大鼠急性脊髓损伤后内源性垂体腺苷酸环化酶激活肽表达的变化  

Expressions of endogenous pituitary adenylate cyclase-activating polypeptide following acute spinal cord injury in rats

作　　者：徐锡金[1] 刘晓瑜[1] 梁晓[1] 霍霞[1] 

机构地区：[1]汕头大学医学院中心实验室,广东省汕头市515031

出　　处：《中国组织工程研究与临床康复》2007年第36期7195-7198,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省自然科学基金资助项目(04020247)~~

摘　　要：目的:观察大鼠脊髓损伤后垂体腺苷酸环化酶激活肽表达的变化,分析其在脊髓损伤中的意义,为进一步探索脊髓损伤后的再生修复机制和神经营养因子治疗脊髓损伤提供实验依据。方法:实验于2005-10/2006-09在汕头大学医学院生殖医学研究中心完成。①实验分组:选择清洁级健康成年雌性SD大鼠66只,随机抽签法分为损伤后1,3,7,14,28d组以及正常对照组,每组11只。②实验方法:改良的NYU装置制备脊髓损伤模型,对照组不做手术。损伤后1,3,7,14,28d麻醉取材。逆转录-聚合酶链反应扩增垂体腺苷酸环化酶激活肽。③实验评估:采用苏木精-伊红染色观察脊髓损伤后的病理变化,免疫组织化学检测阳性细胞计数,逆转录-聚合酶链反应及蛋白免疫印迹方法检测脊髓组织中垂体腺苷酸环化酶激活肽蛋白及mRNA的表达情况。结果:纳入66只大鼠,均进入结果分析。①在蛋白和mRNA水平垂体腺苷酸环化酶激活肽表达呈时间相关性,正常对照组可见垂体腺苷酸环化酶激活肽mRNA和蛋白的少量表达,脊髓损伤后1d表达开始增加,7d时表达明显增加,与3d时比较差异有显著性意义(P<0.05)。②苏木精-伊红染色结果可见脊髓灰质出血水肿坏死,神经细胞变性死亡,免疫组织化学显示正常对照组神经元中有垂体腺苷酸环化酶激活肽阳性表达,损伤后垂体腺苷酸环化酶激活肽的表达逐渐增高,损伤7d时垂体腺苷酸环化酶激活肽表达明显增加(P<0.05),至28d时表达仍较高。结论:损伤后垂体腺苷酸环化酶激活肽表达增加,内源性垂体腺苷酸环化酶激活肽的增加可能有利于受损脊髓神经元的存活与再生。AIM： To study the expression of pituitary adenylate cyclase-activating polypeptide （PACAP） after spinal cord injury （SCI） in rats and analyze its meaning in the variation so as to provide experimental evidences for further exploring the repair mechanism of SCI and treating SCI with neurotrophic factors. METHODS： The experiment was conducted in Research Center of Reproductive Medicine, Shantou University Medical College from October 2005 to September 2006. ①Sixty-six healthy female SD rats were randomly divided into five experiment groups and one control group （n =11）. ②Experiment groups were based on NYU impacting equipment to establish SCI models while control group did no operation. Rats were killed under anesthesia at 1, 3, 7, 14, 28 days after injury. PACAP was amplified using reverse transcdption-polymerase chain reaction （RT-PCR）. ③The pathological changes ware observed by hematoxylin-eosin （HE） staining, the expression of PACAP was determined with immunohistochemistry, RT-PCR and wastem blot. RESULTS： All the 66 rats ware involved in the result analysis. ①PACAP mRNA and protein expressions had a time correlation. The expressions in control group ware low and there was a step-up tendency of PACAP expression after SCI, a significant increase of PACAP expression was found at 7 days after SCI compared with 3 days （P 〈 0.05）.②HE staining revealed tissue hemorrhage, edema and necrosis, neurons degeneration and death. PACAP positive cells ware expressed in neurons detected by immunohistochemistry, and the expression increased significantly at 7 days after SCI （P 〈 0.05）. CONCLUSION： The increase of PACAP expression after SCI indicates that endogenous PACAP may promote survival and regeneration of injured spinal neurons.

关 键 词：脊髓损伤 垂体腺苷酸环化酶激活肽 组织构建 

分 类 号：R651.21[医药卫生—外科学]