穴位埋线抗癫痫作用及对海马氨基酸类神经递质影响的实验研究
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摘要
目的:通过观察穴位埋线对实验性癫痫大鼠痫性发作行为、脑电图、海马区氨基酸类神经递质的影响,探讨穴位埋线的抗癫痫作用及机制。
方法:50只SD雄性大鼠随机分为空白组、模型组、埋线组、西药组、针刺组,每组10只。分组进行预处理3天,以腹腔注射青霉素钠造成大鼠急性癫痫模型(空白组除外),观察:①造模后70分钟内痫性发作的潜伏期和痫性发作行为表现,②造模后70～85分钟内Fz与Pz区脑电图电压、频率的变化,③造模后90分钟处死大鼠,分离海马,测定海马中γ-氨基丁酸(GABA)、谷氨酸(Glu)、天门冬氨酸(ASP)、丙氨酸(Ala)、甘氨酸(Gly)、牛磺酸(Tau)的含量,计算GABA/Glu值。
结果:
①埋线组、西药组、针刺组的痫性发作潜伏期均比模型组延长(P＜0.05或P＜0.01),其中埋线组最长,但与西药组、针刺组比较差异均无显著性意义(P＞0.05)。
②埋线组大鼠出现的痫性行为表现点头/甩尾、须动/头面抽动、前肢/后肢抽动、全身抽动、窜动的次数均少于模型组(P＜0.05或P＜0.01),西药组全身抽动的次数明显少于模型组(P＜0.01),针刺组全身抽动、窜动的次数均少于模型组(P＜0.05或P＜0.01)。埋线组须动/头面抽动次数明显少于西药组、针刺组(P＜0.01),针刺组窜动的次数少于埋线组(P＜0.05)。
③埋线组痫性发作程度比模型组轻(P＜0.05),而西药组、针刺组与模型组比较差异均无显著性意义(P＞0.05)。
④模型组Fz与Pz区的脑电图电压均显著高于空白组(P＜0.01),而埋线组、西药组、针刺组的电压均低于模型组(P＜0.05或P＜0.01),但此3组之间比较差异均无显著性意义(P＞0.05)。
⑤模型组造模后脑电频率A、频率B均减慢,而埋线组、西药组、针刺组的频率A均比模型组增快(P＜0.05或P＜0.01),埋线组、针刺组的频率B均比模型组增快(P＜0.05或P＜0.01),对频率A的影响以Pz区明显。埋线组、西药组、针刺组的脑电频率A、频率B分别比较差异均无显著性意义(P＞0.05)。
⑥模型组海马兴奋性氨基酸Glu、ASP的含量均高于空白组(P＜0.05或P＜0.01),而埋线组、西药组、针刺组均明显低于模型组(P＜0.01),但埋线组Glu的含量也低于西药组、针刺组(P＜0.05或P＜0.01)。
⑦模型组海马区抑制性氨基酸GABA、Ala、Gly的含量均高于空白组(P＜0.01),埋线组均高于模型组、西药组(P＜0.01);埋线组GABA、Gly的含量均高于针刺组(P＜0.01);针刺组Ala的含量高于西药组(P＜0.01)。模型组Tau的含量高于空白组(P＜0.01),而埋线组、西药组、针刺组Tau的含量均低于模型组(P＜0.05或P＜0.01),而均与空白组相近(P＞0.05),但3组间比较差异均无显著性意义(P＞0.05)。
⑧模型组海马GABA/Glu值高于空白组(P＜0.01),埋线组、西药组分别高于模型组(P＜0.01),但埋线组GABA/Glu值分别高于西药组、针刺组(P＜0.05或P＜0.01)。
结论:
①穴位埋线可延长癫痫发作的潜伏期、减少癫痫发作行为、减轻癫痫发作程度,具有显著的抗癫痫作用,并优于丙戊酸钠、针刺法。
②穴位埋线可明显降低癫痫大鼠脑电电压、增快脑电频率,抑制痫性放电,改善癫痫脑电图,其作用与丙戊酸钠、针刺法相同。
③穴位埋线的抗痫机制与增加癫痫大鼠海马抑制性氨基酸GABA、Ala、Gly的含量,降低兴奋性氨基酸Glu、ASP的含量,维持Tau的稳定,提高GABA/Glu值有关。
Objective : To investigate anti-epilepsy effect and mechanism of catgut implantation at acupoint, by means of observing the effects of catgut implantation at acupoint on epileptic seizure behaviors, electroencephalogram, hippocampal amino acid neurotransmitter in experimental epilepsy rats.
