地塞米松对人胚神经干细胞移植治疗大鼠脑液压损伤的影响  

Influence of dexamethasone on human embryonic neural stem cell transplantation for rat fluid percussion brain injury

作　　者：龙新兵[1] 张泽舜[1] 韩富[1] 万虹[2] 

机构地区：[1]广州中医药大学第二附属医院神经外科,广东省广州市510120 [2]北京市神经外科研究所,北京市100050

出　　处：《中国组织工程研究与临床康复》2007年第33期6633-6636,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:有较多的学者提出,中枢神经系统是一个部分免疫豁免区,仍然存在干细胞移植后的排异问题,为求证免疫排异的存在与否,实验拟探讨人胚神经干细胞移植治疗脑液压损伤大鼠的移植免疫及其对策。方法:实验于2003-01/2003-07在北京神经外科研究所神经干细胞室完成。①实验材料:人胚来源:12周龄死胎大脑,产妇及家属知情同意,来自北京天坛医院产科。SD雌性大鼠30只,体质量(290±10)g,购自中国医学科学院实验动物研究所。②实验方法:体外培养人胚神经干细胞,并用5-溴脱氧尿嘧啶(BrdU)标记,液压冲击大鼠右侧大脑皮质运动感觉区制作脑损伤模型,将其分为实验组与对照组。实验组在移植人胚神经干细胞后使用地塞米松腹腔注射,对照组移植人胚神经干细胞后仅使用生理盐水腹腔注射。③实验评估:脑外伤后24h移植人胚神经干细胞,于人胚神经干细胞移植后1周脑组织切片免疫组织化学染色检测BrdU、巢蛋白、微管相关蛋白2和胶质纤维酸性蛋白蛋白表达。采用流式细胞仪计数分析检测大鼠脑组织CD4+T细胞淋巴细胞占总淋巴细胞的百分比。结果:两组移植后1周,均见BrdU阳性细胞从移植区域向周围扩散,切片巢蛋白、微管相关蛋白2和胶质纤维酸性蛋白均为阳性,且微管相关蛋白2阳性细胞较多;实验组侧脑室脉络丛和脑实质的微血管中只见有少量的BrdU阳性细胞,对照组脉络丛中见大量聚集的BrdU阳性细胞。两组CD4+T淋巴细胞占总淋巴细胞的百分比差异有显著意义(P<0.05)。结论:异种神经干细胞移植治疗脑损伤可能存在免疫排斥反应,地塞米松对其存在抑制作用。AIM： Many scholars believed that central nervous system （CNS） was a partial immune-privileged sites and rejection was found after stem cell transplantation. This article is designed to investigate the transplantation immunity and strategy of human embryonic neural stem cell transplantation for fluid percussion brain injury in rats, so as to verify the existence of immune rejection. MF, THODS： The experiment was performed at Laboratory of Neural Stem Cells of Beijing Neurosurgery Institute form January to July 2003. ①Human embryo was obtained form 12-weeks-old dead foetus at Department of Obstetrics of Beijing Tiantan Hospital that were obtained permission of puerperants and their family. Totally 30 SD female rats with the body mass of （290±10） g were purchased in the Animal Center of Chinese Academy of Medical Science. ②Human embryonic neural stem cells （HNSCs） were cultured in vitro, labeled with 5-bromodeoxyuridine （BrdU）, and transplanted into rat right brain area injured by fluid percussion. The injured rats were divided into experimental group and control group. Dexamethasone was applied by intraperitoneal injection in the experimental group after HNSCs transplantation, but only saline was applied in the control group. ③HNSCs were implanted 24 hours after cerebral trauma. Immunohistochemical method was used to detect the expressions of BrdU, nestin, microtubule-associated protein （MAP） 2 and glial fibrillary acidic protein （GFAP） 1 week after HNSCs transplantation. The percentage of CD4^＋T lympholeukocyte to total lympholeukocyte was determined by flow cytometry. RESULTS： One week after HNSCs transplantation, BrdU positive cells could be seen and the expression of nestin, MAP2 and GFAP was detectable simultaneously in the same position. Moreover, MAP2 expressed more. A few BrdU positive cells appeared in lateral plexus and capillary of brain substance in the experimental group, and a mass of BrdU positive cells appeared in lateral plexus of the control group. There was

关 键 词：神经干细胞 颅脑损伤 移植 地塞米松 移植免疫 

分 类 号：R651.15[医药卫生—外科学]