骨髓间充质干细胞移植对创伤性脑损伤大鼠肿瘤坏死因子α、核转录因子κB因子表达的影响  

Effect of bone marrow mesenchymal stem cells transplantation on expression of TNF-a and NF-kB in rats following traumatic brain injury

作　　者：王斌[1,2] 崔建忠[3] 王勇朝[1,2] 张娟[1,2] 李冉[1,2] 田艳霞[1,2] 赵曼曼[1,2] 高俊玲[1,2] 

机构地区：[1]河北联合大学基础医学院组织学与胚胎学教研室 [2]河北省煤矿卫生与安全实验室,唐山063000 [3]唐山市工人医院神经外科,唐山063000

出　　处：《解剖学杂志》2013年第5期929-932,951,共5页Chinese Journal of Anatomy

基　　金：河北省自然科学基金（H2012401071）

摘　　要：目的：观察肿瘤坏死因子a（TNF-a）、核转录因子”B（NF-kB）在大鼠弥漫性脑创伤后脑组织的表达变化，并进一步探讨骨髓间充质干细胞（BMSCs）对其变化的影响。方法：分离、培养SPF级雄性5周龄SD大鼠BMSCs用于实验；SPF级成年雌性SD大鼠60只随机分为假手术组（Sham组），创伤性脑损伤（TBI）组，BMSCs治疗组。采用Marmarou法制成大鼠创伤性脑损伤模型。神经功能评分（mNSS）观察大鼠神经行为表现；免疫组织化学检测TNF—a、NF-kB表达及定位；免疫印迹检测皮质TNF-a、NF-kB的蛋白定量；免疫荧光检测性别决定基因SRY在脑组织中的分布情况。结果：BMSCs治疗组大鼠脑创伤后的神经功能明显改善，与TBI组相比差异具有统计学意义。与Sham组相比，TBI组TNF-u、NF-xB蛋白表达明显上调，TNF-a、NF-kB在伤后1d增加，3d达到高峰，7～14d渐回落，仍高于基线水平；BMSCs治疗组TNF-a、NF-kB的表达均低于TBI组。激光共聚焦显微镜观察到损伤区脑皮质中SRY的表达。结论：脑外伤早期TNF-a、NF-kB即被激活，BMSCs对脑创伤大鼠具有脑保护作用，其机制之一是通过调节NF-kB、TNF-a的表达而实现的。Objective： To investigate the expression of the tumor necrosis factor-a （TNF-a）, nuclear transcription factor-kB （NF-kB）, and the effect of bone marrow mesenchymal stem cells （BMSCs） on the rats following traumatic brain injury （TBI）. Methods： The cultured BMSCs isolated from SPF male 5-week-old SD rats were used in the experiment; 60 adult female SD rats of SPF level were randomly divided into 3 groups： a sham, TBI, BMSCs treatment groups. The rat model of diffused brain injury was established based on Marmarou＇s method. Immunohistochemical and Western blotting were used to determine the expression of TNF-a and NF-kB. Behavioral tests were performed by Modified Neurological Severity Score （mNSS）. The expression of sex-determining gene （sry） were detected by immunofluorescence labeling. Results： Compared with the sham group, TNF-a and NF-kB began to increase in 1 day following TBI, when peaked at 3 day, and then descended from 7 d, but was still higher than the basic level at 14 d. At the corresponding time, compared to the TBI group, the expression of TNF-a and NF-kB decreased remarkably in the BMSCs treatment group. The mNSS were significantly improved in BMSCs treatment group, compared with TBI group. Laser scanning confocal microscopy was used to observe the location of SRY in the cortex of the damage zone. Conclusion： At the early stage of TBI, TNF-a and NF-kB are activated. Bone marrow mesenchymal stem cells have a protective effect against TBI, which may be related to the expression inhibition of NF-kB and TNF-a.

关 键 词：创伤性脑损伤 骨髓间充质干细胞 神经功能评分 肿瘤坏死因子A 核转录因子xB 大鼠 

分 类 号：R542.22[医药卫生—心血管疾病]