脑皮质电刺激对氯化铁诱发大鼠癫痫的抑制作用
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摘要
癫痫是神经系统常见疾病,发病率约1%,全球共约5千万患者,我国约有900万患者。目前癫痫的治疗主要是药物和手术治疗,但仍然有约25%的患者无有效治疗,特别是癫痫灶定位于大脑功能皮质区的难治性癫痫患者是癫痫治疗的一大难点,因此寻求新的治疗方法十分必要。神经电刺激治疗癫痫是一种非常有前途的方法,刺激靶点包括迷走神经、小脑、尾状核、丘脑核团、黑质网状结构等,这些方法多为非特异性方法,直接进行癫痫灶刺激,特别是新皮质癫痫的脑皮质电刺激,是目前新的研究方向,但目前脑皮质电刺激治疗癫痫还仅仅是初步的探索,尚缺乏系统的研究。
本研究利用运动感觉区脑皮质注射氯化铁诱发急性和慢性癫痫大鼠模型,一方面通过检测发作频率和发作时程探讨脑皮质电刺激对氯化铁诱发大鼠急性癫痫发作的抑制作用,另一方面通过检测脑皮质后放电阈值、后放电时程和行为学评分探讨脑皮质电刺激对氯化铁诱发慢性癫痫大鼠模型脑皮质兴奋性的影响。实验结果如下:(1)脑皮质注射氯化铁诱发大鼠癫痫模型均出现程度不同的急性癫痫发作。(2)氯化铁诱发急性癫痫大鼠6h脑电图(electroencephalogram,EEG)记录中,平均发作次数1-Hz电刺激组(6.67±5.65)与对照组(18.33±6.93)相比有显著性差异(P<0.01),100-Hz电刺激组(8.33±5.38)与对照组相比有显著性差异(P<0.05)。(3)氯化铁诱发大鼠急性癫痫发作的平均发作时程1-Hz电刺激组(37.38±15.53s)和100-Hz电刺激组(39.60±11.82s)与对照组(61.43±16.61s)相比均有显著性差异(P<0.05)。(4)氯化铁诱发急性癫痫大鼠发作次数在1-Hz电刺激组和100-Hz电刺激组显示逐步下降的变化趋势,而对照组显示先短暂升高后随之下降的变化趋势。(5)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后测脑皮质后放电阈值低频低强度组(2.10±0.38mA)与对照组(1.50±0.33mA)相比组间有显著性差异(P<0.05),与脑皮质电刺激前初测脑皮质后放电阈值(1.55±0.35mA)相比组内有显著性差异(P<0.01)。低频高强度组、高频低强度组和高频高强度组脑皮质后放电阈值与对照组相比组间无统计学差异,但低频高强度组脑皮质后放电阈值终测值(1.85±0.35mA)与脑皮质电刺激前初测值(1.45±0.35mA)相比组内有显著性差异(P<0.05)。(6)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后行为学评分各组与对照组相比组间无统计学差异,但是低频低强度组行为学评分终测值(3.83±0.98)与脑皮质电刺激前初测值(4.83±1.17)相比组内有显著性差异(P<0.05)。(7)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后测后放电时程各组与对照组相比组间无统计学差异。(8)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后行为学评分与后放电阈值的比值低频低强度组(1.88±0.60)与对照组(3.22±0.67)相比组间有显著性差异(P<0.01),与脑皮质电刺激前初测值(3.22±1.05)相比组内有显著性差异(P<0.01)。低频高强度组(2.18±0.38)与对照组相比组间有显著性差异(P<0.05),与初测值(3.17±0.71)相比组内有显著性差异(P<0.01)。高频低强度组(2.50±0.33)和高频高强度组(3.66±0.73)与对照组相比组间无统计学差异,但高频高强度组终测值与初测值(3.10±0.37)相比组内有显著性差异(P<0.05)。
综合上述实验结果,本研究结论如下:(1)脑皮质注射氯化铁溶液后即给予1-Hz的方波脉冲脑皮质电刺激,可以明显降低氯化铁诱发大鼠急性癫痫发作的发作次数和发作时程。(2)脑皮质注射氯化铁溶液后即给予100-Hz的方波脉冲脑皮质电刺激,可以明显降低氯化铁诱发大鼠急性癫痫发作的发作次数和发作时程。(3)、重复的低频低强度或者低频高强度脑皮质电刺激可以升高氯化铁诱发慢性癫痫大鼠模型的脑皮质后放电阈值,提示脑皮质兴奋性降低,以低频低强度脑皮质电刺激的作用更明显。(4)、重复的高频高强度脑皮质电刺激后氯化铁诱发慢性癫痫大鼠模型的行为学评分与后放电阈值的比值较脑皮质电刺激前升高,提示脑皮质兴奋性升高。(5)、实验结果提示合适参数的脑皮质电刺激对氯化铁诱发大鼠癫痫具有抑制作用,为脑皮质电刺激治疗癫痫提供了新的依据。
Epilepsy is a common disorder of nervous system, which affects more than 50 million individuals worldwide and 9 million individuals in China—about 1% of the population. By now about 75% patients are controlled by antiepileptic drugs and operation. But there are at least 25% patients cannot be controlled by any available therapy, especially the patients whose epileptogenic cortex is overlapped with eloquent cortex. So the clinic treatment of epilepsy with epileptic focus overlapped with eloquent cortex is difficult and it is necessary to set up new therapies. Electric stimulation is an encouraging strategy for the treatment of intractable epilepsy. The targets of electric stimulation include vagus nerve, cerebellum, caudate nucleus, thalamic nuclei and reticular formation of substantia nigra. But the stimulation of targets above is non-specific. So it is maybe a good way to stimulate the epileptic focus directly. However, the effect of electric cortical stimulation on epileptic focus localized on the neocortex is not clear.
