微囊化异种坐骨神经移植对大鼠脊髓损伤后生长相关蛋白43和神经丝蛋白200表达的影响  

Effects of xenotransplantation of microencapsulated sciatic nerve on the expression of growth-associated protein 43 and neurofilament protein 200 after spinal cord injury in rats

作　　者：刘德明[1] 马学强[2] 

机构地区：[1]南昌大学医学院解剖学教研室,江西省南昌市330006 [2]浙江省台州市立医院外科,浙江省台州市318000

出　　处：《中国组织工程研究与临床康复》2007年第48期9700-9704,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30060034)~~

摘　　要：目的:利用海藻酸钠微囊可以抑制神经移植后免疫排斥反应的特点,将家兔坐骨神经组织细胞微囊化后移植到大鼠脊髓损伤处,观察其对脊髓神经再生的影响。方法:实验于2003-01/2004-06在南昌大学医学院完成。①坐骨神经组织细胞的微囊化:实验无菌条件下取家兔双侧坐骨神经制成组织细胞悬液,与1.5%海藻酸钠溶液混合并喷入20mmol/L的氯化钡溶液中制成微囊化坐骨神经组织细胞;同法制备不含神经组织细胞的海藻酸钡空囊。②实验动物:80只成年Sprague-Dawley(SD)大鼠随机分为正常组、对照组、空囊组和微囊组,其中正常组5只,其他每组25只。③实验方法:SD大鼠对照组、空囊组和微囊组大鼠在脊髓半横断伤后,立即于损伤处分别植入明胶海绵、明胶海绵吸附的10μL空囊以及明胶海绵吸附的10μL微囊化坐骨神经组织细胞。④实验评估:分别于术后1,3,7,14,28d(每个时相5只大鼠)取出损伤部位脊髓标本;正常组大鼠则取相应节段脊髓。石蜡包埋后切片,行免疫组织化学染色观察生长相关蛋白43和神经丝蛋白200的表达情况。结果:80只大鼠均进入结果分析。脊髓损伤后第1天生长相关蛋白43免疫组化染色即出现阳性,呈棕黄色细小斑点状,以后逐渐增多、增大,第14天时表达至高峰,微囊组与对照组、空囊组比较差异显著(P<0.01);至第28天时则减少,但微囊组仍高于对照组、空囊组(P<0.05)。随时间的推移,神经丝蛋白200阳性神经元的体积密度、数密度和表面积密度均增加。表面积密度在第7天时、体积密度和数密度在第14天时,微囊组与对照组、空囊组比较差异有显著性(P<0.05);到第28天时微囊组的体积密度、数密度和表面积密度都高于另两组(P<0.01)。结论:微囊化异种坐骨神经组织细胞移植对脊髓损后生长相关蛋白43和神经丝蛋白200表达有促进作用。AIM： Sodium alginate microcapsule could inhibit the immunological rejection after nerve transplantation. In this study, the effects of transplantation of microencapsulated sciatic nerve tissue/cells from rabbits on the regeneration of hemisected spinal cord in adult rats were observed.METHODS： The experiment was carried out in the Medical College of Nanchang University from January 2003 to June 2004. （1） Bilateral sciatic nerves of rabbits were dissected under sterile conditions and made into nerve tissue/cell suspension which were mixed with 1.5% sodium alginate solution. The mixture was then extruded using a droplet generation device into 20 mmol/L barium chloride solution to form barium-alginate capsules. Empty microcapsules without nerve tissue/call were made using the same method. （2）A total of 80 adult Sprague-Dawiey rats were randomly divided into normal group, control group, empty microcapsule group and microcapsule group with 5 rats in normal group and 25 in the other groups. （3）The rats in control group, empty microcapsule group and microcapsule group were performed a lateral hemisection injury of spinal cord at left T10 level. The injured sites then were implanted with gelatin sponge, gelatin sponge with 10 μL empty microcapsule and 10 μL microencapsulated sciatic nerve tissue and cells. The thoracic cord of injured site and spinal cord of the normal rats at corresponding level were removed at 1, 3, 7, 14, 28 days after operation （5 rats at each time phase）, postfixed and embedded in paraffin. （4）The sections were stained immunohistochemically for growth-associated protein 43 and neurofilament protein 200. RESULTS： Eighty rats were involved in the result analysis. At the first day after spinal cord injury, the growth-associated protein 43 positive reaction was observed, presenting buffy tiny mottling, and rose to its peak on the 14^th day, which in microcapsule group was remarkably superior to that in control group and empty microcapsule group （P 〈 0.01）. The expressio

关 键 词：微囊 脊髓损伤 异种移植 生长相关蛋白43 神经丝蛋白 

分 类 号：R318.08[医药卫生—生物医学工程]