【摘要】：目的本研究旨在探討賴氨酰氧化酶(LOX)能否在大鼠急性脊髓損傷(spinal cord injury,SCI)修復(fù)過程中起著重要作用。方法將96只成年雌性SD大鼠分為A、B、C、D四組,A組(n=30)為β-氨基丙腈組(β-Aminopropionitrile,BAPN,一種LOX活性抑制劑),B組(n=30)為氯化鈷組(Co Cl2,一種LOX表達(dá)促進(jìn)劑),C組(n=30)為單純損傷組(injury),D組(n=6)為正常對照組(sham)。在采用Allen’s方法成功地復(fù)制大鼠急性脊髓挫傷模型的基礎(chǔ)上,設(shè)定相同打擊力度。在損傷后不同時間段[1、3、7、14和30天(d),每個時間點(diǎn)n=6]切取實(shí)驗(yàn)脊髓段。采用免疫組化(IHC)和免疫印跡(Western blot)檢測LOX在脊髓損傷組織中的表達(dá),并分析A、B、C與D組之間差異性。應(yīng)用BBB評分法評估損傷后大鼠后肢運(yùn)動的恢復(fù)狀態(tài),同時分析A、B、C組不同時間段LOX表達(dá)與肢體運(yùn)動恢復(fù)狀態(tài)的相關(guān)性。結(jié)果BBB評分結(jié)果顯示氯化鈷組(B)后肢運(yùn)動的恢復(fù)狀態(tài)分?jǐn)?shù)明顯高于β-氨基丙腈組(A)和單純損傷組(C),且A組最低。免疫組化結(jié)果顯示:在所有實(shí)驗(yàn)時間段脊髓損傷組(A、B、C組)的LOX陽性細(xì)胞數(shù)顯著高于對照組。例如在損傷后1d,A、B、C組的LOX陽性細(xì)胞數(shù)/視野分別為27.46±3.5,64.67±3.0,和41.67±2.3,而對照組的LOX陽性細(xì)胞數(shù)只有6.04±0.2。進(jìn)而,在所有實(shí)驗(yàn)時間段的LOX陽性細(xì)胞數(shù),氯化鈷組(B)高于(A、C組),而β-氨基丙腈組(A)低于單純損傷組(C)。例如在損傷后3d,單純損傷組(C)的LOX陽性細(xì)胞數(shù)升達(dá)頂峰46.34±3.4,對應(yīng)之氯化鈷組(B)和β-氨基丙腈組(A)陽性值分別為74.81±4.9和37.64±3.6。免疫印跡結(jié)果顯示,與對照組(D)相比,氯化鈷組(B)在損傷后1d LOX表達(dá)增加,3d達(dá)到高峰,7d時有所下降,14d和30d時明顯減少;β-氨基丙腈組(A)與氯化鈷組(B)和單純損傷組(C)相比,LOX表達(dá)曲線基本相同,但表達(dá)率卻比后者明顯低弱,氯化鈷組(B)LOX表達(dá)明顯高于β-氨基丙腈組(A)和單純損傷組(C)且β-氨基丙腈組(A)最低。相同時間段間兩兩比較都具有統(tǒng)計學(xué)意義(P0.05)。結(jié)論SCI放大脊髓LOX表達(dá)。氯化鈷強(qiáng)化LOX表達(dá)加速脊髓損傷的愈合;而BAPN滅活LOX活性延緩損傷脊髓的恢復(fù)。BAPN和Co Cl2引起LOX表達(dá)的改變與BBB大鼠后肢運(yùn)動的恢復(fù)評分高低呈正相關(guān),由此說明LOX作為重要的抗損傷因子促進(jìn)急性脊髓損傷的修復(fù)。
[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 鹵3.5 鹵64.67 鹵3.0 and 41.67 鹵2.3 in group C and 6.04 鹵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 鹵3.4, and the corresponding (A) positive values of Cobalt chloride group and 尾 -aminopropionitrile group were 74.81 鹵4.9 and 37.64 鹵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.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R651.2
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本文編號：2400433