In this study, the rat models of acute and chronic focal epilepsy induced by intracortical injection of ferric chloride solution were used. In the first part, we studied the suppressive effects of the electric cortical stimulation on the seizures in the acute epileptic rat by investigated the frequency and duration of seizures induced by ferric chloride. In the second part, we studied the effects of the electric cortical stimulation on the cortical excitability in the chronic epileptic rat by investigated the threshold and duration of cortical afterdischarge and the behavior score. Results: (1) Spontaneous seizures were observed in all acute epileptic rats following intracortical injection of ferric chloride solution. (2) In 6h EEG recording, 1-Hz and 100-Hz electric cortical stimulation significantly decreased the number of seizures in the acute epileptic rat (6.67±5.65 and 8.33±5.38, respectively) compared with that of the control group (18.33±6.93, P<0.01 and P<0.05, respectively). (3) In 6h EEG recording, 1-Hz and 100-Hz electric cortical stimulation significantly decreased the duration of seizures in the acute epileptic rat (37.38±15.53s and 39.60±11.82s, respectively) compared with that of the control group (61.43±16.61s, P<0.05). (4) When we calculated the number of seizures in each hour in 6h EEG recording after electric cortical stimulation, we found that downtrend was observed in 1-Hz and 100-Hz electric cortical stimulation groups and transient upgrade followed downtrend was observed in control group. (5) After 1-Hz and 0.1mA repetitive electric cortical stimulation in 5d, the cortical afterdischarge threshold (2.10±0.38mA) significantly ascended in the chronic epileptic rat induced by intracortical injection of ferric chloride solution compared with that of the control group (1.5±0.33mA, P<0.05) and compared with the initial cortical afterdischarge threshold before electric cortical stimulation (1.55±0.35mA, P<0.01). There was no statistic difference of cortical afterdischarge thresholds after 1-Hz and 1.0mA, 100-Hz and 0.1mA, 100-Hz and 1.0mA repetitive electric cortical stimulation compared with that of the control group. But the cortical afterdischarge threshold after 1-Hz and 1.0mA repetitive electric cortical stimulation (1.85±0.35mA) significantly ascended compared with the initial cortical afterdischarge threshold before electric cortical stimulation (1.45±0.35mA, P<0.05). (6) There was no statistic difference of behavior score after repetitive electric cortical stimulation in 5d compared with that of the control group. But there was