针刺足三里穴对癫痫大鼠海马神经细胞凋亡的影响研究
|
摘要
目的通过针刺大鼠不同机能状态下的足三里穴,观察大鼠海马组织内神经细胞的早、晚期凋亡情况,以探讨足三里对大鼠海马神经细胞凋亡的影响。方法 30只SD雄性大鼠,按照随机原则,分为正常空白组、正常足三里组、模型空白组、模型足三里组共4组。其中模型组大鼠采用氯化锂-匹罗卡品(LiCl—PILO)癫痫模型。造模成功后,各组大鼠分别予以针刺干预,连续针刺10天后取材。大鼠海马组织经实验常规处理、染色,制备成单细胞悬液,后经流式细胞仪上机检测。结果与正常空白组相比,正常足三里组大鼠海马组织神经细胞早期细胞凋亡在针刺后升高,差异具有统计学意义(P<0.05),而晚期细胞凋亡的表达在针刺前后则无明显变化(P> 0.05);模型空白组与正常空白组相比,癫痫造模后,大鼠海马神经细胞早、晚期细胞凋亡表达皆升高,且差异具有统计学意义(P<0.05);模型足三里组与模型空白组相比,针刺前后大鼠海马神经细胞的早、晚期细胞凋亡表达皆无统计学意义(P> 0.05);正常足三里组与模型足三里组相比,大鼠海马神经细胞的早、晚期细胞凋亡表达皆具有统计学意义(P<0.05)。结论癫痫发生过程存在一定程度的细胞凋亡,针刺足三里穴未能起到调控癫痫发生的作用,其只对正常状态下海马神经细胞的早期凋亡具有特异性的调控作用。
Objective The effects of Zusanli on the apoptosis of rat hippocampal neurons were observed by acupuncture at Zusanli Point in different functional states of rats. Methods Thirty SD male rats were divided into four groups according to the random principle, including normal blank group, normal Zusanli group, model blank group and model Zusanli group. The model group rats were treated with lithium chloride-pilocarpine(LiCl-PILO) epilepsy model. After successful modeling, rats in each group were given acupuncture intervention, and were taken after continuous acupuncture for 10 days. Rat hippocampus tissue was routinely treated and stained to prepare a single cell suspension, which was then detected by flow cytometry.Results Compared with the normal blank group, the early apoptosis of hippocampal neurons in normal Zusanli group increased after acupuncture, the difference was statistically significant(P<0.05),and the expression of late apoptosis did not change significantly before and after acupuncture(P> 0.05). Compared with the normal blank group, the expression of apoptosis in rat hippocampal neurons was increased after epileptic modeling, and the difference was statistically significant(P<0.05). Compared with the blank model group, there was no significant difference in the expression of early and late apoptosis of hippocampal neurons in the model Zusanli group before and after acupuncture(P> 0.05). Compared with the normal blank group and model Zusanli group, the expression of early and late apoptosis in rat hippocampal neurons was statistically significant(P<0.05).Conclusion There is a certain degree of apoptosis in the process of epilepsy. Acupuncture at Zusanli can not regulate the occurrence of epilepsy. It only has a specific regulation effect on the early apoptosis of hippocampal neurons in normal state.
Objective The effects of Zusanli on the apoptosis of rat hippocampal neurons were observed by acupuncture at Zusanli Point in different functional states of rats. Methods Thirty SD male rats were divided into four groups according to the random principle, including normal blank group, normal Zusanli group, model blank group and model Zusanli group. The model group rats were treated with lithium chloride-pilocarpine(LiCl-PILO) epilepsy model. After successful modeling, rats in each group were given acupuncture intervention, and were taken after continuous acupuncture for 10 days. Rat hippocampus tissue was routinely treated and stained to prepare a single cell suspension, which was then detected by flow cytometry.Results Compared with the normal blank group, the early apoptosis of hippocampal neurons in normal Zusanli group increased after acupuncture, the difference was statistically significant(P<0.05),and the expression of late apoptosis did not change significantly before and after acupuncture(P> 0.05). Compared with the normal blank group, the expression of apoptosis in rat hippocampal neurons was increased after epileptic modeling, and the difference was statistically significant(P<0.05). Compared with the blank model group, there was no significant difference in the expression of early and late apoptosis of hippocampal neurons in the model Zusanli group before and after acupuncture(P> 0.05). Compared with the normal blank group and model Zusanli group, the expression of early and late apoptosis in rat hippocampal neurons was statistically significant(P<0.05).Conclusion There is a certain degree of apoptosis in the process of epilepsy. Acupuncture at Zusanli can not regulate the occurrence of epilepsy. It only has a specific regulation effect on the early apoptosis of hippocampal neurons in normal state.
引文
[1] 张金波,王淑秋,王芳芳,等.癫痫发作后神经元凋亡的多重调控机制[J].现代生物医学进展,2011,11(23):4592.
[2] 柳成刚,姜元辉,石栏.针刺百会、腰奇穴对癫痫大鼠血清IL-1β、IL-6表达及海马神经元凋亡因子的影响[J].针灸临床杂志,2016,32(6):78.
