Effect of acupuncture on neural cell apoptosis of cerebral palsy rats

doi:10.3969/j.issn.2095-4344.2016.27.012

Abstract

Abstract:

BACKGROUND: Acupuncture has been shown to impact cerebral blood flow, relieve cerebral edema, improve microcirculation and reduce neural cell apoptosis in animals with cerebral palsy.
OBJECTIVE: To further verify the effect of acupuncture on neuronal apoptosis in cerebral palsy rats.
METHODS: Forty-five Sprague-Dawley rats were randomly divided into sham group, model group and the acupuncture group, with 15 rats in each group. In the model and acupuncture groups, rat models of cerebral palsy were established by ligating the left common carotid artery. In the sham surgery group, the left common carotid artery was not ligated. Rats in the acupuncture group received acupuncture from 2 days after model induction, once a day, for 20 consecutive days. Rats in the sham surgery and model groups were left intact. At 21 days after surgery, neurological functions, pathomorphological changes and neuronal apoptosis were observed.
RESULTS AND CONCLUSION: (1) Motor function score was significantly lower in the model group than in the sham surgery group (P < 0.05). Motor function score was significantly higher in the acupuncture group than in the model group at 21 days (P < 0.05). (2) Hematoxylin-eosin staining: In the model group, cells were scattered. Inflammatory cell infiltration and cystis degeneration were found. The number of nerve cells was reduced. Degeneration, necrosis, gliosis and karyopyknosis were detected. Cavitation was visible in some cells. Cell bodies became small. The structure was not distinct or disappeared. In the sham surgery group, the morphology and structure of nerve cells were normal. In the acupuncture group, the changes in morphology and structure of nerve cells were found between the sham surgery and model groups. (3) The number of apoptotic cells was significantly greater in the model group than in the sham surgery group (P < 0.05). The number of apoptotic nerve cells was significantly higher in the acupuncture group than in the sham surgery group (P < 0.05), but significantly lower than in the model group (P < 0.05). (4) Results confirmed that acupuncture therapy can improve neurological function, reduce nerve cell apoptosis, and play a protective effect on nerve cells of rats with cerebral palsy.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

Key words: Acupuncture, Cerebral Palsy, Apoptosis, Tissue Engineering

CLC Number:

R318

Jie Xiao-su, Hou Yu-jin. Effect of acupuncture on neural cell apoptosis of cerebral palsy rats[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(27): 4036-4042.

Figures/Tables 3

2.1 造模成功动物数量及过程造模动物流程表见图1。 2.2 模型更接近人类新生鼠缺血缺氧后出现的神经病理特征与人类的表现类似。此类模型制作的方法之一是将动物置于极度缺氧环境中一定时间制成脑瘫模型。单纯结扎单侧颈总动脉后同侧大脑半球的血流量并不减少，然而在联合缺氧条件下该侧半球血流量明显减少并造成缺血性病理损害。此类模型制作方法相对简单，实验条件较容易控制重复性好，研究结果发现该模型动物脑室周围白质损伤与人类早产儿脑白质病理损伤相似[3]。 2.3 各组大鼠运动功能评分情况比较由表1可以看出：模型组大鼠第7天、第14天和第21天的运动功能评分均明显低于假手术组(P < 0.05)，表明大鼠脑瘫模型制作成功。针刺组大鼠第7天、第14天和第21天的运动功能评分也明显低于假手术组(P < 0.05)。假手术组的运动功能评分随着时间的延长没有明显变化，模型组和针刺组随着时间的延长运动功能评分有所增加，其中模型组增加不明显，针刺组第21天时运动功能评分增加比较明显，表明针刺治疗可以改善大鼠的运动功能评分，针刺组在第7天时的运动功能评分和模型组没有差别 (P > 0.05)，针刺组在第14天时的运动功能评分高于模型组，但两者之间比较差异无显著性意义(P > 0.05)，针刺组在第21天时的运动功能评分明显高于模型组，两者比较差异有显著性意义(P < 0.05)。表明针刺治疗可以改善脑瘫大鼠的运动功能评分。 2.4 各组大鼠大脑组织苏木精-伊红染色结果结果显示模型组大鼠大脑组织细胞排列紊乱，局部有炎症细胞浸润，见囊性变，神经细胞数量减少，并有变性坏死，胶质细胞增生，细胞核固缩，部分神经细胞有空腔形成，胞体缩小，结构模糊或者消失；假手术组大鼠大脑组织细胞排列整齐，层次清楚，细胞形态和结构正常，细胞染色深，密度大；针刺组大鼠大脑组织细胞形态和结构比较清晰，细胞变性坏死减轻，偶见有细胞核固缩。见图2。 2.5 各组大鼠大脑组织神经细胞凋亡数目由表2可以看出在手术后第21天时，模型组的神经细胞凋亡数目明显高于假手术组(P < 0.05)；针刺组的神经细胞凋亡数目也明显高于假手术组(P < 0.05)；但针刺组的神经细胞凋亡数目明显低于模型组，两者比较差异有统计学意义(P < 0.05)。表明针刺治疗可以减少大鼠神经细胞的凋亡。"