Methods 50 SD male rats were randomized into groups A(normal), B (model), C(treated with catgut implantation at acupoint), D(treated with sodium valproate), E(treated with acupuncture), 10 in each group. Except group A, the rats in the 4 groups received intraperitoneal injection of sodium penicillin to induce acute epilepsy model after 3 days pretreatment. The latent period and the performances of epilepsy seizures within 70 minutes after modeling were observed, the voltage and frequency of EEG in Fz and Pz zones within 70～85 minutes after modeling were detected. After 90-minute modeling, the rats were killed, hippocampus was separated, the contents of gamrna-amino butyric acid (GABA), glutanic acid (Glu), aspartic acid (ASP), alanine (Ala), glycine (Gly) and taurine (Tau) in rats hippocampus were determined, and the ratio of GABA and Glu was calculated.
Results:
①The latent period of epilepsy seizures of group CD and E was prolongered than group B(P<0.05 or P<0.01), and that of group C was the longest but there was no statistical significance of differences, compared with groups D and E (P>0.05).
②The every times of epileptic attack behavior in group C, including nodding / tail-flick, beard shaking / face tic, anterior / posterior limb tic, systemic tic and abruptly jump, were less than those in group B respectively (P<0.05 or P<0.01). The times of systemic tic in group D was obviously less than that in group B (P<0.01), and the times of systemic tic and abruptly jump in group E were obviously less than in group B (P<0.05 or P<0.01). The times of beard shaking / face tic in group C was less than in group D and E respectively (P<0.05 or P<0.01), but the times of abruptly jump in group E was less than in group C (P<0.05).
③The degree of epileptic attack in group C was lighter than in group B, but there were no statistical significance in both group D and E compared to group B (P>0.05).
④The voltage of Fz and Pz zones in group B increased obviously(P<0.01 compared with group A), and that in group C, D and E decreased (P<0.05 or P<0.01 compared with group B), but there was no statistical significance of differences among the three groups(P>0.05).
⑤The frequency A and B of brain wave in group B slowed down(compared with group A), but the frequency A in group C, D and E increased(P<0.05 or P<0.01 compared with group B), the frequency B in group C and E increased (P<0.05 or P<0.01 compared with group B). The effect to frequency A was marked in Pz zone. But the frequencies of A and B in group C, D and E had no statistical significance of differences respectively (P>0.05).
⑥The contents of Glu、ASP in Hippocampus due to excitatory amino acid (EAA) in group B were higher than those in group A(P<0.05 or P<0.01), those in group C, D and E decreased obviously(P<0.05 or P<0.01 compared with group B), but the content of Glu in group C was lower than group D and E respectively (P<0.05 or P<0.01).
⑦The contents of GABA、Ala、Gly in Hippocampus due to inhibitory amino acid (IAA) in group B were higher than those in group A(P<0.01), but those of group C increased ( P<0.01 compared with group B and D), GABA and Gly of group C were higher than group E (P<0.01), and Ala of group E increased ( P<0.01 compared with group D). The content of Tau in group B was higher than group A(P<0.01), but that in group C, D and E decreased respectively (P<0.05 or P<0.01 compared with group B) and were near to that of group A (P>0.05) ,but there was no statistical significance of differences respectively among group C, D and E (P>0.05) .
⑧The ratio of hippocampal GABA and Glu in group B was higher than that in group A (P<0.01), those in group C and D were higher than group B respectively (P<0.01), and that in group C was higher than group D and E respectively (P<0.05 or P<0.01).
Conclusions:
①Catgut implantation at acupoint can prolong the latent period of epilepsy seizures, diminish the epileptic attack behavior, lessen the degree of epileptic seizure, thus it has remarkable anti-epilepsy effect, which is better than sodium valproate and acupuncture.
②Catgut implantation at acupoint can obviously decrease the brain wave voltage, increase the brain wave frequency, suppress the epilepsy electric discharge, and improve epilepsy EEG, all of which are the same, compared with sodium valproate and acupuncture.
③The anti-epilepsy mechanism of Catgut implantation at acupoint may be related to the contents increase of GABA, Ala and Gly due to IAA, to the decrease of Glu and ASP due to EAA, to the stability of Tau, and to the enhancement of the ratio of GABA and Glu in epilepsy rats hippocampus.
