胞二磷胆碱结合康复训练对脑缺血大鼠行为学及神经可塑性的影响
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摘要
研究目的
研究胞二磷胆碱结合康复训练对脑缺血大鼠运动功能恢复及对钙结合蛋白、β微管蛋白表达的影响,为临床药物结合康复训练治疗脑血管病提供理论依据。
研究方法
按随机数字表法,将120只成年雄性SD大鼠采用Longa线栓法制作大鼠左侧大脑中动脉闭塞(MCAO)模型,选出造模成功后符合标准的96只大鼠随机分为对照组、胞二磷胆碱组、康复训练组、胞二磷胆碱结合康复训练组4组,每组24只。造模成功后3d,胞二磷胆碱组开始给予胞二磷胆碱(每天500mg/kg体重),康复训练组开始进行滚笼、平衡木、转棒、网屏等训练,胞二磷胆碱结合康复训练组既给予胞二磷胆碱,又给予康复训练,对照组不做任何干预。各组分别在造模后7、14、21d时进行行为学评估综合评分,同时在相应时间点,经灌流固定后断头取脑；各组取脑组织行免疫组化法观察缺血周围皮质S-100.β-tubulin的表达情况。采用SPSS13.0统计学软件进行单因素方差分析以及Newman-Keuls检验,P<0.05认为差异有统计学意义。
结果
1.大脑中动脉梗塞成功造模后各组大鼠均不同程度出现神经功能缺损的体征。
2.神经行为学评估：
各组大鼠随时间推移行为学评分均逐渐减小,胞二磷胆碱组、康复训练组、胞二磷胆碱结合康复训练组造模后7d行为学评分分别与同一时间点对照组比较,差异无统计学意义(P>0.05)；胞二磷胆碱组造模后14、21d与同一时间点对照组比较,差异无统计学意义(P>0.05)；康复训练组、胞二磷胆碱结合康复训练组造模后14、21d行为学评分明显优于同一时间点对照组,差异有统计学意义(P<0.05),且胞二磷胆碱结合康复训练组降低更明显(P<0.01)。
3.免疫组化：
造模后7d胞二磷胆碱组、康复训练组S-100、β-tubulin的表达水平与同一时间点对照组比较,增加不明显(P>0.05)；造模后7d胞二磷胆碱结合康复训练组S-100、β-tubulin的表达水平与同一时间点胞二磷胆碱组、康复训练组、对照组增加明显,差异有统计学意义(P<0.01)；造模后14、21d时胞二磷胆碱组、康复训练组、胞二磷胆碱结合康复训练组S-100、β-tubulin的表达水平较对照组增加(P<0.05),且胞二磷胆碱结合康复训练组较胞二磷胆碱组、康复训练组增加更显著(P<0.01)。
结论
1.脑缺血损伤引起大鼠明显的神经功能缺损体征,胞二磷胆碱结合合理有序的康复训练能明显改善脑缺血大鼠运动功能。
2.胞二磷胆碱以及康复训练均能促进脑缺血大鼠神经损伤的恢复,两者结合应用效果更加显著。
3.胞二磷胆碱结合康复训练能促进脑缺血后神经功能恢复,其可能与脑缺血周围皮质S-100、β-tubulin的表达水平的上调有关。
Objectives
To observe the effect of citicoline in combination with rehabilitative training on motor function and the expression of S-100,β-tubulin after focal cerebral ischemia in rats, It also provides academic foundation for medicine in combination with rehabilitative training.
Methods
One-hundred and twenty male adult Sprague-Dawley rats(3 month old, n=100) were subjected to left middle cerebral artery occlusion (MCAO) with the suture occlusion and ninety-six rats consistent with the standard were randomly divided into four group:control group(n=24), citicoline group(n=24), rehabilitative training group(n=24), citicoline in combination with rehabilitative training group(n=24). Three day after reperfusion, the rats in citicoline group were received citicoline on dose of 500mg/Kg daily, those in rehabilitative training group were received motor training programs including balancing, grasping, rotating and walking exercise, those in citicoline in combination with rehabilitative training group were received citicoline and motor training programs, those in control group were not recevived any treatment. The behavioral tests of every group were evaluated at the 7th,14th,21st day after MCAO. And Immunohistochemistry staining method was used to detect the expression of S-100 andβ-tubulin of peri-ischemia cortex at the 7th,14th,21st day after MCAO respectively.