References

[1] Rice, etal.The influence of immaturity on hypoxic- ischemic brain damage in the rat. Ann Neurol. 1981; 9(2):131-141.
[2] 李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:329.
[3] 李登宇,李洪涛,潘志华,等.脑瘫动物模型研究进展[J].中国康复,2010,25(3):227-229.
[4] Tan JL,Zheng XR,Zhang SS,et al.Response of the sensorimotor cortex of cerebral palsy rats receiving transplantation of vascular endothelial growth factor 165-transfected neural stem cells . Neural Regen Res. 2014;9 (19): 1763-1769.
[5] 付强,祁岩超,唐纯志,等. 针刺和神经干细胞联合治疗脑瘫幼鼠的效果评价[J]. 湖南中医药大学学报, 2011, 31(6): 51-53.
[6] 刘颖,邹丽萍. 电针对缺氧缺血性脑损伤幼鼠大脑皮质血流的影响[J]. 实用儿科临床杂志, 2011, 26(3): 198-200.
[7] 迟瑛娇,李晓捷,李莹. 针刺对宫内感染致脑损伤仔鼠脑组织胶质纤维酸性蛋白表达的影响[J]. 实用儿科临床杂志, 2007,22(15): 1168-1169.
[8] 孔妍,唐强,朱路文,等.缺氧缺血性脑损伤后针康法对幼鼠学习记忆能力及海马微管相关蛋白-2 表达的影响[J]. 中国康复理论与实践,2011, 17(4): 310-312.
[9] 闫丙川,刘合玉,贾天明,等.针刺对缺氧缺血性脑损伤大鼠保护作用的实验研究[J].神经疾病与精神卫生, 2005, 5(1): 33-35.
[10] Liu Y,Zou LP,Du JB,et al.Electro- acupuncture protects against hypoxic- ischemic brain-damaged immature rat via hydrogen sulfide as a possible mediator. Neurosci Lett.2010;485(1): 74-78.
[11] 丁春华,刘焕荣,张少丹,等. 针灸治疗幼鼠缺血缺氧性脑病的实验研究[J]. 中国应用生理学杂志, 2005, 21(4): 400-401.
[12] 吴至凤,温恩懿,赵聪敏,等. 针刺对发育期脑损伤大鼠脑组织细胞凋亡及Bcl-2、Bax蛋白表达的影响[J]. 重庆医学, 2010,39(21): 2898-2900.
[13] 黄雄昂,刘继明,唐绍灿.电针对窒息脑瘫幼鼠海马、纹状体和运动皮质GAP-43表达的影响[J].山东大学学报:医学版,2009, 47(9):44-48．
[14] 祁岩超,刘振寰,柴铁劬,等.针刺抑制幼鼠HIBD脑组织神经细胞凋亡的实验研究[J].中国实用神经疾病杂志, 2011, 14(15):1-4.
[15] Li MK,Li YJ,Zhang CF,et al.Acupuncture for ischemic stroke: cerebellar activation may be a central mechanism following Deqi. Neural Regen Res. 2015; 10 (12): 1997-2003
[16] Zou W,Chen QX,Sun XW,et al.Acupuncture inhibits Notch1 and Hes1 protein expression in the basal ganglia of rats with cerebral hemorrhage. Neural Regen Res. 2015; 10(3): 457-462.
[17] 马晓丽,张丽华,张士岭,等. EP对HIBD 新生大鼠脑组织BCL-2及BAX 表达的影响[J].中国伤残医学, 2012, 20(1): 24-25.
[18] 刘波,唐强,邢艳丽,等.头穴丛刺结合环境刺激对缺氧缺血性脑损伤新生大鼠海MAP-2和p38 表达影响的实验研究[J].针灸临床杂志, 2011, 27(2): 63-65.
[19] 刘振寰,潘佩光,赵勇,等. 电针对脑性瘫痪幼鼠脑组织单胺类神经递质的影响[J].中国中医基础医学杂志, 2007, 13(8):619-620.
