摘要
目的建立大鼠脊髓损伤动物模型,分析血管内皮生长因子(VEGF)及缺氧诱导因子-1α(HIF-1α)表达与脊髓损伤的相关性及二者之间的相关性,探讨其在继发性脊髓损伤发生发展过程中的作用。方法健康成熟Wistar大鼠144只,雌雄不限,体质量180～220g。随机分为脊髓损伤组和假手术组。假手术组大鼠行椎板切除术,脊髓损伤组大鼠参照Allen法制作脊髓损伤模型。2组分别于术后1、3、6、12、24、48、72、168、336h对8只大鼠进行Tarlov评分,评分后处死。取损伤区脊髓组织进行苏木精-伊红(HE)染色观察脊髓组织病理学变化,免疫组织化学法测定VEGF及HIF-1α表达并对阳性细胞进行计数,做统计学分析和相关性分析。结果各组动物均存活至实验完成。Tarlov评分结果显示:假手术组大鼠术后各时间点评分变化不明显,脊髓损伤组大鼠术后各时间点评分均较假手术组显著降低(P<0.05),其中术后24、168 h降低最明显(1.99、2.16)。脊髓损伤组术后1～6 h,损伤局部脊髓组织结构紊乱、水肿;术后168 h内,损伤区域扩大,大量炎性细胞浸润;术后336 h损伤减轻,但仍可见较多炎性细胞浸润。免疫组织化学检测,假手术组大鼠VEGF及HIF-1α阳性细胞较少;脊髓损伤组大鼠术后6 h,VEGF及HIF-1α阳性细胞数开始增多,HIF-1α于术后24、168 h出现2次高峰,VEGF于术后168 h出现高峰,各时间点HIF-1α及VEGF阳性细胞数均较假手术组显著增加(P<0.05)。VEGF与HIF-1α表达呈正相关(r=0.526,P<0.05)。结论大鼠脊髓损伤后HIF-1α、VEGF表达升高,与继发性脊髓损伤密切相关,其表达规律与损伤时间具有相关性;HIF-1α与VEGF表达呈正相关性。
Objective To observe expressing rule of hypoxia inducible factor-1α（HIF-1α） and vascular endothelial growth factor （VEGF） in secondary spinal cord injury, and investigate the function in the secondary spinal cord injury by establishing rat spinal cord injury animal models, immunohistochemical detection of VEGF expression and HIF-1α at different time points after spinal cord injury, which provides the theory basis for clinical treatment of spinal cord injury. Methods A total of 144 healthy mature Wistar rats, unlimited gender, body weight was 180 - 220 g, provided by Laboratory Animal Center of China Medical University. The rats were randomly divided into spinal cord injury group and sham group. Rats in sham group were performed laminectomy, and rats in spinal cord injury group were established animal models of spinal cord injury accorded to Allen＇s method. Rats in both groups were scored accorded to Tarlov＇s method at 1, 3, 6, 12, 24, 48, 72, 168, 336 hours after operation, there were 8 rats in every time points. Rats were sacrificed to get spinal cord of injured regions after scoring. The pathological changes of spinal cord by hematoxylin--eosin （HE） staining, expression of HIF-1α and VEGF at different time points by immunohistochemieal detection were observed, and statistical and correlation were analyzed. Results All the rats were alive until the finish of experiment. Results of Tarlov score showed that scores of rats in sham group nearly were not changed, scores of rats in spinal cord injury group degraded significantly compared with sham group at all the time points（P 〈 0.05）, at 24-hour, 168-hour degraded most significantly（1.99, 2.16）. In spinal cord injury group, structure of spinal cord was in disorder, there were obvious edema in spinal cord from 1-hour to 6-hour after operation; In 168 -hour after operation, damaging region expanding, there were still numerous inflammatory cells infiltrated; In 336-hour after operation, the damage lessened, but there were still numerous inflammatory cells infiltrated. The immunohistochemical results showed the number of HIF-1α positive cells and VEGF positive cells were both less in sham group; the number of HIF-1α positive cells and VEGF positive cells increased at 6-hour after spinal cord injury. The 168-hour was at peak of VEGF and 24, 168 hours were at peak of HIF-lct（P 〈 0.05）. The expression of VEGF and HIF-1α was positively correlated （r = 0.526, P 〈 0.05）. Conclusion It is demonstrated that expression of VEGF and HIF-1α are increased after spinal cord injury, and their expression law are related to the damage time, the expression of HIF-1α and VEGF is positively correlated in secondary spinal cord.
出处
《生物医学工程与临床》
CAS
2013年第2期178-183,共6页
Biomedical Engineering and Clinical Medicine