With SPSS 13.0 statistical software, and the differences among the groups were compared with the One-Way ANOVA and Newman-Keuls test. A value of P<0.05 was considered statistically significant.
Results
1.The rat of every group appear the sign of nervous function defect after MCAO successful.
2. neuroethology assessment
As compared with the control group, citicoline group at the 7th,14th,21st day after MCAO and rehabilitative training group, citicoline in combination with rehabilitative training group at the 7th day after MCAO were no significant difference (P>0.05), but rehabilitative training group, citicoline in combination with rehabilitative training group at the 14th,21st day after MCAO(P<0.05) significantly superior to the control group, especially in the citicoline in combination with rehabilitative training group (P<0.01).
3. Immunohistochemistry Staining Method
As compared with the control group, the expression of S-100 andβ-tubulin of citicoline group, rehabilitative training group at the 7th day after MCAO were no significant difference respectively(P>0.05), but the citicoline in combination with rehabilitative training group was significant higher than other group at the 7th day after MCAO(P<0.05); As compared with the control group, the other group respectively was significant higher at the 14th,21st day after MCAO(P<0.05), especially in the citicoline in combination with rehabilitative training group (P<0.01).
Conclusions
1.Citicoline in combination with reasonable and obviously rehabilitative training can significant improve motor functional recovery in rats.
2.Citicoline and rehabilitative training can promote the recover of nervous injury after focal cerebral ischemia in rats, the effect of combined modality therapy was more obvious.
3.the functional recovery may be partially attributed to the up-regulation of S-100 andβ-tubulin in peri-ischemia cortex.
研究胞二磷胆碱结合康复训练对脑缺血大鼠运动功能恢复及对钙结合蛋白、β微管蛋白表达的影响,为临床药物结合康复训练治疗脑血管病提供理论依据。
研究方法
按随机数字表法,将120只成年雄性SD大鼠采用Longa线栓法制作大鼠左侧大脑中动脉闭塞(MCAO)模型,选出造模成功后符合标准的96只大鼠随机分为对照组、胞二磷胆碱组、康复训练组、胞二磷胆碱结合康复训练组4组,每组24只。造模成功后3d,胞二磷胆碱组开始给予胞二磷胆碱(每天500mg/kg体重),康复训练组开始进行滚笼、平衡木、转棒、网屏等训练,胞二磷胆碱结合康复训练组既给予胞二磷胆碱,又给予康复训练,对照组不做任何干预。各组分别在造模后7、14、21d时进行行为学评估综合评分,同时在相应时间点,经灌流固定后断头取脑；各组取脑组织行免疫组化法观察缺血周围皮质S-100.β-tubulin的表达情况。采用SPSS13.0统计学软件进行单因素方差分析以及Newman-Keuls检验,P<0.05认为差异有统计学意义。
结果
1.大脑中动脉梗塞成功造模后各组大鼠均不同程度出现神经功能缺损的体征。
2.神经行为学评估：
各组大鼠随时间推移行为学评分均逐渐减小,胞二磷胆碱组、康复训练组、胞二磷胆碱结合康复训练组造模后7d行为学评分分别与同一时间点对照组比较,差异无统计学意义(P>0.05)；胞二磷胆碱组造模后14、21d与同一时间点对照组比较,差异无统计学意义(P>0.05)；康复训练组、胞二磷胆碱结合康复训练组造模后14、21d行为学评分明显优于同一时间点对照组,差异有统计学意义(P<0.05),且胞二磷胆碱结合康复训练组降低更明显(P<0.01)。
3.免疫组化：
造模后7d胞二磷胆碱组、康复训练组S-100、β-tubulin的表达水平与同一时间点对照组比较,增加不明显(P>0.05)；造模后7d胞二磷胆碱结合康复训练组S-100、β-tubulin的表达水平与同一时间点胞二磷胆碱组、康复训练组、对照组增加明显,差异有统计学意义(P<0.01)；造模后14、21d时胞二磷胆碱组、康复训练组、胞二磷胆碱结合康复训练组S-100、β-tubulin的表达水平较对照组增加(P<0.05),且胞二磷胆碱结合康复训练组较胞二磷胆碱组、康复训练组增加更显著(P<0.01)。
结论
1.脑缺血损伤引起大鼠明显的神经功能缺损体征,胞二磷胆碱结合合理有序的康复训练能明显改善脑缺血大鼠运动功能。
2.胞二磷胆碱以及康复训练均能促进脑缺血大鼠神经损伤的恢复,两者结合应用效果更加显著。
3.胞二磷胆碱结合康复训练能促进脑缺血后神经功能恢复,其可能与脑缺血周围皮质S-100、β-tubulin的表达水平的上调有关。
Objectives
To observe the effect of citicoline in combination with rehabilitative training on motor function and the expression of S-100,β-tubulin after focal cerebral ischemia in rats, It also provides academic foundation for medicine in combination with rehabilitative training.
