氦氖激光与电针穴位刺激对新生大鼠脑缺血缺氧后海马神经元存活的影响(英文)

Impacts of He-Ne laser and electric acupuncture on hippocampal neuron survival in cerebral hypoxic ischemia in neonatal rats

背景:缺血缺氧性脑病(hypoxic-ischemicbraindamage,HIBD)是引起脑性瘫痪的常见病,目前对其治疗尚未有特效的方法。目的:探讨氦氖激光穴位照射与电针穴位刺激两种疗法对新生大鼠脑缺血缺氧后海马神经元存活、发育及其脑源性神经营养因子(brain-derivedneurotrophicfactor,BDNF)、胆碱乙酰基转移酶(cholineacetyltransferase,ChAT)表达的作用,比较与分析两种疗法作用的差异,探讨治疗HIBD的新方法。设计:完全随机设计,实验对照研究。地点与材料:研究的地点为第二军医大学和郑州大学解剖教研室。材料为58只7d龄Wistar大鼠(购自河南省实验动物中心),体质量12～15g,雌雄不拘。干预:随机分为4组:对照组、缺血缺氧组、缺血缺氧后氦氖激光穴位照射组(简称激光组)和缺血缺氧后电针穴位刺激组(简称电针组)。缺血缺氧组～电针组动物参照于晓虹的半球性脑缺血缺氧实验模型法制作左半球缺血缺氧模型,激光组-电针组均选择“大椎”和“百会”穴位分别给予激光照射与电针刺激,22d后取脑作组织切片,HE、Nissl染色以及BDNF和ChAT免疫组织化学染色。主要观察指标:光镜下观察神经元及其尼氏体,检测灰度值与记数ChAT和BDNF阳性细胞以及统计学处理。结果:①对照组海马神经元结构清晰,胞浆内充满深蓝色尼氏体(灰度值122.90±12.

BACKGROUND:Hypoxic-ischemic brain damage(HIBD) that often results in cerebral palsy continues to be a common disease in perinatal medicine. At present,no eff ective therapeutic strategies has yet been developed to counteract this conditio n. OBJECTIVE:To study the effect of He-Ne laser acupoint irradiation and electri c acupuncture on hippocampal neuron survival and development as well as the expr essions of brain-derived neurotrophic factor(BDNF) and choline acetyl transfera se(ChAT) in HIBD in neonatal rats.To compare and analysis the different effectiv eness of two treatments in order to explore the new method against HIBD. DESIGN:A completely randomized controlled study. SETTING and PARTICIPANTS:The study was conducted at the Department of Anatomy of the Second Military Medical University of Chinese PLA and Zhengzhou Universit y.Fifty-eight seven-day-old rats were obtained from the experimental animal c enter of Henan Province with a body mass from 12 to 15 g in either gender. INTERVENTIONS: Rats were randomly allocated into four groups: control group(I) that neither ligated nor exposed to hypoxia,HIBD group(II) that exposed to hypox ic-ischemic condition,laser irradiation group(Ⅲ) and electric acupuncture grou p(EA,Ⅳ).Left brain Hypoxic-ischemic injury model were established in all anima ls of Ⅱ-Ⅳ groups on basis of Yu Xiaohong's animal model of unilateral hypoxi a-ischemia in neonatal rats. Animals in group Ⅲ and Ⅳ were treated either w ith He-Ne laser acupoint irradiation(Ⅲ) or EA(Ⅳ) in the next day following hy poxia synchronously by choosing dazhui and baihui acupoints for the treatment.Fi fty-eight rats were subjected to perfusion-fixation 22 days later,the left bra ins were paraffin wax embedded,sectioned and stained by HE and Nissl staining as well as ChAT and BDNF immunohistochemical staining. MAIN OUTCOME MEASURES:The neurons and Nissl bodies were observed,the grey scal e values were measured and the number of ChAT and BDNF positive neurons was coun ted under microscope.Data were statistically analyzed. RESULTS:① The structure of neurons in hippocampus and cortex was clear in gr oup I,and the neuronal cytoplasm was full of dark-blue-colored Nissl bodies(gr ey scale value was 122.90±12.10);there were neuron degeneration and necrosis in group Ⅱ,a great number of Nissl bodies in surviving neurons were lost(grey sca le value was 188.31±10.43);The neuron degeneration,necrosis and the loss of Nis sl bodies reduced in laser Ⅲ or EA Ⅳ and grey scale value decreased respecti vely(130.56±6.16 and 135.19±7.00);②The number of ChAT and BDNF positive neuro ns was 29.74±4.85 and 13.42±5.56 respectively in hippocampus in groupⅠ;ChAT p ositive neurons decreased 13.96±7.62 and BDNF positive neurons heightened 21.93 ±5.12 in group Ⅱ;ChAT and BDNF positive neurons all increased significantly to 26.42±6.02 and 33.02±7.58 in hippocampus respectively in group Ⅲ;ChAT and BD NF positive neurons also increased to 25.54±5.05 and 27.63±7.15 respectively in group Ⅳ;③There were significant differences in grey scale and the number of positive neurons between control and each treatment group(P< 0.05); The variati on in group III was more obvious than that of group Ⅳ. CONCLUSION:He-Ne laser acupoint irradiation and EA have neuroprotective effec ts against hypoxic ischemia,which can promote the survival, development and func tion of hippocampal neurons.The impact of He-Ne laser acupoint irradiating is m ore significant than EA.The mechanism of the difference between the effectivenes s of these two treatments still requires further validation,although it can be p resumed that He-Ne laser not only has biological function as EA but also can im prove some restoring enzymes of injured neurons.