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目的:观察银杏叶提取物(EGb)对大鼠实验性脊髓损伤后组织结构及运动功能恢复的作用,探讨其对急性脊髓损伤的作用机制。方法:120只SD雄性大鼠,随机分为假手术组(A组)、损伤对照组(B组)、甲基强的松龙(MP)治疗组(C组)和EGb治疗组(D组),每组30只。B、C、D组用Allen′s法以50g·cm致伤大鼠T9脊髓制作损伤模型,B组为单纯脊髓损伤,不给药;C组为脊髓损伤后30min内,由腹腔注入MP30mg/kg;D组为术后至处死前每天腹腔给予EGb17.5mg/kg;A组只打开T9椎板,不打击脊髓,不给药。术后24h、3d、5d、7d、14d对大鼠进行脊髓运动功能(BBB)评分。于术后24h、3d、5d、7d、14d处死动物(n=6),取T9节段脊髓,切片苏木素-伊红(HE)染色观察脊髓大体组织结构变化,用免疫组织化学方法检测B细胞淋巴瘤/白血病基因-2(Bcl-2)和B细胞淋巴瘤/白血病基因伴随蛋白x(Bax)在脊髓前角运动神经元中的表达变化情况。结果:各时间点B、C、D组大鼠脊髓运动功能(BBB)评分均显著低于A组(P<0.01),伤后7d、14d时C、D组评分显著高于B组(P<0.05),各时间点C组与D组评分比较无统计学意义(P>0.05)。HE染色C组和D组大鼠脊髓损伤区较B组坏死程度轻、形成囊腔少,A组正常;1周后D组较C组片状出血灶少,神经细胞肿胀不明显。各时间点B、C、D组大鼠损伤脊髓前角运动神经元中Bcl-2阳性细胞数均显著高于A组(P<0.01),C、D组显著高于B组(P<0.01),7d、14d时D组显著高于C组(P<0.05);各时间点B、C、D组大鼠损伤脊髓前角运动神经元中Bax阳性细胞数均显著高于A组(P<0.01),C、D组显著低于B组(P<0.01),7d、14d时D组显著低于C组(P<0.01)。结论:EGb可能通过抑制Bax表达、提高Bcl-2表达,抑制脊髓损伤后神经元凋亡,在运动功能恢复、损伤脊髓组织保护上发挥其有益作用,1周后EGb仍能抑制脊髓损害后的继发性损伤。
Objective: To observe the effect of Ginkgo biloba extract (EGb) on the recovery of tissue structure and motor function after experimental spinal cord injury in rats, and to explore its mechanism of action on acute spinal cord injury. METHODS: One hundred and twenty SD male rats were randomly divided into sham operation group (group A), injury control group (group B), methylprednisolone (MP) treatment group (group C) and EGb treatment group (group D). ), 30 per group. In group B, C, and D, Allen’s method was used to infuse the rat spinal cord T9 at 50g·cm to make an injury model. In group B, the spinal cord was injured and not administered; in group C, the MP30mg was injected into the abdominal cavity within 30min after spinal cord injury. Kg; D group was given EGb 17.5mg/kg intraperitoneally before to death; in group A, only T9 lamina was opened, no spinal cord was struck, and no administration was given. Rats were evaluated for spinal motor function (BBB) at 24h, 3d, 5d, 7d, and 14d after operation. Animals were sacrificed 24h, 3d, 5d, 7d, and 14d after surgery (n=6). T9 spinal cord was taken and sections of hematoxylin-eosin (HE) were stained to observe the changes in the structure of spinal cord. B-cells were detected by immunohistochemistry. Changes of Expression of Lymphoma/Leukemia Gene-2 (Bcl-2) and B-cell Lymphoma/Leukemia Gene Companion Protein x (Bax) in Anterior Spinal Cord Motoneuron of Spinal Cord. RESULTS: The scores of spinal motor function (BBB) in rats of groups B, C, and D at each time point were significantly lower than those in group A (P<0.01). Scores in group C and D at days 7 and 14 after injury were significantly higher than those in group B (P<0.01). <0.05), There was no significant difference between the C group and the D group at each time point (P>0.05). Compared with group B, the necrotic area of rats in group C and group D was less than that in group B. The formation of cyst was less in group A. Normal group A was normal; after one week, group D had fewer flaky hemorrhages than group C, and nerve cell swelling was not obvious. At each time point, the number of Bcl-2 positive cells in the injured spinal cord motoneurons in the B, C, and D groups was significantly higher than that in the A group (P<0.01), and significantly higher in the C and D groups than in the B group (P<0.01). At 7d and 14d, the D group was significantly higher than that of the C group (P<0.05); at each time point, the number of Bax positive cells in the injured spinal cord motoneurons of the B, C, and D groups was significantly higher than that of the A group (P <0.01), C, D group was significantly lower than B group (P <0.01), 7d, 14d D group was significantly lower than C group (P <0.01). CONCLUSION: EGb may inhibit the expression of Bax, increase the expression of Bcl-2, and inhibit neuronal apoptosis after spinal cord injury. It plays a beneficial role in recovery of motor function and protection of injured spinal cord tissue. EGb can still inhibit spinal cord injury after 1 week. Secondary injury.