针刺对慢性脑缺血大鼠三叉神经脊束核神经元钙通道Cav1.3表达及凋亡的影响
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摘要
目的:探讨针刺对慢性脑缺血大鼠三叉神经脊束核神经元钙通道Cav1.3表达及凋亡的影响。方法:雄性Wistar大鼠36只,随机分成针刺组、缺血对照组、假手术组,通过阻断左侧颈总动脉和双侧椎动脉的方法制作慢性脑缺血模型,针刺组大鼠电针刺激大鼠"四白"穴15天。采用免疫组织化学方法检测大鼠三叉神经脊束核钙通道Cav1.3蛋白的表达情况,TUNEL法检测神经元的凋亡情况,电镜下观察神经元超微结构的变化。结果:针刺组大鼠三叉神经脊束核中钙通道Cav1.3蛋白表达明显高于缺血对照组,但仍低于假手术组(P<0.05);神经元凋亡数量为缺血对照组>针刺组>假手术组(P<0.05)。电镜下,针刺组大鼠三叉神经脊束核神经元内可见有衣小泡出现。结论:针刺大鼠"四白"穴能提高慢性脑缺血大鼠三叉神经脊束核神经元钙通道Cav1.3的表达,减少神经元凋亡。
Objective: To investigate the effects of acupuncture on calcium channel Cav1.3 expression and neuronal apoptosis of spinal nucleus of trigeminal nerve of chronic cerebral ischemia rats. Methods: 36 male Wistar rats were divided randomly into acupuncture group, ischemia group and sham operation group. The chronic cerebral ischemia rats' model was made by occluding the left common carotid artery and bilateral vertebral arteries; and then the rats in acupuncture group were acupunctured the "Sibai acupoints" for 15 days. Immunohistochemical stain was used to detect the calcium channel Cav1.3 expression in spinal nucleus of trigeminal nerve, TUNEL to detect apoptosis of neurons, electron microscope to observe the ultrastructure of neurons. Results: The calcium channel Cav1.3 protein expression in spinal nucleus of trigeminal nerve of rats in acupuncture group was obviously higher than ischemia group, but still lower than sham operation group(P<0.05). The apoptotic neurons number of three groups from high to low was ischemia group, acupuncture group and sham operation group(P<0.05). The coated vesicles could be found in neurons of rats in acupuncture group under electron microscope. Conclusions: Acupuncture of rats' "Sibai acupoints" can obviously up regulate the calcium channel Cav1.3 expression and decrease neuronal apoptosis in spinal nucleus of trigeminal nerve of chronic cerebral ischemia rats.
Objective: To investigate the effects of acupuncture on calcium channel Cav1.3 expression and neuronal apoptosis of spinal nucleus of trigeminal nerve of chronic cerebral ischemia rats. Methods: 36 male Wistar rats were divided randomly into acupuncture group, ischemia group and sham operation group. The chronic cerebral ischemia rats' model was made by occluding the left common carotid artery and bilateral vertebral arteries; and then the rats in acupuncture group were acupunctured the "Sibai acupoints" for 15 days. Immunohistochemical stain was used to detect the calcium channel Cav1.3 expression in spinal nucleus of trigeminal nerve, TUNEL to detect apoptosis of neurons, electron microscope to observe the ultrastructure of neurons. Results: The calcium channel Cav1.3 protein expression in spinal nucleus of trigeminal nerve of rats in acupuncture group was obviously higher than ischemia group, but still lower than sham operation group(P<0.05). The apoptotic neurons number of three groups from high to low was ischemia group, acupuncture group and sham operation group(P<0.05). The coated vesicles could be found in neurons of rats in acupuncture group under electron microscope. Conclusions: Acupuncture of rats' "Sibai acupoints" can obviously up regulate the calcium channel Cav1.3 expression and decrease neuronal apoptosis in spinal nucleus of trigeminal nerve of chronic cerebral ischemia rats.
引文
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[2]闫明茹,郑宇,孙洁,等.不同时机介入醒脑开窍针刺法对脑缺血再灌注大鼠SOD活性和MDA含量的影响[J].现代生物医学进展,2013,13(10):1887-1891.
[3]王春霞,孙远征.头穴丛刺法对慢性脑缺血大鼠海马血管内皮生长因子表达的影响[J].针刺研究,2012,37(5):375-379.
[4]孔祥玉,赵淑敏,张书琴.针刺过程中有衣小泡的形成、形态特征及其数量变化的研究[J].神经解剖学杂志,1993,9(1):39-45.
[5]Eckenhoxff MF,Pysh JJ.Double-walled coated vesicle formation:evidence for massive and transient conjugate internalization of plasma membranes during cerebellar development[J].J Neurocytol,1979,8(5):623-638.
[6]乔跃兵,马秀艳,孔祥玉,等.延长针刺时间大鼠三叉神经脊束核尾侧亚核非突触部位胞吐数量的变化[J].中国临床康复,2006,10(35):78-80
[7]Hirtz JJ,Boesen M,Braun N,et al.Cav1.3 calcium channels are required for normal development of the auditory brainstem[J].J Neurosci,2011,31(22):8280-8294.
[8]Nú?ez-Santana FL,Oh MM,Antion MD,et al.Surface L-type Ca2+channel expression levels are increased in aged hippocampus[J].Aging Cel,2014,13(1):111-120.
[9]王金华,刘佩芳.丹参酮ⅡA对脑缺血再灌注损伤大鼠神经元L-型钙通道表达的影响[J].中国中医药科技,2008,15(6):442-443.
[10]郭森,朱江,王永为,等.丁苯酞对缺血大鼠大脑皮质中钙通道Cav1.3表达的影响[J].中风与神经疾病杂志,2016,33(1):38-41.