[20] 杨进廉,宋军,杨金蓉.不同针刺部位对痉挛大鼠模型脑及脊髓内神经递质含量的影响[J]. 针灸临床杂志, 2008, 24(2): 37-39.
[21] 王风波,李晓捷,唐明薇. 电针对先天性脑损伤仔鼠脑组织脑源性神经生长因子表达的影响[J].中国中西医结合儿科学,2011,3(3): 213-215.
[22] Hengarter M O.The biochemistry of apoptosis. Nature. 2000;407:770-776.
[23] Cho SH,Chung KS,Choi JH,et al. Compound K, a metabolite of ginseng saponin,induces apoptosis via caspase-8-dependent pathway in HL-60 human leukemia cells.BMC Cancer.2009:449.
[24] 佟俊杰,张广耘,袁晓.细胞凋亡检测方法的研究进展[J].口腔医学,2010,30(7):437-439.
[25] HuertaS,GouletEJ,Huerta-YePezS,LivingstonEH.Sereening and detection of apoptosis.JSurg Res.2007;139(l):143-56.
[26] 李春丰,江清林,庄树文,等.白毒鹅膏菌毒素诱导小鼠肝细胞凋亡的DNA凝胶电泳分析[J].信息科技, 2010,18(11): 5,19.
[27] 王占奎,张连城,赵淑华,等.“调神通络”针法对脑缺血大鼠大脑皮层蛋白激酶表达的影响[J].针刺研究, 2012, 37(4): 312-317.
[28] 张晓明,瞿涛,左朝,等.电针对局灶性脑缺血再灌注大鼠迟发性细胞凋亡的影响[J].湖北中医药大学学报, 2012, 14(4):3-5.
[29] 彭兴甫,彭英,朱德军,等.针灸对脑梗死大鼠患侧海马 CA1 区 Bcl-2 的影响及其与学习记忆的关系[J].西部医学,2010,22(12):2204-2206.
[30] 罗文舒,吴永刚,杨卓欣,等.针刺督脉对局灶性脑缺血大鼠脑组织 TNF-amRNA表达的影响[J].中国中医急症,2012, 21(4):572-573.
[31] 张毅敏,唐纯志,程少冰,等.针刺对脑挫伤大鼠脑组织坏死面积及脑细胞凋亡的影响[J].中医杂志,2010,51(6): 528-530.
[32] 吴至凤,温恩懿,赵聪敏,等.针刺对发育期脑损伤大鼠脑组织细胞凋亡及Bcl-2、Bax 蛋白表达的影响[J].重庆医学, 2010,39(21):2898-2900.
[33] 刘振寰,潘佩光,祁岩超,等.通督醒神针刺法对脑性瘫痪幼鼠脑组织神经细胞凋亡及神经生长因子蛋白表达的影响[J].中医药临床杂志,2010,22(1):36-40.
[34] 杨俊,黄远桂,徐江涛.电针预处理抑制癫痫持续状态诱发的海马区神经元凋亡研究[J].辽宁中医药大学学报, 2011, 13(3):104-106.
[35] 刘忠锦,刘丹阳,张海燕,等.电针对阿尔茨海默病大鼠学习记忆及海马基质金属蛋白酶-2,9 的影响[J].中国当代医药,2013,20(13):15-16.
[36] 朱晶,邵水金,崔国红,等.电针血清对β淀粉样蛋白损伤原代培养大鼠海马神经元的保护作用[J].中国中医药信息杂志,2013,20(10):38-40.
[37] 陆征宇,赵虹,汪涛,等.针刺对帕金森病小鼠行为学及纹状体神经元凋亡的影响[J].针刺研究,2012,37(3):187-190.
[38] 王彦春,程宇核,马骏,等.电针对帕金森病大鼠黑质细胞形态及凋亡的影响[J].针刺研究,2010,35(6):415-421.
[39] 赵宇辉,孙忠人,黄亮,等.针灸对震颤麻痹大鼠行为学和细胞的凋亡的影响[J]. 世界中西医结合杂志,2010,5(3): 209-211.
[40] 蔡兴慧,宋小鸽,张荣军,等.针刺对海洛因复吸大鼠脑神经元超微结构变化的影响[J].中国药物依赖性杂志, 2012, 21(1):26-29.
[41] 王念,张荣军,宋小鸽,等. 针刺对海洛因复吸大鼠脑神经细胞凋亡的影响[J].安徽中医学院学报,2013,32(5): 44-48.
[42] 潘贵书、金寰.电针对海洛因成瘾大鼠海马细胞凋亡的影响[J].遵义医学院学报,2011,34(3):230-232.
[43] 金寰,潘贵书,许家洲.电针对海洛因成瘾大鼠 PAG 内细胞凋亡和 caspase-3表达的影响[J].神经解剖学杂志, 2010,26(6):647-651.