HBO联合BMSCs移植减轻脊髓损伤大鼠炎症反应与NF-KB炎症信号通路有关  

Hyperbaric oxygen combined with bone marrow mesenchymal stem cells transplantation reduced inflammatory reaction in rats with spinal cord injury and its relation with nuclear factor-kappa B signal pathway inhibition

作　　者：黄玉琴[1] 李萌[1] 张晓菁[1] 张慧[1] 陈银海[1] 

机构地区：[1]南方医科大学珠江医院神经康复及高压氧科,广州510282

出　　处：《中华神经医学杂志》2017年第8期777-784,共8页Chinese Journal of Neuromedicine

基　　金：广东省科技计划项目（20108080701020）

摘　　要：目的探讨高压氧（HBO）联合骨髓间充质干细胞（BMSCs）移植减轻脊髓损伤（SCI）大鼠炎症反应的机制是否与核因子-κB（NF-κB）炎症信号通路有关。方法成年雄性SD大鼠160只按随机数字表法分为假手术组、SCI组、BMSCs组、BMSCs＋HBO组，每组40只。后3组应用改良Allen打击法损伤脊髓制备SCI模型。且BMSCs组、BMSCs＋HBO组分别于建模1h后尾静脉注射1mL BMSCs（3×10^6个）悬液或BMSCs悬液注射＋HBO治疗。伤后1、3、7、14d时采用BBB评分评估各组大鼠后肢运动功能；伤后3d时采用Westem blotting检测各组大鼠脊髓组织NF-KB及其抑制蛋白（IKB-α）、肿瘤坏死因子-α（TNF-α）、白介素-1β（IL^-1β）蛋白含量，采用ELISA法检测脊髓组织TNF-α、IL^-1β浓度，采用免疫组化染色检测脊髓组织NF-KBp65阳性细胞，采用HE染色观察脊髓组织病理学改变。结果BMSCs组及BMSCs＋HBO组BBB评分在伤后3d、7d、14d时均较SCI组明显升高，其中以BMSCs＋HBO组升高更显著，差异均有统计学意义（P〈0．05）。伤后3d时，BMSCs组脊髓组织NF-KB、TNF-α及IL^-1β蛋白含量较SCI组明显降低，IKB-α蛋白含量明显增高，差异均有统计学意义（P〈O．05）；BMSCs＋HBO组脊髓组织NF-KB、TNF-α及IL^-1β蛋白含量较BMSCs组明显降低，IKB-α蛋白含量明显增高，差异均有统计学意义（P〈0．05）。BMSCs组脊髓组织TNF-α、IL^-1β浓度较SCI组明显减少，差异均有统计学意义（P〈0．05）；BMSCs＋HBO组脊髓组织n师吨、IL^-1β浓度与BMSCs组比较明显减少，差异均有统计学意义（p〈0．05）。BMSCs组NF-kB p65阳性细胞数较SCI组明显降低，差异有统计学意义（P〈0．05）；BMSCs＋HBO组NF-KBp65阳性细胞数与BMSCs组比较明显减少，差异有统计学意义（P〈0.05）。与SCI组比较，BMSCs组、BMSCs＋HBO组炎症反应、水肿减轻，脊髓空洞面积减少，损伤区域结构改善。结论HBO联合BMSObjective To explore the mechanism of hyperbaric oxygen （HBO） combined with bone marrow mesenchymal stem cells （BMSCs） transplantation to reduce the inflammatory response of spinal cord injury （SCI） in rats, and whether it is related to nuclear factor-kappa B （NF-KB） signaling pathway or not. Methods One hundred and sixty male adult SD rats were randomly divided into 4 groups according to the random number table： sham-operated group, SCI group, BMSCs transplantation group and BMSCs Iransplantation＋HBO group （n=40）. The rats from the latter three groups were established the SCI models by modified Allen dropping; and 1 mL BMSCs （3×10^6 cells） suspension was given by tail vein injection one h after SCI into the rats of the BMSCs transplantation group and BMSCs transplantation＋HBO group; rats in the BMSCs transplantation＋HBO group were given hyperbaric oxygen treatment. Rats were evaluated for locomotor function of hind limbs with Basso-Beattle-Bresnahan （BBB） scale one, 3, 7, 14 d after SCI. Three d after SCI, the expression levels of NF-KB, IKB-α, tumor necrosis factor （TNF）-α and interleukin （IL）-1β in the spinal cord tissues were determined by Western blotting; and enzyme-linked immuno sorbent assay （ELISA） was employed to detect the TNF-α and IL^-1β levels. The number of NF-KB p65 positive cells in the spinal cord tissues was determined by immunohistochemistry. The histopathological changes of injured spinal cords were observed by HE staining. Results BBB scores of BMSCs transplantation group and BMSCs transplantation＋HBO group were significantly higher than those of SCI group 3, 7 and 14 d after SCI （P〈0.05）, and those in the BMSCs transplantation＋HBO group increased more significantly. The contents of NF-KB, TNF-α and IL^-1 β in the spinal cord tissues of BMSCs transplantation group were significantly lower than those in the SCI group 3 d after SCI, and the content of IKB-α protein was significantly increased （P〈0.05）; the contents of NF-

关 键 词：脊髓损伤 高压氧 骨髓间充质干细胞 核因子-KB 炎症反应 

分 类 号：R651.2[医药卫生—外科学]