碱性成纤维细胞生长因子对大鼠脊髓缺血再灌注损伤后的保护作用

发布时间：2018-08-29 18:29

【摘要】： 目的观察应用外源性碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对脊髓缺血再灌注损伤后的作用。方法72只健康雄性成年大鼠,随机分为正常对照组(n=24),生理盐水组(n=24)和bFGF治疗组(n=24)。通过阻断腹主动脉建立脊髓缺血再灌注损伤模型,于损伤平面经蛛网膜下腔置细导管,bFGF治疗组分别于恢复血流前10min,恢复血供后6h、12h、24h、48h经细导管注入bFGF,10mg/kg。生理盐水组在相同时间注入等量生理盐水,然后不同时间处死动物取材(n=4)。光学显微镜观察脊髓组织病理变化;硫代巴比妥酸法测定血浆和脊髓组织丙二醛含量;全自动生化分析仪测定血浆肌酸磷酸激酶、谷草转氨酶和乳酸脱氢酶含量;放射免疫法测定脊髓组织匀浆内皮素水平;差速离心法测定细胞线粒体钙含量;取部分脊髓测定组织湿/干重比值;TUNEL法检测细胞凋亡;免疫组化检测脊髓组织p53表达的变化情况。结果(1)光镜观察:和bFGF治疗组相比较,生理盐水组脊髓神经元明显退变,减少、溶解或坏死。尼氏体变浅或消失。脊髓灰质出现片状出血灶,大量胶质细胞浸润;(2)恢复血流灌注1h,生理盐水组与正常对照组比较,血浆和脊髓的各项生化指标显著增高(P＜0.05);使用bFGF后,血浆及脊髓各项测定指标较生理盐水组比较明显降低(P＜0.05)。(3)TUNEL染色显示:bFGF治疗组24h、48h、72h、96h时相点凋亡阳性细胞明显低于生理盐水组(P＜0.05)。(4)免疫组化染色显示:bFGF治疗组24h、48h、72h、96h时相点p53的阳性细胞数明显低于生理盐水组(P＜0.05)。结论碱性成纤维细胞生长因子明显减轻脊髓缺血再灌注造成的组织损伤,对损伤脊髓有保护作用。
[Abstract]:Objective to observe the effect of exogenous basic fibroblast growth factor (basic fibroblast growth factor,bFGF) on spinal cord ischemia reperfusion injury. Methods Seventy-two healthy male adult rats were randomly divided into normal control group (n = 24), saline group (n = 24) and bFGF group (n = 24). The spinal cord ischemia-reperfusion injury model was established by blocking abdominal aorta. In the injury plane treated group treated with subarachnoid catheterization, 10 minutes before blood flow was restored, and 6 hours after restoration of blood supply, 24 hours and 48 hours after blood supply were restored, bFGF,10mg/kg. was injected through the catheter. Saline group was injected with the same amount of saline at the same time, and then animals were sacrificed at different time points (nun4). The content of malondialdehyde (MDA) in plasma and spinal cord was determined by thiobarbituric acid method, and the contents of creatine phosphokinase, glutamic oxaloacetic transaminase and lactate dehydrogenase were measured by automatic biochemical analyzer. The levels of endothelin in spinal cord homogenate were measured by radioimmunoassay, mitochondrial calcium content was measured by differential centrifugation, and apoptosis was detected by Tunel method. The expression of p53 in spinal cord was detected by immunohistochemistry. Results (1) Light microscopic observation: compared with the bFGF group, the spinal cord neurons in the saline group were obviously degenerative, decreased, dissolved or necrotized. The Nissl body becomes shallower or disappears. (2) after 1 hour of blood perfusion, the biochemical indexes of plasma and spinal cord in the saline group were significantly higher than those in the normal control group (P < 0. 05). Compared with the normal saline group, the plasma and spinal cord indexes were significantly decreased (P < 0. 05). (3) TUNEL staining showed that the apoptotic positive cells in the treatment group were significantly lower than those in the saline group at 24 h, 48 h, 72 h and 96 h (P < 0. 05). (4). The immunohistochemical staining showed that the percentage of apoptotic cells in the treatment group was significantly lower than that in the control group at 24 h, 48 h, 72 h, 96 h. The number of p53 positive cells at phase point was significantly lower than that in saline group (P < 0.05). Conclusion basic fibroblast growth factor can attenuate the injury of spinal cord induced by ischemia reperfusion and has protective effect on spinal cord injury.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363

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