Methods
One-hundred and twenty male adult Sprague-Dawley rats(3 month old, n=100) were subjected to left middle cerebral artery occlusion (MCAO) with the suture occlusion and ninety-six rats consistent with the standard were randomly divided into four group:control group(n=24), citicoline group(n=24), rehabilitative training group(n=24), citicoline in combination with rehabilitative training group(n=24). Three day after reperfusion, the rats in citicoline group were received citicoline on dose of 500mg/Kg daily, those in rehabilitative training group were received motor training programs including balancing, grasping, rotating and walking exercise, those in citicoline in combination with rehabilitative training group were received citicoline and motor training programs, those in control group were not recevived any treatment. The behavioral tests of every group were evaluated at the 7th,14th,21st day after MCAO. And Immunohistochemistry staining method was used to detect the expression of S-100 andβ-tubulin of peri-ischemia cortex at the 7th,14th,21st day after MCAO respectively.
With SPSS 13.0 statistical software, and the differences among the groups were compared with the One-Way ANOVA and Newman-Keuls test. A value of P<0.05 was considered statistically significant.
Results
1.The rat of every group appear the sign of nervous function defect after MCAO successful.
2. neuroethology assessment
As compared with the control group, citicoline group at the 7th,14th,21st day after MCAO and rehabilitative training group, citicoline in combination with rehabilitative training group at the 7th day after MCAO were no significant difference (P>0.05), but rehabilitative training group, citicoline in combination with rehabilitative training group at the 14th,21st day after MCAO(P<0.05) significantly superior to the control group, especially in the citicoline in combination with rehabilitative training group (P<0.01).
3. Immunohistochemistry Staining Method
As compared with the control group, the expression of S-100 andβ-tubulin of citicoline group, rehabilitative training group at the 7th day after MCAO were no significant difference respectively(P>0.05), but the citicoline in combination with rehabilitative training group was significant higher than other group at the 7th day after MCAO(P<0.05); As compared with the control group, the other group respectively was significant higher at the 14th,21st day after MCAO(P<0.05), especially in the citicoline in combination with rehabilitative training group (P<0.01).
Conclusions
1.Citicoline in combination with reasonable and obviously rehabilitative training can significant improve motor functional recovery in rats.
2.Citicoline and rehabilitative training can promote the recover of nervous injury after focal cerebral ischemia in rats, the effect of combined modality therapy was more obvious.
3.the functional recovery may be partially attributed to the up-regulation of S-100 andβ-tubulin in peri-ischemia cortex.
引文
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