significantly difference of behavior score after 1-Hz and 0.1mA repetitive electric cortical stimulation (3.83±0.98) compared with the initial behavior score before electric cortical stimulation (4.83±1.17, P<0.05). (7) There was no statistic difference of duration of cortical afterdischarge after repetitive electric cortical stimulation in 5d compared with that of the control group. (8) The ratio of behavior score and cortical afterdischarge threshold after 1-Hz and 0.1mA (1.88±0.60) or 1-Hz and 1.0mA (2.18±0.38) repetitive electric cortical stimulation in 5d was significantly lower than that of the control group (3.22±0.67, P<0.01 and P<0.05, respectively). At the same time, the ratio of behavior score and cortical afterdischarge threshold after 1-Hz and 0.1mA or 1-Hz and 1.0mA repetitive electric cortical stimulation was significantly lower than initial ratio before electric cortical stimulation (3.22±1.05 or 3.17±0.71, respectively, P<0.01). There was no statistic difference of the ratio of behavior score and cortical afterdischarge threshold after 100-Hz and 0.1mA (2.50±0.33) or 1-Hz and 1.0mA (3.66±0.73) repetitive electric cortical stimulation in 5d compared with that of the control group. But the ratio of behavior score and cortical afterdischarge threshold after 100-Hz and 1.0mA repetitive electric cortical stimulation was significantly higher than initial ratio before electric cortical stimulation (3.10±0.37, P<0.05).
Conclusions: (1) 1-Hz square pulse electric cortical stimulation can significantly decreased the number and the duration of seizures in the acute epileptic rat just following the intracortical injection of ferric chloride solution. (2) 100-Hz square pulse electric cortical stimulation can also significantly decreased the number and the duration of seizures in the acute epileptic rat just following the intracortical injection of ferric chloride solution. (3) After the repetitive low frequency and low amplitude or low frequency and high amplitude electric cortical stimulation the cortical afterdischarge threshold significantly ascended in the chronic epileptic rat induced by intracortical injection of ferric chloride solution. It means the depression of cortical excitability after electric cortical stimulation, especially the repetitive low frequency and low amplitude electric cortical stimulation. (4) After the repetitive high frequency and high amplitude electric cortical stimulation the ratio of behavior score and cortical afterdischarge threshold was significantly higher than initial ratio before electric cortical stimulation. It means the rise of cortical excitability. (5) The results of this study showed that electric cortical stimulation with suitable parameter can produce a suppressive effect on the epileptic rat induced by intracortical injection of ferric chloride solution. The results of this study provide a new proof for the treatment of epilepsy by electric cortical stimulation.
