针灸对脑性瘫痪模型幼鼠脑组织中氨基酸和神经递质的影响

Effect of acupuncture on cerebral amino acids and neurotransmitters in the immature rat model of cerebral palsy

背景:针灸对脑性瘫痪幼鼠有一定的治疗作用。目的:探讨火针对脑性瘫痪幼鼠脑组织中氨基酸和神经递质表达的影响。方法:取1周龄Wistar幼鼠50只,45只结扎左侧颈总动脉建立幼鼠脑性瘫痪模型,5只不处理作为对照组。45只模型幼鼠随机分为模型组、针刺组及阳性组,每组15只。于造模第3天开始,阳性组灌胃脑酶水解片溶液,火针组使用火针处理内关穴、涌泉穴、曲池穴、百会穴,治疗持续21 d。于末次治疗1 h后,进行悬挂实验及Y型迷宫实验,并处死大鼠分离其大脑皮质及海马区,组织匀浆后检测乙酰胆碱、去甲肾上腺素、5-羟色胺、谷氨酸和γ-氨基丁酸的表达变化,采用病理切片观察海马区组织结构变化。结果与结论:(1)一般情况:21 d内治疗组大鼠活动明显多于模型组。治疗21 d,与对照组相比,模型组体质量显著下降(P<0.05);与模型组相比,阳性组及火针组体质量均明显上升(P<0.05),且后2组差异无显著性意义;(2)悬挂实验及Y型迷宫实验:与对照组相比,模型组悬挂时间显著下降,犯错次数显著性上升(P<0.05);与模型组相比,阳性组及火针组悬挂时间显著性延长,阳性组和火针组的犯错次数显著下降(P<0.05);(3)神经递质水平:与对照组相比,模型组去甲肾上腺素、5-羟色胺和γ-氨基丁酸水平下降,谷氨酸水平增加(P<0.05);与模型组相比,阳性组和火针组的去甲肾上腺素、5-羟色胺和γ-氨基丁酸水平增加,谷氨酸水平减少(P<0.05);(4)苏木精-伊红染色:模型组可见大量的神经元结构破坏变形,体积减小,细胞核固缩,且与细胞膜边界不清晰;阳性组和火针组都可以在一定程度上治疗脑缺血引起的海马区脑细胞结构异常。(5)结果证明,火针法可以有效治疗幼鼠脑性瘫痪。

BACKGROUND： Acupuncture is proved to have therapeutic effect on cerebral palsy of immature rats. OBJECTIVE： To discuss the possible effect on cerebral amino acids and neuretransmitters in immature rats with cerebral palsy by using fire-needle. METHODS： Cerebral palsy models were established in 45 Wistar rats aged 1 week by left common carotid artery ligation and randomly assigned into model, acupuncture and positive groups （n=15 per group）. The other five rats were defined as controls, without any management. At 3 days, positive group rats were orally administrated with brain enzyme solution, and acupuncture group given fire-needle treatment at points of Neiguan （P 6）, Yongquan （K 1 ）, Quchi （LI 11） and Baihui （Du 20） for 21 days. Hanging test, inclined plane test and Y maze test were performed 1 hour after the last treatment, and the cerebral cortex and hippocampus were extracted and homogenized after execution. Lelves of acethl choline, norepinephrine, 5-hydroxytryptamine, glutamic acid and y-aminobutyric acid were recruited, and morphology of hippocampus was observed by pathological section. RESULTS AND CONCLUSION： （1） Within 21-day treatment, positive and acupuncture treatment rats were more active than model group. Compared with the control group, the body mass in the model group was significantly decreased （P 〈 0.05）; compared with the model group, the body mass in the positive and fire-needle groups were significantly increased （P 〈 0.05）, while it showed no significant difference between latter two groups. （2） The hanging time of model group rats was significantly shorter than that of control group （P 〈 0.05）, and significantly lengthened in the positive and fire-needle groups compared with the model group （P 〈 0.05）. Compared with the control rats, the error numbers of model group rats were significantly increased （P 〈 0.05）, and conversely, significantly decreased in the positive and fire-needle groups （P 〈 0.05）. （3） In the model group, the levels of norepinephrine, 5-hydroxytryptamine and y-aminobutyric acid ware significantly decreased, while glutamic acid significantly increased compared with the control group （P 〈 0.05）; compare with the model group, the positive and fire-needle group rats held significantly increased levels of norepinephrine, 5-hydroxytryptamine and y-aminobutyric acid and decrease of glutamic acid （P 〈 0.05）. （4） Hematoxylin-eosin staining showed that in the mode group, there were abundant neurons damaged, cell volume reduced, nucleus pycnosis and unclear border with nucleus both positive and fire-needle groups could improve neuron structure induced by cerebral ischemia. （5） In conclusion, fire-needle can treat cerebral palsy effectively.