赖氨酰氧化酶在大鼠急性脊髓损伤修复过程中的作用
发布时间：2019-01-04 14:53
【摘要】：目的本研究旨在探讨赖氨酰氧化酶(LOX)能否在大鼠急性脊髓损伤(spinal cord injury,SCI)修复过程中起着重要作用。方法将96只成年雌性SD大鼠分为A、B、C、D四组,A组(n=30)为β-氨基丙腈组(β-Aminopropionitrile,BAPN,一种LOX活性抑制剂),B组(n=30)为氯化钴组(Co Cl2,一种LOX表达促进剂),C组(n=30)为单纯损伤组(injury),D组(n=6)为正常对照组(sham)。在采用Allen’s方法成功地复制大鼠急性脊髓挫伤模型的基础上,设定相同打击力度。在损伤后不同时间段[1、3、7、14和30天(d),每个时间点n=6]切取实验脊髓段。采用免疫组化(IHC)和免疫印迹(Western blot)检测LOX在脊髓损伤组织中的表达,并分析A、B、C与D组之间差异性。应用BBB评分法评估损伤后大鼠后肢运动的恢复状态,同时分析A、B、C组不同时间段LOX表达与肢体运动恢复状态的相关性。结果BBB评分结果显示氯化钴组(B)后肢运动的恢复状态分数明显高于β-氨基丙腈组(A)和单纯损伤组(C),且A组最低。免疫组化结果显示:在所有实验时间段脊髓损伤组(A、B、C组)的LOX阳性细胞数显著高于对照组。例如在损伤后1d,A、B、C组的LOX阳性细胞数/视野分别为27.46±3.5,64.67±3.0,和41.67±2.3,而对照组的LOX阳性细胞数只有6.04±0.2。进而,在所有实验时间段的LOX阳性细胞数,氯化钴组(B)高于(A、C组),而β-氨基丙腈组(A)低于单纯损伤组(C)。例如在损伤后3d,单纯损伤组(C)的LOX阳性细胞数升达顶峰46.34±3.4,对应之氯化钴组(B)和β-氨基丙腈组(A)阳性值分别为74.81±4.9和37.64±3.6。免疫印迹结果显示,与对照组(D)相比,氯化钴组(B)在损伤后1d LOX表达增加,3d达到高峰,7d时有所下降,14d和30d时明显减少;β-氨基丙腈组(A)与氯化钴组(B)和单纯损伤组(C)相比,LOX表达曲线基本相同,但表达率却比后者明显低弱,氯化钴组(B)LOX表达明显高于β-氨基丙腈组(A)和单纯损伤组(C)且β-氨基丙腈组(A)最低。相同时间段间两两比较都具有统计学意义(P0.05)。结论SCI放大脊髓LOX表达。氯化钴强化LOX表达加速脊髓损伤的愈合;而BAPN灭活LOX活性延缓损伤脊髓的恢复。BAPN和Co Cl2引起LOX表达的改变与BBB大鼠后肢运动的恢复评分高低呈正相关,由此说明LOX作为重要的抗损伤因子促进急性脊髓损伤的修复。
[Abstract]:Objective to investigate whether lysyl oxidase (LOX) plays an important role in the repair of acute spinal cord injury (spinal cord injury,SCI) in rats. Methods 96 adult female SD rats were divided into four groups: group A (n = 30) was 尾 -aminopropionitrile group (n = 30), LOX activity inhibitor), B group (n = 30) was cobalt chloride group (Co Cl2,), group A (n = 30) was 尾 -aminopropionitrile group (n = 30). A kind of LOX expression enhancer,), C group (nong30), is (injury), D group with simple injury (nong6) and normal control group (sham). On the basis of successful replication of acute spinal cord contusion rat model by Allen's method, the same strike intensity was set up. The experimental spinal cord segments were removed at different time points of (d), at different time points after injury. Immunohistochemical (IHC) and Western blot (Western blot) were used to detect the expression of LOX in spinal cord injury tissues. BBB score was used to evaluate the recovery state of hindlimb movement in rats after injury, and the correlation between LOX expression and limb motor recovery was analyzed in group A BU C at different time points. Results BBB score showed that the recovery state score of (B) hind limb movement in Cobalt chloride group was significantly higher than that in 尾 -aminopropionitrile group (A) and simple injury group (C),) and the lowest in group A. Immunohistochemical results showed that the number of LOX positive cells in the spinal cord injury group was significantly higher than that in the control group. For example, the number of LOX positive cells / visual field were 27.46 卤3.5 卤64.67 卤3.0 and 41.67 卤2.3 in group C and 6.04 卤0.2 in control group on the 1st day after injury. Furthermore, the number of LOX positive cells in the cobalt chloride group was higher than that in the control group (APC group), while the (A) in 尾 -aminopropionitrile group was lower than that in the simple injury group. For example, on the 3rd day after injury, the number of LOX positive cells in (C) increased to a peak of 46.34 卤3.4, and the corresponding (A) positive values of Cobalt chloride group and 尾 -aminopropionitrile group were 74.81 卤4.9 and 37.64 卤3.6, respectively. The results of Western blot showed that compared with the control group, the expression of (B) in Cobalt chloride group increased at 1 day after injury, reached the peak at 3 days, decreased at 7 days, and decreased at 14 and 30 days. The expression curve of LOX in 尾 -aminopropionitrile group was similar to that in cobalt chloride group (B) and simple damaged group (C), but the expression rate of LOX in 尾 -aminopropionitrile group was significantly lower than that in the latter group. The expression of (B) LOX in cobalt chloride group was significantly higher than that in 尾 -aminopropionitrile group (A) and simple injury group (C) and the lowest in 尾 -aminopropionitrile group (A). In the same time period, the comparison was statistically significant (P0.05). Conclusion SCI amplifies the expression of LOX in spinal cord. Cobalt chloride enhanced LOX expression to accelerate the healing of spinal cord injury. The activity of BAPN inactivated LOX delayed the recovery of injured spinal cord. The changes of LOX expression induced by BAPN and Co Cl2 were positively correlated with the recovery score of hindlimb movement in BBB rats, which indicated that LOX as an important anti-injury factor promoted the repair of acute spinal cord injury.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