本研究利用运动感觉区脑皮质注射氯化铁诱发急性和慢性癫痫大鼠模型,一方面通过检测发作频率和发作时程探讨脑皮质电刺激对氯化铁诱发大鼠急性癫痫发作的抑制作用,另一方面通过检测脑皮质后放电阈值、后放电时程和行为学评分探讨脑皮质电刺激对氯化铁诱发慢性癫痫大鼠模型脑皮质兴奋性的影响。实验结果如下:(1)脑皮质注射氯化铁诱发大鼠癫痫模型均出现程度不同的急性癫痫发作。(2)氯化铁诱发急性癫痫大鼠6h脑电图(electroencephalogram,EEG)记录中,平均发作次数1-Hz电刺激组(6.67±5.65)与对照组(18.33±6.93)相比有显著性差异(P<0.01),100-Hz电刺激组(8.33±5.38)与对照组相比有显著性差异(P<0.05)。(3)氯化铁诱发大鼠急性癫痫发作的平均发作时程1-Hz电刺激组(37.38±15.53s)和100-Hz电刺激组(39.60±11.82s)与对照组(61.43±16.61s)相比均有显著性差异(P<0.05)。(4)氯化铁诱发急性癫痫大鼠发作次数在1-Hz电刺激组和100-Hz电刺激组显示逐步下降的变化趋势,而对照组显示先短暂升高后随之下降的变化趋势。(5)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后测脑皮质后放电阈值低频低强度组(2.10±0.38mA)与对照组(1.50±0.33mA)相比组间有显著性差异(P<0.05),与脑皮质电刺激前初测脑皮质后放电阈值(1.55±0.35mA)相比组内有显著性差异(P<0.01)。低频高强度组、高频低强度组和高频高强度组脑皮质后放电阈值与对照组相比组间无统计学差异,但低频高强度组脑皮质后放电阈值终测值(1.85±0.35mA)与脑皮质电刺激前初测值(1.45±0.35mA)相比组内有显著性差异(P<0.05)。(6)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后行为学评分各组与对照组相比组间无统计学差异,但是低频低强度组行为学评分终测值(3.83±0.98)与脑皮质电刺激前初测值(4.83±1.17)相比组内有显著性差异(P<0.05)。(7)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后测后放电时程各组与对照组相比组间无统计学差异。(8)氯化铁诱发慢性癫痫大鼠模型重复脑皮质电刺激5d后行为学评分与后放电阈值的比值低频低强度组(1.88±0.60)与对照组(3.22±0.67)相比组间有显著性差异(P<0.01),与脑皮质电刺激前初测值(3.22±1.05)相比组内有显著性差异(P<0.01)。低频高强度组(2.18±0.38)与对照组相比组间有显著性差异(P<0.05),与初测值(3.17±0.71)相比组内有显著性差异(P<0.01)。高频低强度组(2.50±0.33)和高频高强度组(3.66±0.73)与对照组相比组间无统计学差异,但高频高强度组终测值与初测值(3.10±0.37)相比组内有显著性差异(P<0.05)。
综合上述实验结果,本研究结论如下:(1)脑皮质注射氯化铁溶液后即给予1-Hz的方波脉冲脑皮质电刺激,可以明显降低氯化铁诱发大鼠急性癫痫发作的发作次数和发作时程。(2)脑皮质注射氯化铁溶液后即给予100-Hz的方波脉冲脑皮质电刺激,可以明显降低氯化铁诱发大鼠急性癫痫发作的发作次数和发作时程。(3)、重复的低频低强度或者低频高强度脑皮质电刺激可以升高氯化铁诱发慢性癫痫大鼠模型的脑皮质后放电阈值,提示脑皮质兴奋性降低,以低频低强度脑皮质电刺激的作用更明显。(4)、重复的高频高强度脑皮质电刺激后氯化铁诱发慢性癫痫大鼠模型的行为学评分与后放电阈值的比值较脑皮质电刺激前升高,提示脑皮质兴奋性升高。(5)、实验结果提示合适参数的脑皮质电刺激对氯化铁诱发大鼠癫痫具有抑制作用,为脑皮质电刺激治疗癫痫提供了新的依据。
Epilepsy is a common disorder of nervous system, which affects more than 50 million individuals worldwide and 9 million individuals in China—about 1% of the population. By now about 75% patients are controlled by antiepileptic drugs and operation. But there are at least 25% patients cannot be controlled by any available therapy, especially the patients whose epileptogenic cortex is overlapped with eloquent cortex. So the clinic treatment of epilepsy with epileptic focus overlapped with eloquent cortex is difficult and it is necessary to set up new therapies. Electric stimulation is an encouraging strategy for the treatment of intractable epilepsy. The targets of electric stimulation include vagus nerve, cerebellum, caudate nucleus, thalamic nuclei and reticular formation of substantia nigra. But the stimulation of targets above is non-specific. So it is maybe a good way to stimulate the epileptic focus directly. However, the effect of electric cortical stimulation on epileptic focus localized on the neocortex is not clear.
