骨髓间充质干细胞对多器官功能障碍综合征大鼠炎症因子表达的影响  

Effects of bone marrow mesenchymal stem cells on the expressions of inflammatory factors in rats with multiple organ dysfunction syndrome

作　　者：修光辉 周霞 李秀玲[2] 孙洁 陈献忠 陈晓雷 潘兴华 凌斌 Xiu Guanghui;Zhou Xia;Li Xiuling;Sun Jie;Chen Xianzhong;Chen Xiaolei;Pan Xinghua;Ling Bin(ICU Department,the Fourth Affiliated Hospital of Kunming Medical University（the Second People＇s Hospital of Yunnan Province）,Kunming 650021,China;Obstetrics Department,the First People＇s Hospital of Yunnan Province,Kunming 650031,Chin;Clinical Laboratory Department,Kunming General Hospital of Chengdu Military Region,Kunming 650032,China)

机构地区：[1]昆明医科大学第四附属医院(云南省第二人民医院)重症医学科,昆明650021 [2]云南省第一人民医院产科,昆明650031 [3]解放军昆明总医院临床实验科,昆明650032

出　　处：《中华急诊医学杂志》2018年第9期988-994,共7页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金（81360289）;云南省科技厅-昆明医科大学联合专项资金[2017FE468（-180）,2015FB079,2013FB185,2012FB076,2017FE467（-194）];云南省高层次卫生计生技术人才培养经费资助（H-2017060）

摘　　要：目的探讨骨髓间充质干细胞（bonemarrow mesenchymal stem cells，BMSC）对多器官功能障碍综合征（MODS）大鼠炎症因子表达的影响。方法体外培养4周龄SD大鼠BMSC并鉴定成功，取第四代BMSC用于实验，按照随机数字表法将SD大鼠分成3组（每组20只）：即假手术（Sham）组、MODS组和BMSC组。MODS组大鼠经大隐静脉注射脂多糖（LPS，1mg／kg）制作动物模型，Sham组注射等量磷酸盐缓冲液（PBS），BMSC组在造模后2h内经尾静脉输注BMSC（1×10^6／只，0．5mL）。术后72h观察三组大鼠的存活率并绘制生存曲线；采用苏木精．伊红（HE）染色观察肺脏、肝脏、心脏组织的病理学改变；血气分析和生化检测评估器官功能；采用蛋白芯片技术检测34种相关炎症因子，筛选差异表达因子，用酶联免疫吸附试验（ELISA）验证差异表达因子。多组定量资料采用单因素方差分析，两两比较采用LSD-t检验，生存曲线差异的比较采用Log-rank检验，以P〈0．05为差异有统计学意义。结果Sham组、MODS组和BMSC组大鼠生存率分别为100％、60％、80％；生存曲线显示，Sham组的生存率明显大于MODS组及BMSC组（Sham vs．MODS χ^2=9．798，P=0．0017；Sham vs．BMSC χ^2=4．333，P=0．0374）；BMSC组的生存率大于MODS组（BMSC vs．MODS χ^2=2.408，P=0．1207），但差异无统计学意义。HE染色结果显示，MODS组肺脏、肝脏、心脏组织充血水肿，炎症细胞浸润，BMSC组组织水肿与炎症细胞浸润减轻；血气分析及生化指标检测显示，与Sham组比较，MODS组pH、PaCO2、乳酸（Lac）的水平明显升高（均P〈0．01），总胆红素（TB）明显升高f（0．801±0．501）U／Lvs．（2．533±0．382）u／L，P=0．003]及白蛋白（ALB）明显降低[（35．471±4．015）U／L vs．（23．202±4．872）U／L，P〈0．01]，肌酸激酶（CK）明显升高[（315．670±41．402）U／Lvs．（708．250±219．201）U�Objective To investigate the effects of bone marrow mesenchymal stem cells （BMSC） on the expression of inflammatory factors in rats with multiple organ dysfunction syndrome （MODS）. Methods BMSC extracted from the 4-week-old Sprague-Dawley （SD） rats was cultivated and identified in vitro, then the 4th passage of which was used in the experimental study. Sixty SD rats were randomly（random number） divided into three groups （n=20 in each group）： Sham group （SG）, MODS group （MG） and BMSC group （BG）. Rats in the MG was injected by 1 mg/kg lipopolysaccaride （LPS） via great saphenous vein, rats in the SG injected with the same volume sterile phosphate buffer saline and rats in the BG infused by 1×10^6/cells BMSCs through the tail vein at 2 h after LPS injection. The survival rate, tissue pathological changes of the lung, liver and heart by hematoxylin and eosin （HE） staining, organ dysfunction measurement by blood gas analysis and biochemical indicators as well as the related inflammatory factors by protein microarray and enzyme linked immunosorbent assay （ELISA）, were detected 72 h post operation. Multi-group quantitative data was analyzed by one way ANOVA, paired-comparisons by LSD-t test and the comparisons of survival curves in the three groups by Log-rank test. The value of P〈0.05 was considered statistically significant. Results The survival rate in SG, MG and BG was 100%, 60% and 80%, respectively. The survival curves showed that the survival rate of SG was higher than the MG and BG （SG vs. MG, χ^2=9.798, P=0.001 7; SG vs. BG, 3（2=4.333, P=0.037 4）, but there was no significant difference comparing the BG to the MG （χ^2=2.408, P=0.120 7）. The tissue congestion and edema, and inflammatory cells infiltration in the lung, liver, and heart of the MG were observed by HE staining, while these changes reduced in the BG. Compared with the SG, the levels of pH and PaCO2 and lactic acid （Lac） increased significantly （all P〈0.01）, the level of total bilir

关 键 词：多器官功能障碍综合征 骨髓间充质干细胞 干扰素-Γ 单核细胞趋化因子-γ 炎症因子 

分 类 号：R459.7[医药卫生—急诊医学]