【摘要】：目的:探討成年大鼠脊髓全橫斷損傷(SCT)后不同時相損傷部位兩端脊髓內(nèi)TGFβ1和GDNF的表達(dá)變化。 方法:成年SD大鼠50只,隨機(jī)分5組:正常對照組、脊髓全橫斷損傷(于T_9-T_(10)完全橫斷脊髓)3d組、7d組、14d組和21d組,每組各10只動物。術(shù)后1d,3d,7d,14d及21d作BBB評分及SEP檢測。每組取5只動物的脊髓,用TGFβ1和GDNF抗體行免疫組化SP法染色,另5只做Western blotting分析。在光鏡下觀察各組脊髓內(nèi)TGFβ1和GDNF的分布及染色程度。Western blotting染色帶用生物凝膠成像系統(tǒng)成像及Synaptic Gene Tool軟件進(jìn)行分析,測定陽性染色區(qū)帶的光密度值。采用單因素的方差分析及LSD—q檢驗(yàn)進(jìn)行顯著性檢驗(yàn)。 結(jié)果:雙下肢運(yùn)動功能的BBB評分在術(shù)后為0,21d僅為1.17±0.11。SEP在損傷各組均未檢測到。TGFβ1表達(dá)于正常脊髓前角運(yùn)動神經(jīng)元的胞漿及白質(zhì)內(nèi)的神經(jīng)膠質(zhì)細(xì)胞。損傷脊髓頭端,前角神經(jīng)元內(nèi)TGFβ1免疫反應(yīng)在損傷7d開始增強(qiáng),21d達(dá)最高;損傷脊髓尾端,灰質(zhì)神經(jīng)元TGFβ1免疫反應(yīng)損傷3d開始增強(qiáng),21d達(dá)最高;兩端脊髓白質(zhì)內(nèi)神經(jīng)膠質(zhì)細(xì)胞的免疫反應(yīng)均在14d開始增強(qiáng),21d達(dá)最高。Western blotting結(jié)果顯示,各損傷組頭端脊髓TGFβ1蛋白含量較正常時明顯減少,至損傷21d,TGFβ1的蛋白含量有所回升,但仍未達(dá)正常水平;尾端脊髓TGFβ1含量損傷后無明顯變化,直到損傷21d,TGFβ1含量才明顯增高。GDNF主要表達(dá)于正常脊髓的前角運(yùn)動神經(jīng)元,中間帶神經(jīng)元,后角感覺神經(jīng)元及白質(zhì)內(nèi)的神經(jīng)膠質(zhì)細(xì)胞;神經(jīng)元內(nèi),免疫陽性反應(yīng)產(chǎn)物定位于胞漿及胞核。損傷脊髓頭端,神經(jīng)元及白質(zhì)內(nèi)神經(jīng)膠質(zhì)細(xì)胞的GDNF免疫陽性反應(yīng)于損傷3d和7d明顯增強(qiáng),14d及21d則較前稍有減弱,神經(jīng)膠質(zhì)細(xì)胞的GDNF免疫陽性反應(yīng)持續(xù)性增強(qiáng)直到21d。損傷后,尾端脊髓灰質(zhì)神經(jīng)元的GDNF陽性反應(yīng),比正常時略強(qiáng),到損傷21d,免疫反應(yīng)強(qiáng)度較前減弱。神經(jīng)膠質(zhì)細(xì)胞的免疫反應(yīng)強(qiáng)度在損傷14d開始增強(qiáng),并持續(xù)直到21d。損傷3d始,多數(shù)前角的神經(jīng)元表現(xiàn)為胞漿著色,胞核不著色,到21d,陽性反應(yīng)物又同時
[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 鹵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.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R361

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