In this study, the rat models of acute and chronic focal epilepsy induced by intracortical injection of ferric chloride solution were used. In the first part, we studied the suppressive effects of the electric cortical stimulation on the seizures in the acute epileptic rat by investigated the frequency and duration of seizures induced by ferric chloride. In the second part, we studied the effects of the electric cortical stimulation on the cortical excitability in the chronic epileptic rat by investigated the threshold and duration of cortical afterdischarge and the behavior score. Results: (1) Spontaneous seizures were observed in all acute epileptic rats following intracortical injection of ferric chloride solution. (2) In 6h EEG recording, 1-Hz and 100-Hz electric cortical stimulation significantly decreased the number of seizures in the acute epileptic rat (6.67±5.65 and 8.33±5.38, respectively) compared with that of the control group (18.33±6.93, P<0.01 and P<0.05, respectively). (3) In 6h EEG recording, 1-Hz and 100-Hz electric cortical stimulation significantly decreased the duration of seizures in the acute epileptic rat (37.38±15.53s and 39.60±11.82s, respectively) compared with that of the control group (61.43±16.61s, P<0.05). (4) When we calculated the number of seizures in each hour in 6h EEG recording after electric cortical stimulation, we found that downtrend was observed in 1-Hz and 100-Hz electric cortical stimulation groups and transient upgrade followed downtrend was observed in control group. (5) After 1-Hz and 0.1mA repetitive electric cortical stimulation in 5d, the cortical afterdischarge threshold (2.10±0.38mA) significantly ascended in the chronic epileptic rat induced by intracortical injection of ferric chloride solution compared with that of the control group (1.5±0.33mA, P<0.05) and compared with the initial cortical afterdischarge threshold before electric cortical stimulation (1.55±0.35mA, P<0.01). There was no statistic difference of cortical afterdischarge thresholds after 1-Hz and 1.0mA, 100-Hz and 0.1mA, 100-Hz and 1.0mA repetitive electric cortical stimulation compared with that of the control group. But the cortical afterdischarge threshold after 1-Hz and 1.0mA repetitive electric cortical stimulation (1.85±0.35mA) significantly ascended compared with the initial cortical afterdischarge threshold before electric cortical stimulation (1.45±0.35mA, P<0.05). (6) There was no statistic difference of behavior score after repetitive electric cortical stimulation in 5d compared with that of the control group. But there was significantly difference of behavior score after 1-Hz and 0.1mA repetitive electric cortical stimulation (3.83±0.98) compared with the initial behavior score before electric cortical stimulation (4.83±1.17, P<0.05). (7) There was no statistic difference of duration of cortical afterdischarge after repetitive electric cortical stimulation in 5d compared with that of the control group. (8) The ratio of behavior score and cortical afterdischarge threshold after 1-Hz and 0.1mA (1.88±0.60) or 1-Hz and 1.0mA (2.18±0.38) repetitive electric cortical stimulation in 5d was significantly lower than that of the control group (3.22±0.67, P<0.01 and P<0.05, respectively). At the same time, the ratio of behavior score and cortical afterdischarge threshold after 1-Hz and 0.1mA or 1-Hz and 1.0mA repetitive electric cortical stimulation was significantly lower than initial ratio before electric cortical stimulation (3.22±1.05 or 3.17±0.71, respectively, P<0.01). There was no statistic difference of the ratio of behavior score and cortical afterdischarge threshold after 100-Hz and 0.1mA (2.50±0.33) or 1-Hz and 1.0mA (3.66±0.73) repetitive electric cortical stimulation in 5d compared with that of the control group. But the ratio of behavior score and cortical afterdischarge threshold after 100-Hz and 1.0mA repetitive electric cortical stimulation was significantly higher than initial ratio before electric cortical stimulation (3.10±0.37, P<0.05).
Conclusions: (1) 1-Hz square pulse electric cortical stimulation can significantly decreased the number and the duration of seizures in the acute epileptic rat just following the intracortical injection of ferric chloride solution. (2) 100-Hz square pulse electric cortical stimulation can also significantly decreased the number and the duration of seizures in the acute epileptic rat just following the intracortical injection of ferric chloride solution. (3) After the repetitive low frequency and low amplitude or low frequency and high amplitude electric cortical stimulation the cortical afterdischarge threshold significantly ascended in the chronic epileptic rat induced by intracortical injection of ferric chloride solution. It means the depression of cortical excitability after electric cortical stimulation, especially the repetitive low frequency and low amplitude electric cortical stimulation. (4) After the repetitive high frequency and high amplitude electric cortical stimulation the ratio of behavior score and cortical afterdischarge threshold was significantly higher than initial ratio before electric cortical stimulation. It means the rise of cortical excitability. (5) The results of this study showed that electric cortical stimulation with suitable parameter can produce a suppressive effect on the epileptic rat induced by intracortical injection of ferric chloride solution. The results of this study provide a new proof for the treatment of epilepsy by electric cortical stimulation.
引文
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