人脐带间充质干细胞对脓毒症大鼠的免疫干预研究  

Immune intervention of human umbilical cord mesenchymal stem cells on sepsis rats

作　　者：张何为[1] 崔晓旭[3] 方涛[3] 傅强[2] 

机构地区：[1]天津医科大学,300070 [2]天津市第四中心医院中心实验室,300140 [3]天津市第四中心医院重症医学科,300140

出　　处：《中华危重病急救医学》2017年第8期705-710,共6页Chinese Critical Care Medicine

基　　金：天津市“131”创新人才培养项目（2012）

摘　　要：目的 探讨人脐带间充质干细胞（UC-MSCs）对脓毒症大鼠免疫细胞及炎性因子的调节作用.方法 按照随机数字表法将184只雄性SD大鼠分为正常对照组（n=8）、假手术组（n=48）、脓毒症模型组（n=64）、干细胞治疗组（n=64）.采用盲肠结扎穿孔术（CLP）制备脓毒症动物模型.干细胞治疗组于制模后1 h腹膜注射第3代UC-MSCs（2×106/mL）1 mL;假手术组、脓毒症模型组给予等量生理盐水.假手术组、脓毒症模型组、干细胞治疗组各取16只大鼠观察72 h内存活情况;其余大鼠分别于术后12、24、48和72 h各取8只收集门静脉血,用流式细胞仪检测CD4＋T细胞百分比及其辅助性T细胞（Th1/Th2）比值,用酶联免疫吸附试验（ELISA）测定血浆肿瘤坏死因子-α（TNF-α）、高迁移率族蛋白B1（HMGB1）、白细胞介素-10（IL-10）水平.结果 干细胞治疗组术后72 h大鼠存活率略高于脓毒症模型组〔62.5%（10/16）比50.0%（8/16）,χ2=0.509,P〉0.05〕.脓毒症模型组术后12 h CD4＋T细胞百分比、Th1/Th2比值均显著高于假手术组,随时间延长有所降低,并于48 h后显著低于假手术组;血浆炎性因子水平均于术后12 h显著高于假手术组,TNF-α、IL-10于术后48 h达高峰后有所降低,而HMGB1持续升高至72 h.与脓毒症模型组比较,干细胞治疗组术后12 h、24 h CD4＋T细胞百分比显著降低〔（49.66±0.91）%比（59.11±1.17）%,（41.80±0.89）%比（49.84±0.99）%〕,术后12、24、48 h Th1/Th2比值显著降低（0.745±0.065比1.254±0.115,0.407±0.077比0.806±0.061,0.280±0.057比0.454±0.049）及术后12、24、48、72 h TNF-α和HMGB1显著降低 〔TNF-α（ng/L）：52.60±6.60比58.03±6.53,71.77±8.48比147.39±11.37,111.83±10.76比271.36±19.04,83.09±7.43比171.04±14.06;HMGB1（ng/L）：149.12±9.89比187.33±12.79,192.94±14.92比442.35±52.72,1393.67±88.86比1950.90±126.66,1875.84±111.67比2557.12±186.01〕,术后12、24、48、72 h IL-10水平显著Objective To investigate the effect of human umbilical cord mesenchymal stem cells （UC-MSCs） on immune cells and inflammatory factors in septic rats.Methods 184 male Sprague-Dawley （SD） rats were divided into normal control group （n = 8）, sham operation group （n = 48）, sepsis model group （n = 64）, and UC-MSCs treatment group （n = 64）. An animal model of sepsis was produced by cecal ligation and puncture （CLP）. In the UC-MSCs treatment group 1 mL UC-MSCs （2×106/mL） were injected intraperitoneally at 1 hour after the model establishment;the sham operation group and the sepsis model group were given the same amount of saline. Sixteen animals in each group of the sham operation group, sepsis model group, and UC-MSCs treatment group were observed for 72-hour survival rate. The percentages of CD4＋ T cells and the ratio of helper T cells 1/2 （Th1/Th2） in whole blood cells were measured by flow cytometry at 12, 24, 48 and 72 hours after operation. The levels of tumor necrosis factor-α （TNF-α）, high mobility group box 1 （HMGB1）, interleukin-10 （IL-10） were measured by enzyme linked immunoabsorbent assay （ELISA）.Results The 72-hour survival rate of the UC-MSCs treatment group was slightly higher than that of the sepsis model group [62.5% （10/16） vs. 50.0% （8/16）,χ2 = 0.509,P 〉 0.05]. The percentage of CD4＋ T cells and Th1/Th2 ratio in the sepsis model group were significantly higher than those in the sham operation group at 12 hours after operation, and decreased as the time prolonged to 48 hours. The levels of plasma inflammatory factors were significantly higher than those of sham operation group at 12 hours after operation, TNF-α and IL-10 were decreased at 48 hours after operation, while HMGB1 continued to increase until 72 hours after operation. Compared with those in the sepsis model group, the percentages of CD4＋ T cells at 12 hours and 24 hours after operation [（49.66±0.91）% vs. （59.11±1.17）%, （41.80±0.89）% vs. （49.84±0.99）

关 键 词：脓毒症 脐带间充质干细胞 T淋巴细胞 炎性介质 

分 类 号：R459.7[医药卫生—急诊医学]