针刺对大鼠脑出血神经细胞凋亡变性的影响及机制探究  

Effect and Mechanism of Acupuncture on Nerve Cell Apoptosis and Degeneration in Rats with ICH

作　　者：李丹 万思琦[4] 殷岳杉 朱曦 李龙 赵永厚 LI Dan;WAN Siqi;YIN Yueshan;ZHU Xi;LI Long;ZHAO Yonghou(Heilongjiang Mental Hospital,Harbin 150036, China;Fudan University, Shanghai 200433, China;The Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China;Dongzhimen Hosptial of Beijing University of Chinese Medicine,Beijing 100700, China;Beijing University of Chinese Medicine, Beijing 100029, China)

机构地区：[1]黑龙江神志医院,黑龙江哈尔滨150036 [2]复旦大学,上海200433 [3]北京中医药大学第三附属医院,北京100029 [4]北京中医药大学东直门医院,北京100700 [5]北京中医药大学,北京100029

出　　处：《针灸临床杂志》2021年第11期56-62,共7页Journal of Clinical Acupuncture and Moxibustion

基　　金：中国博士后科学基金,编号:2020M670940;黑龙江省博士后面上资助项目,编号:LBH-Z19034。

摘　　要：目的:通过针刺干预观察大鼠脑出血后神经细胞形态结构、凋亡变性的动态变化,探讨针刺治疗大鼠脑出血后对miR-34a-5p的影响及作用机制。方法:自体血脑内注射建立大鼠脑出血模型,随机分为假手术组、模型组、模型+针刺治疗组、模型+针刺治疗+阴性对照组和模型+针刺治疗+miR-34a-5p激动剂干预组,每组12只。每组再随机分为1 d、7 d两个亚组。建模12 h后针刺治疗,于1 d、7 d时间点对处死大鼠收集脑组织,HE染色观察脑组织病理学变化,TUNEL和NeuN荧光双染,Fluoro-Jade B染色观察神经元细胞凋亡变性情况。结果:模型组大鼠在脑出血损伤后出现不同程度的炎性反应和神经细胞凋亡坏死。针刺后炎性反应减轻,神经细胞凋亡和变性坏死的数目明显减少(P<0.05),在给予针刺治疗7 d后,TUNEL和NeuN荧光双染实验和Fluoro-Jade B染色实验中阳性细胞数进一步减少。与模型+针刺治疗组比较,激动剂干预组的神经细胞凋亡和变性坏死数目明显增多(P<0.05)。结论:针刺可能通过下调miR-34a-5p减轻神经炎症,促进ICH大鼠神经功能恢复,发挥神经保护作用。Objective:To observe the dynamic changes of neuronal cell morphology,apoptosis and degeneration after intracerebral hemorrhage(ICH)intervened by acupuncture,and to explore the effect and mechanism of acupuncture on miR-34a-5p after ICH in rats.Methods:ICH model was established by the injection of autologous blood into the brain.The rats were randomly divided into the sham operation group,the model group,the acupuncture group,the acupuncture combined with agomir-NC group and the acupuncture combined with miR-34a-5p agomir group,with 12 rats in each group.Each group was randomly divided into two subgroups of 1 d and 7 d.After 12 hours of modeling,acupuncture was applied.The brain tissues were collected on 1 d and 7 d of acupuncture treatment,and the pathological changes of brain tissues were observed by HE staining.TUNEL and NeuN fluorescent double staining were used to observe neuronal apoptosis,Fluoro-Jade B staining was applied to analyze neuronal degeneration and apoptosis.Results:The rats in the model group had different degrees of inflammatory reaction and neuronal apoptosis and necrosis after ICH injury.After acupuncture,the inflammatory response was reduced,and the number of apoptosis,degeneration and necrosis of nerve cells was significantly decreased(P<0.05).After 7 days of acupuncture treatment,TUNEL and NeuN fluorescent double staining experiment and Fluoro-Jade B staining experiment showed positive cells were further reduced.Compared to that in the acupuncture group,the apoptosis,degeneration and necrosis of nerve cells were significantly increased in the agonist intervention group(P<0.05).Conclusion:Acupuncture may reduce nerve inflammation by down-regulating miR-34a-5p,promote the recovery of nerve function in ICH rats and exert a nerve protective effects.

关 键 词：针刺 脑出血 神经细胞凋亡 Fluoro-Jade B染色 

分 类 号：R246.6[医药卫生—针灸推拿学]