[3] 金泽,曹晓婷,王春英,等.穴位埋线对癫痫大鼠海马神经元细胞凋亡及氨基酸的影响[J].上海针灸杂志,2016,35(2):218.
[4] 梁繁荣,赵凌.经穴特异性研究进展[J].天津中医药,2014,31(10):577.
[5] André V,Dubé C,Francois J,et al.Pathogenesis and Pharmacology of epilepsy in the lithium-pilocarpine model[J].Epilepsia,2007,48:41.
[6] Racine RJ.Modification of seizure activity by electrical stimulation:areas[J].Electroencephalogr Clin Neurophysiol,1975.38(1):1.
[7] 余曙光,郭义.实验针灸学[M].上海:上海科学技术出版社,2014:151.
[8] 杨海涛.气街理论指导针刺对癫痫患者生活质量、抑郁状态、内分泌功能的影响[D].广州中医药大学硕士学位论文,2008.
[9] 齐涛,李佩芳,王尚祉.针灸治疗癫痫研究概况[J].中医药临床杂志,2016,28(2):276.
[10] 姜硕.足三里穴配伍应用古代文献研究[D].山东中医药大学硕士学位论文,2010.
[11] 狄忠.足三里针灸适宜病症古代文献研究[D].山东中医药大学硕士学位论文,2009.
[12] 汤立新.有“艾”更健康(十)——如何灸足三里穴[J].长寿,2014,56(12):56.
[13] 陈小丽,岳增辉,刘丽,等.足三里穴的古今应用与研究[J].针灸临床杂志,2016,32(7):80.
[14] 姜迎萍,刘浩,徐平,从功能磁共振成像技术探讨针刺足三里的中枢作用机制[J].四川中医,2007,25(1):98.
[15] Colciaghi F,Finardi A,Nobili P,et al.Progressive Brain Damage,Synaptic Reorganization and NMDA Activa～tion in a Model of Epileptogenic Cortical Dysplasia[J].PLo S One,2014,9(2):e89898
[16] 董升平.中药补脑止痫散对癫痫大鼠海马c-fos、PKCa表达和谷氨酸、γ-氨基丁酸含量的影响[D].黑龙江中医药大学博士学位论文,2013.
[17] 徐先伟,程为平.补脑止痫散对戊四氮慢性点燃癫痫大鼠模型c-fos、c-jun蛋白表达的影响[J].中医药信息,2013,30(2):39.
[2] 柳成刚,姜元辉,石栏.针刺百会、腰奇穴对癫痫大鼠血清IL-1β、IL-6表达及海马神经元凋亡因子的影响[J].针灸临床杂志,2016,32(6):78.
[3] 金泽,曹晓婷,王春英,等.穴位埋线对癫痫大鼠海马神经元细胞凋亡及氨基酸的影响[J].上海针灸杂志,2016,35(2):218.
[4] 梁繁荣,赵凌.经穴特异性研究进展[J].天津中医药,2014,31(10):577.
[5] André V,Dubé C,Francois J,et al.Pathogenesis and Pharmacology of epilepsy in the lithium-pilocarpine model[J].Epilepsia,2007,48:41.
[6] Racine RJ.Modification of seizure activity by electrical stimulation:areas[J].Electroencephalogr Clin Neurophysiol,1975.38(1):1.
[7] 余曙光,郭义.实验针灸学[M].上海:上海科学技术出版社,2014:151.
[8] 杨海涛.气街理论指导针刺对癫痫患者生活质量、抑郁状态、内分泌功能的影响[D].广州中医药大学硕士学位论文,2008.
[9] 齐涛,李佩芳,王尚祉.针灸治疗癫痫研究概况[J].中医药临床杂志,2016,28(2):276.
[10] 姜硕.足三里穴配伍应用古代文献研究[D].山东中医药大学硕士学位论文,2010.
[11] 狄忠.足三里针灸适宜病症古代文献研究[D].山东中医药大学硕士学位论文,2009.
[12] 汤立新.有“艾”更健康(十)——如何灸足三里穴[J].长寿,2014,56(12):56.
[13] 陈小丽,岳增辉,刘丽,等.足三里穴的古今应用与研究[J].针灸临床杂志,2016,32(7):80.
[14] 姜迎萍,刘浩,徐平,从功能磁共振成像技术探讨针刺足三里的中枢作用机制[J].四川中医,2007,25(1):98.
[15] Colciaghi F,Finardi A,Nobili P,et al.Progressive Brain Damage,Synaptic Reorganization and NMDA Activa～tion in a Model of Epileptogenic Cortical Dysplasia[J].PLo S One,2014,9(2):e89898
[16] 董升平.中药补脑止痫散对癫痫大鼠海马c-fos、PKCa表达和谷氨酸、γ-氨基丁酸含量的影响[D].黑龙江中医药大学博士学位论文,2013.
[17] 徐先伟,程为平.补脑止痫散对戊四氮慢性点燃癫痫大鼠模型c-fos、c-jun蛋白表达的影响[J].中医药信息,2013,30(2):39.