方法:50只SD雄性大鼠随机分为空白组、模型组、埋线组、西药组、针刺组,每组10只。分组进行预处理3天,以腹腔注射青霉素钠造成大鼠急性癫痫模型(空白组除外),观察:①造模后70分钟内痫性发作的潜伏期和痫性发作行为表现,②造模后70～85分钟内Fz与Pz区脑电图电压、频率的变化,③造模后90分钟处死大鼠,分离海马,测定海马中γ-氨基丁酸(GABA)、谷氨酸(Glu)、天门冬氨酸(ASP)、丙氨酸(Ala)、甘氨酸(Gly)、牛磺酸(Tau)的含量,计算GABA/Glu值。
结果:
①埋线组、西药组、针刺组的痫性发作潜伏期均比模型组延长(P＜0.05或P＜0.01),其中埋线组最长,但与西药组、针刺组比较差异均无显著性意义(P＞0.05)。
②埋线组大鼠出现的痫性行为表现点头/甩尾、须动/头面抽动、前肢/后肢抽动、全身抽动、窜动的次数均少于模型组(P＜0.05或P＜0.01),西药组全身抽动的次数明显少于模型组(P＜0.01),针刺组全身抽动、窜动的次数均少于模型组(P＜0.05或P＜0.01)。埋线组须动/头面抽动次数明显少于西药组、针刺组(P＜0.01),针刺组窜动的次数少于埋线组(P＜0.05)。
③埋线组痫性发作程度比模型组轻(P＜0.05),而西药组、针刺组与模型组比较差异均无显著性意义(P＞0.05)。
④模型组Fz与Pz区的脑电图电压均显著高于空白组(P＜0.01),而埋线组、西药组、针刺组的电压均低于模型组(P＜0.05或P＜0.01),但此3组之间比较差异均无显著性意义(P＞0.05)。
⑤模型组造模后脑电频率A、频率B均减慢,而埋线组、西药组、针刺组的频率A均比模型组增快(P＜0.05或P＜0.01),埋线组、针刺组的频率B均比模型组增快(P＜0.05或P＜0.01),对频率A的影响以Pz区明显。埋线组、西药组、针刺组的脑电频率A、频率B分别比较差异均无显著性意义(P＞0.05)。
⑥模型组海马兴奋性氨基酸Glu、ASP的含量均高于空白组(P＜0.05或P＜0.01),而埋线组、西药组、针刺组均明显低于模型组(P＜0.01),但埋线组Glu的含量也低于西药组、针刺组(P＜0.05或P＜0.01)。
⑦模型组海马区抑制性氨基酸GABA、Ala、Gly的含量均高于空白组(P＜0.01),埋线组均高于模型组、西药组(P＜0.01);埋线组GABA、Gly的含量均高于针刺组(P＜0.01);针刺组Ala的含量高于西药组(P＜0.01)。模型组Tau的含量高于空白组(P＜0.01),而埋线组、西药组、针刺组Tau的含量均低于模型组(P＜0.05或P＜0.01),而均与空白组相近(P＞0.05),但3组间比较差异均无显著性意义(P＞0.05)。
⑧模型组海马GABA/Glu值高于空白组(P＜0.01),埋线组、西药组分别高于模型组(P＜0.01),但埋线组GABA/Glu值分别高于西药组、针刺组(P＜0.05或P＜0.01)。
结论:
①穴位埋线可延长癫痫发作的潜伏期、减少癫痫发作行为、减轻癫痫发作程度,具有显著的抗癫痫作用,并优于丙戊酸钠、针刺法。
②穴位埋线可明显降低癫痫大鼠脑电电压、增快脑电频率,抑制痫性放电,改善癫痫脑电图,其作用与丙戊酸钠、针刺法相同。
③穴位埋线的抗痫机制与增加癫痫大鼠海马抑制性氨基酸GABA、Ala、Gly的含量,降低兴奋性氨基酸Glu、ASP的含量,维持Tau的稳定,提高GABA/Glu值有关。
Objective : To investigate anti-epilepsy effect and mechanism of catgut implantation at acupoint, by means of observing the effects of catgut implantation at acupoint on epileptic seizure behaviors, electroencephalogram, hippocampal amino acid neurotransmitter in experimental epilepsy rats.