，
本文编号：2400433
[Abstract]:Objective to investigate whether lysyl oxidase (LOX) plays an important role in the repair of acute spinal cord injury (spinal cord injury,SCI) in rats. Methods 96 adult female SD rats were divided into four groups: group A (n = 30) was 尾 -aminopropionitrile group (n = 30), LOX activity inhibitor), B group (n = 30) was cobalt chloride group (Co Cl2,), group A (n = 30) was 尾 -aminopropionitrile group (n = 30). A kind of LOX expression enhancer,), C group (nong30), is (injury), D group with simple injury (nong6) and normal control group (sham). On the basis of successful replication of acute spinal cord contusion rat model by Allen's method, the same strike intensity was set up. The experimental spinal cord segments were removed at different time points of (d), at different time points after injury. Immunohistochemical (IHC) and Western blot (Western blot) were used to detect the expression of LOX in spinal cord injury tissues. BBB score was used to evaluate the recovery state of hindlimb movement in rats after injury, and the correlation between LOX expression and limb motor recovery was analyzed in group A BU C at different time points. Results BBB score showed that the recovery state score of (B) hind limb movement in Cobalt chloride group was significantly higher than that in 尾 -aminopropionitrile group (A) and simple injury group (C),) and the lowest in group A. Immunohistochemical results showed that the number of LOX positive cells in the spinal cord injury group was significantly higher than that in the control group. For example, the number of LOX positive cells / visual field were 27.46 卤3.5 卤64.67 卤3.0 and 41.67 卤2.3 in group C and 6.04 卤0.2 in control group on the 1st day after injury. Furthermore, the number of LOX positive cells in the cobalt chloride group was higher than that in the control group (APC group), while the (A) in 尾 -aminopropionitrile group was lower than that in the simple injury group. For example, on the 3rd day after injury, the number of LOX positive cells in (C) increased to a peak of 46.34 卤3.4, and the corresponding (A) positive values of Cobalt chloride group and 尾 -aminopropionitrile group were 74.81 卤4.9 and 37.64 卤3.6, respectively. The results of Western blot showed that compared with the control group, the expression of (B) in Cobalt chloride group increased at 1 day after injury, reached the peak at 3 days, decreased at 7 days, and decreased at 14 and 30 days. The expression curve of LOX in 尾 -aminopropionitrile group was similar to that in cobalt chloride group (B) and simple damaged group (C), but the expression rate of LOX in 尾 -aminopropionitrile group was significantly lower than that in the latter group. The expression of (B) LOX in cobalt chloride group was significantly higher than that in 尾 -aminopropionitrile group (A) and simple injury group (C) and the lowest in 尾 -aminopropionitrile group (A). In the same time period, the comparison was statistically significant (P0.05). Conclusion SCI amplifies the expression of LOX in spinal cord. Cobalt chloride enhanced LOX expression to accelerate the healing of spinal cord injury. The activity of BAPN inactivated LOX delayed the recovery of injured spinal cord. The changes of LOX expression induced by BAPN and Co Cl2 were positively correlated with the recovery score of hindlimb movement in BBB rats, which indicated that LOX as an important anti-injury factor promoted the repair of acute spinal cord injury.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
，
本文编号：2400433
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