發(fā)布時間：2019-01-18 20:21

【摘要】：采用免疫組織化學方法檢測脊髓損傷(spinal cord injury,SCI)后各組不同時段生長相關蛋白-43(Growth associated protein-43,GAP-43)和熱休克蛋白27(Heat shock protein27,HSP27)表達情況;各組大鼠在存活期間作后肢Basso Beattie Bresnahan(BBB)運動功能評分;用蘇木精-伊紅(hematoxylin-eosin,HE)染色法觀察損傷的脊髓組織各組不同時段病理變化。以確定甲潑尼龍琥珀酸鈉(Methylpredniolone Sodium Succinate,MPss)鞘內注射后是否參與了脊髓神經元內GAP-43和HSP27表達的調節(jié),進一步闡明MPss鞘內注射對急性SCI修復中的可能作用機制。健康成年大鼠體重(210±10)g雌性大鼠90只,隨機分為治療組、觀察組、對照組,每組30只。治療組予以脊髓損傷后MPss 3 mg/kg溶解于0.1 mL 0.9%氯化鈉中,鞘內注射;觀察組予以單純手術暴露脊髓,不損傷脊髓;對照組予以脊髓損傷后0.9%氯化鈉0.1 m L,鞘內注射。于術后第1,3,7,14,28天進行組織學觀察分析、行為學觀察。組織學結果:顯示GAP-43在大鼠脊髓損傷后均表達增強,觀察組有少量表達。治療組、對照組均出現(xiàn)高表達,在脊髓損傷后1,3,7天表達強度逐漸上升,在第7天達到最高,后表達逐漸下降,對照組第14天降至接近對照組,第28天治療組、對照組表達均降至相同水平,但仍稍高于觀察組。治療組在早期表達數(shù)量及表達強度均高于對照組,隨著時間延長,降至與觀察組相同水平。圖像分析結果顯示,治療組脊髓前角的陽性反應在1,3,7,14d點高于對照組,3d差異最明顯,差異有顯著性(P0.05)。HSP27在對照組呈低水平表達,治療組及觀察組均在1d達到高峰,治療組在1,3d的陽性細胞數(shù)及陽性染色強度均強于對照組,后表達逐漸下降,治療組和對照組第7天降至接近觀察。治療組脊髓灰質陽性細胞較對照組高,最明顯差別在術后1d,差異有統(tǒng)計學意義(P0.01)。HE染色結果:治療組、對照組均可見脊髓組織內細胞水腫、結構紊亂、淋巴細胞浸潤尼氏體融解等病理變化,治療組病理改變輕于觀察組。觀察組脊髓結構正常,無組織內細胞水腫、結構紊亂、淋巴細胞浸潤尼氏體融解等病理變化。脊髓組織HE染色結果表明,甲潑尼龍琥珀酸鈉鞘內注射可減輕脊髓損傷后的病理改變結果行為學觀察:BBB評分提示甲潑尼龍琥珀酸鈉鞘內注射可改善大鼠脊髓損傷后后肢運動功能,術后第7d,14d,28d,治療組、對照組間差異有統(tǒng)計學意義(P0.05),治療組較對照組恢復明顯。甲潑尼龍琥珀酸鈉鞘內注射可以促進損傷脊髓神經細胞修復,其作用機制可能與甲潑尼龍琥珀酸鈉鞘內注射增強損傷脊髓神經細胞GAP-43和HSP27的表達有關。
[Abstract]:The expression of growth-associated protein-43 (Growth associated protein-43,GAP-43) and heat shock protein (27 (Heat shock protein27,HSP27) were detected by immunohistochemical method after spinal cord injury (spinal cord injury,SCI). The Basso Beattie Bresnahan (BBB) motor function score of hind limbs and the pathological changes of injured spinal cord tissues were observed by hematoxylin-eosin (hematoxylin-eosin,HE) staining. To determine whether intrathecal methylprednisolone sodium succinate (Methylpredniolone Sodium Succinate,MPss) involved in the regulation of GAP-43 and HSP27 expression in spinal cord neurons, and to elucidate the possible mechanism of intrathecal MPss injection in the repair of acute SCI. 90 healthy adult rats weighing (210 鹵10) g female rats were randomly divided into treatment group, observation group and control group with 30 rats in each group. After spinal cord injury, MPss 3 mg/kg was dissolved in 0. 1 mL 0.9% sodium chloride and injected intrathecally in the treatment group. The control group was given 0.9% sodium chloride 0.1 mL intrathecal injection after spinal cord injury. Histologic observation and behavioral observation were performed on the 1st day, 3rd day and 1428 th day after operation. Histological results: the expression of GAP-43 was increased after spinal cord injury in rats, and a little in observation group. In the treatment group and the control group, the expression level increased gradually on the 7th day after spinal cord injury, reached the highest level on the 7th day, and then decreased gradually, the control group decreased to close to the control group on the 14th day, and the treatment group on the 28th day. The expression of the control group decreased to the same level, but still slightly higher than the observation group. The quantity and intensity of expression in the treatment group were higher than those in the control group, and decreased to the same level as the observation group. The results of image analysis showed that the positive reaction of spinal cord anterior horn in the treatment group was higher than that in the control group at 714 days after treatment, and the difference was most significant on the 3rd day (P0.05). The expression of HSP27 in the control group was lower than that in the control group. The number of positive cells and the intensity of positive staining in the treatment group were stronger than those in the control group on the 1st day, then the expression decreased gradually, and decreased to close observation on the 7th day in the treatment group and the control group. The positive cells of spinal cord in the treatment group were higher than those in the control group, the most obvious difference was on the 1st day after operation. The difference was statistically significant (P0.01). HE staining result: in the treatment group and the control group, the cell edema and structural disorder in the spinal cord tissue were observed. The pathological changes of lymphocytic infiltration and Nissl body fusion were lighter in the treatment group than in the observation group. In the observation group, the structure of spinal cord was normal, no cell edema, structural disorder, lymphocyte infiltration and melting of Nissl body, and other pathological changes. HE staining of spinal cord tissue showed that, Intrathecal injection of methylprednisolone sodium succinate alleviated the pathological changes after spinal cord injury. BBB score indicated that intrathecal injection of methylprednisolone succinate sodium could improve the motor function of hind limbs in rats with spinal cord injury. The difference between the treatment group and the control group was statistically significant (P0.05), the treatment group was more obvious than the control group. Intrathecal injection of methylprednisolone sodium succinate can promote the repair of injured spinal cord neurons, and its mechanism may be related to the enhancement of the expression of GAP-43 and HSP27 in injured spinal cord neurons by intrathecal injection of methylprednisolone sodium succinate.
【學位授予單位】：河北北方學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R651.2

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本文編號：2411079