
Objective To observe the effect of regulatory dendritic cells （DCregs） on burn injury induced proinflammatory eytokine production and mortality rate after a single intraperitoneal injection of CDllc^low CD45RB^hugh DCs to injured mice. Methods DCregs were isolated and purified from spleen of 100 normal BALB/c mice to procure CDllc^low CD45RB^hugh DCs by MiniMACS. Mice were subjected to a 15% total body surface area （TBSA） burn injury on the back. Twenty mice were used, and splenic DCregs （1×10^5/ml, 5 ×10^5/ml, 10 ×10^5/ml） were given to them to investigate the protective effect of DCregs against lethality at postburn hours （PBH） 48, and to decide the optimal dosage of intervention. Another group of 70 mice were used, and they were divided into normal control group （n= 7）, sham burn injury group （n=21）, burn injury group （n=21）, and DCregs treatment group （n=21）. The mice in burn injury group received intraperitoncally 1 ml of Ringer solution for resuscitation. 10×10^5/ml of CDllc^low CD45RB^hugh DCs were added to lactated Ringer solution for intraperitoneal injection in DCregs treatment group. Seven animals of each group were sacrificed at PBFI 12, 24 and 48, respectively, and blood samples were collected aseptically for measurement of cytokine levels in plasma and phenotype expressions on DCs by flow cytometry. Results Treatment with 10×10^5/ml DCregs showed a significant decrease in mortality rate compared with burn injured mice and burn injured mice given lower doses of DCregs （1 ×10^5/ml, 5 ×10^5/ml DCregs, 0% vs. 80%, 800% and 600%, all P〈0.01）. A single intraperitoneal injection of 10 ×10^5/ml DCregs to burn injury mice showed a significant decrease in plasma interleukin-6 （IL-6, ng/L; 98.76 ± 10.02, 57.83±6.83, 13.29± 1.07） compared with burn injury mice （156.32± 12.85, 84.50± 9.29, 23.04±2. 53） at PBH 12, 24 and 48 （all P〈0.01）. Similarly, in 10×10^5/ml DCregs treatment group, plasma macrophage chemoattractant protein-1 （MCP-1） levels （ng/L; 102.79 ± 9.88, 42.56 ± 5.90, 12.96±1.34） were markedly lower than those in burn injury group （168.23±23.85, 83.39±8.41, 42. 92± 4.96） at PBH 12, 24 and 48 （all P〈0.01）. A single intraperitoneal injection of 10×10^5/ml DCregs to burn injury mice showed significant reduction in plasma tumor necrosis factor-α （TNF-α） levels （ng/L; 16.84± 1.92, 16.62±1.28, 10.26±1.10） compared with burn injury mice （24.16±4.93, 24.25±4.01, 17.91± 1.82） at PBH 12, 24 and 48 （all P〈0. 01）. Conclusion DCregs may effectively improve the outcome of mice with severe burn injury through a single excessive inflammatory reaction.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2011年第5期279-282,共4页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目（81000847,81071545）
关键词
树突细胞
调节性
烫伤
脓毒症
炎症反应
细胞因子
Regulatory dendritic cell
intraperitoneal injection of DCregs accompanied by lowering Burn injury
Sepsis
Inflammatory response
Cytokine
