TGFβ1和GDNF在脊髓全横断损伤后大鼠的表达

发布时间：2019-05-12 12:54

【摘要】：目的:探讨成年大鼠脊髓全横断损伤(SCT)后不同时相损伤部位两端脊髓内TGFβ1和GDNF的表达变化。方法:成年SD大鼠50只,随机分5组:正常对照组、脊髓全横断损伤(于T_9-T_(10)完全横断脊髓)3d组、7d组、14d组和21d组,每组各10只动物。术后1d,3d,7d,14d及21d作BBB评分及SEP检测。每组取5只动物的脊髓,用TGFβ1和GDNF抗体行免疫组化SP法染色,另5只做Western blotting分析。在光镜下观察各组脊髓内TGFβ1和GDNF的分布及染色程度。Western blotting染色带用生物凝胶成像系统成像及Synaptic Gene Tool软件进行分析,测定阳性染色区带的光密度值。采用单因素的方差分析及LSD—q检验进行显著性检验。结果:双下肢运动功能的BBB评分在术后为0,21d仅为1.17±0.11。SEP在损伤各组均未检测到。TGFβ1表达于正常脊髓前角运动神经元的胞浆及白质内的神经胶质细胞。损伤脊髓头端,前角神经元内TGFβ1免疫反应在损伤7d开始增强,21d达最高;损伤脊髓尾端,灰质神经元TGFβ1免疫反应损伤3d开始增强,21d达最高;两端脊髓白质内神经胶质细胞的免疫反应均在14d开始增强,21d达最高。Western blotting结果显示,各损伤组头端脊髓TGFβ1蛋白含量较正常时明显减少,至损伤21d,TGFβ1的蛋白含量有所回升,但仍未达正常水平;尾端脊髓TGFβ1含量损伤后无明显变化,直到损伤21d,TGFβ1含量才明显增高。GDNF主要表达于正常脊髓的前角运动神经元,中间带神经元,后角感觉神经元及白质内的神经胶质细胞;神经元内,免疫阳性反应产物定位于胞浆及胞核。损伤脊髓头端,神经元及白质内神经胶质细胞的GDNF免疫阳性反应于损伤3d和7d明显增强,14d及21d则较前稍有减弱,神经胶质细胞的GDNF免疫阳性反应持续性增强直到21d。损伤后,尾端脊髓灰质神经元的GDNF阳性反应,比正常时略强,到损伤21d,免疫反应强度较前减弱。神经胶质细胞的免疫反应强度在损伤14d开始增强,并持续直到21d。损伤3d始,多数前角的神经元表现为胞浆着色,胞核不着色,到21d,阳性反应物又同时
[Abstract]:Aim: to investigate the expression of TGF 尾 1 and GDNF in spinal cord of adult rats at different time points after total transection of spinal cord (SCT). Methods: fifty adult SD rats were randomly divided into 5 groups: normal control group, complete transection injury of spinal cord (3 days after complete transection of spinal cord), 7 days group, 14 days group and 21 days group, with 10 animals in each group. BBB score and SEP were measured on day 1, day 3, day 7, day 14 and day 21 after operation. In each group, the spinal cord of 5 animals were stained with Western blotting 尾 1 and GDNF antibody by SP method, and the other 5 animals were analyzed by Western blotting. The distribution and staining degree of TGF 尾 1 and GDNF in spinal cord of each group were observed under light microscope. Western blotting staining band was analyzed by biological gel imaging system and Synaptic Gene Tool software, and the optical density of positive staining zone was measured. Single factor analysis of variance and LSD-q test were used for significance test. Results: the BBB score of motor function of lower extremities was 0 on the 21st day after operation, and only 1.17 卤0.11.SEP on the 21st day. The expression of BBB 尾 1 in the cytoplasm of motor neurons in the anterior horn of normal spinal cord and glial cells in white matter was not detected in all groups. At the head end of the injured spinal cord, the immune response of TGF 尾 1 in the anterior horn neurons began to increase on the 7th day and reached the highest on the 21st day, and the TGF 尾 1 immune response of the gray matter neurons began to increase at the end of the injured spinal cord on the 21st day, and reached the highest on the 21st day. The immunoreactivity of glial cells in the white matter of the spinal cord began to increase on the 14th day and reached the highest on the 21st day. Western blotting results showed that the protein content of TGF 尾 1 in the head end of the spinal cord in each injured group was significantly lower than that in the normal group, and the protein content of TGF 尾 1 increased slightly on the 21th day after injury. However, it has not yet reached the normal level; The content of TGF 尾 1 in the caudal spinal cord did not change significantly until 21 days after injury, and it was mainly expressed in the anterior horn motoneurons, intermediate zone neurons, posterior horn sensory neurons and glial cells in the white matter of the normal spinal cord. In neurons, immunopositive products were located in cytoplasm and nucleus. The GDNF immunoreactivity of neuroglial cells in neurons and white matter of injured spinal cord increased significantly on the 3rd and 7th day after injury, and decreased slightly on the 14th and 21st day after injury. The GDNF immunoreactivity of the glial cells continued to increase until 21d. After injury, the GDNF positive reaction of gray matter neurons in the tail spinal cord was slightly stronger than that in the normal group. At the 21st day of injury, the intensity of immune response was weaker than that before injury. The immune response intensity of glial cells began to increase on the 14th day after injury and lasted until 21 days. From 3 days after injury, most of the neurons in the anterior horn showed cytoplasmic staining, and the nucleus was not stained. On the 21st day, the positive reactants were at the same time.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R361

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