Methods 50 SD male rats were randomized into groups A(normal), B (model), C(treated with catgut implantation at acupoint), D(treated with sodium valproate), E(treated with acupuncture), 10 in each group. Except group A, the rats in the 4 groups received intraperitoneal injection of sodium penicillin to induce acute epilepsy model after 3 days pretreatment. The latent period and the performances of epilepsy seizures within 70 minutes after modeling were observed, the voltage and frequency of EEG in Fz and Pz zones within 70～85 minutes after modeling were detected. After 90-minute modeling, the rats were killed, hippocampus was separated, the contents of gamrna-amino butyric acid (GABA), glutanic acid (Glu), aspartic acid (ASP), alanine (Ala), glycine (Gly) and taurine (Tau) in rats hippocampus were determined, and the ratio of GABA and Glu was calculated.
Results:
①The latent period of epilepsy seizures of group CD and E was prolongered than group B(P<0.05 or P<0.01), and that of group C was the longest but there was no statistical significance of differences, compared with groups D and E (P>0.05).
②The every times of epileptic attack behavior in group C, including nodding / tail-flick, beard shaking / face tic, anterior / posterior limb tic, systemic tic and abruptly jump, were less than those in group B respectively (P<0.05 or P<0.01). The times of systemic tic in group D was obviously less than that in group B (P<0.01), and the times of systemic tic and abruptly jump in group E were obviously less than in group B (P<0.05 or P<0.01). The times of beard shaking / face tic in group C was less than in group D and E respectively (P<0.05 or P<0.01), but the times of abruptly jump in group E was less than in group C (P<0.05).
③The degree of epileptic attack in group C was lighter than in group B, but there were no statistical significance in both group D and E compared to group B (P>0.05).
④The voltage of Fz and Pz zones in group B increased obviously(P<0.01 compared with group A), and that in group C, D and E decreased (P<0.05 or P<0.01 compared with group B), but there was no statistical significance of differences among the three groups(P>0.05).
⑤The frequency A and B of brain wave in group B slowed down(compared with group A), but the frequency A in group C, D and E increased(P<0.05 or P<0.01 compared with group B), the frequency B in group C and E increased (P<0.05 or P<0.01 compared with group B). The effect to frequency A was marked in Pz zone. But the frequencies of A and B in group C, D and E had no statistical significance of differences respectively (P>0.05).
⑥The contents of Glu、ASP in Hippocampus due to excitatory amino acid (EAA) in group B were higher than those in group A(P<0.05 or P<0.01), those in group C, D and E decreased obviously(P<0.05 or P<0.01 compared with group B), but the content of Glu in group C was lower than group D and E respectively (P<0.05 or P<0.01).
⑦The contents of GABA、Ala、Gly in Hippocampus due to inhibitory amino acid (IAA) in group B were higher than those in group A(P<0.01), but those of group C increased ( P<0.01 compared with group B and D), GABA and Gly of group C were higher than group E (P<0.01), and Ala of group E increased ( P<0.01 compared with group D). The content of Tau in group B was higher than group A(P<0.01), but that in group C, D and E decreased respectively (P<0.05 or P<0.01 compared with group B) and were near to that of group A (P>0.05) ,but there was no statistical significance of differences respectively among group C, D and E (P>0.05) .
⑧The ratio of hippocampal GABA and Glu in group B was higher than that in group A (P<0.01), those in group C and D were higher than group B respectively (P<0.01), and that in group C was higher than group D and E respectively (P<0.05 or P<0.01).
Conclusions:
①Catgut implantation at acupoint can prolong the latent period of epilepsy seizures, diminish the epileptic attack behavior, lessen the degree of epileptic seizure, thus it has remarkable anti-epilepsy effect, which is better than sodium valproate and acupuncture.
②Catgut implantation at acupoint can obviously decrease the brain wave voltage, increase the brain wave frequency, suppress the epilepsy electric discharge, and improve epilepsy EEG, all of which are the same, compared with sodium valproate and acupuncture.
③The anti-epilepsy mechanism of Catgut implantation at acupoint may be related to the contents increase of GABA, Ala and Gly due to IAA, to the decrease of Glu and ASP due to EAA, to the stability of Tau, and to the enhancement of the ratio of GABA and Glu in epilepsy rats hippocampus.
引文
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