堿性成纖維細(xì)胞生長因子對大鼠脊髓缺血再灌注損傷后的保護(hù)作用
發(fā)布時間：2018-08-29 18:29
【摘要】： 目的觀察應(yīng)用外源性堿性成纖維細(xì)胞生長因子(basic fibroblast growth factor,bFGF)對脊髓缺血再灌注損傷后的作用。 方法72只健康雄性成年大鼠,隨機(jī)分為正常對照組(n=24),生理鹽水組(n=24)和bFGF治療組(n=24)。通過阻斷腹主動脈建立脊髓缺血再灌注損傷模型,于損傷平面經(jīng)蛛網(wǎng)膜下腔置細(xì)導(dǎo)管,bFGF治療組分別于恢復(fù)血流前10min,恢復(fù)血供后6h、12h、24h、48h經(jīng)細(xì)導(dǎo)管注入bFGF,10mg/kg。生理鹽水組在相同時間注入等量生理鹽水,然后不同時間處死動物取材(n=4)。光學(xué)顯微鏡觀察脊髓組織病理變化;硫代巴比妥酸法測定血漿和脊髓組織丙二醛含量;全自動生化分析儀測定血漿肌酸磷酸激酶、谷草轉(zhuǎn)氨酶和乳酸脫氫酶含量;放射免疫法測定脊髓組織勻漿內(nèi)皮素水平;差速離心法測定細(xì)胞線粒體鈣含量;取部分脊髓測定組織濕/干重比值;TUNEL法檢測細(xì)胞凋亡;免疫組化檢測脊髓組織p53表達(dá)的變化情況。 結(jié)果(1)光鏡觀察:和bFGF治療組相比較,生理鹽水組脊髓神經(jīng)元明顯退變,減少、溶解或壞死。尼氏體變淺或消失。脊髓灰質(zhì)出現(xiàn)片狀出血灶,大量膠質(zhì)細(xì)胞浸潤;(2)恢復(fù)血流灌注1h,生理鹽水組與正常對照組比較,血漿和脊髓的各項生化指標(biāo)顯著增高(P＜0.05);使用bFGF后,血漿及脊髓各項測定指標(biāo)較生理鹽水組比較明顯降低(P＜0.05)。(3)TUNEL染色顯示:bFGF治療組24h、48h、72h、96h時相點凋亡陽性細(xì)胞明顯低于生理鹽水組(P＜0.05)。(4)免疫組化染色顯示:bFGF治療組24h、48h、72h、96h時相點p53的陽性細(xì)胞數(shù)明顯低于生理鹽水組(P＜0.05)。 結(jié)論堿性成纖維細(xì)胞生長因子明顯減輕脊髓缺血再灌注造成的組織損傷,對損傷脊髓有保護(hù)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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R363
本文編號：2212047
[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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R363
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