背景:中国传统医学针灸治疗缺血性脑损伤可提高脑的抗损伤能力并加快损伤修复.目的:观察百会、大椎两穴同时给予电针刺激后大鼠脑源性神经生长因子及胆碱乙酰转移酶的表达水平,探讨电针对缺氧缺血性脑损伤的保护作用.设计:随机对照实验. 单位:郑州师范高等专科学校生命科学系.材料:实验于2000-06/2001-09在郑州大学基础医学院人体解剖教研室完成.实验选取出生后7 d的清洁级Wistar大鼠50只,随机分为假手术组10只、模型对照组20只、电针治疗组20只.自制常压缺氧舱,40 cm×50 cm×60 cm,有两个2 em×2 cm的小孔与外界相通,钠石灰吸收舱内水分和CO2.方法:①模型建立及电针刺激:假手术组不造模,模型对照组及电针治疗组建立缺氧缺血模型.造模后两组大鼠在室温中恢复1～4 h后,进行低氧处理,即置于恒温37℃的常压缺氧舱内,以1.5 L/min的速度通人8 mL/L O2,920 mL/L混合气体,2 h后将动物取出,返回母鼠身边继续哺乳喂养.电针治疗组从造模后第2天开始,用一寸毫针,沿皮斜刺百会穴(顶骨正中)及大椎穴(第7颈椎与第1胸椎间,背部正中),两穴同时给予电针刺激,施以连续波,频率16Hz,强度10V,留针10 min,1次/d,10 d为1个疗程,共两个疗程,两个疗程间隔2 d.模型对照组动物造模后未进行任何治疗.②细胞记数:模型对照组及电针治疗组大鼠于缺氧缺血后22 d给予麻醉处死,取左侧脑组织制作石蜡切片,光镜下对各组切片的免疫阳性细胞进行记数.评估脑神经细胞功能状态,选择海马区进行胆碱乙酰转移酶阳性细胞数计数,每张脑片随机选取5个视野,求出阳性细胞的算术平均值.评估神经组织损伤修复作用,选择皮质和海马区脑源性神经生长因子阳性细胞数计数,方法同上.主要观察指标:新生大鼠电刺激后脑组织胆碱乙酰转移酶和脑源性神经生长因子免疫阳性细胞的表达.结果:①脑海马区胆碱乙酰转移酶免疫阳性细胞的表达:与假手术组比较,模型对照组明显降低,而电针治疗组则无明显变化[(24.46±8.24),(13.96±7.62),(25.54±5.05)个/视野,P＜0.05,P＞0.05];电针治疗组高于模型对照组(P＜0.05).②脑皮质和海马区脑源性神经生长因子免疫阳性细胞的表达:与假手术组比较,模型对照组和电针治疗组均明显升高[(14.14±6.11),(24.49±8.31),(31.35±9.92)个/视野,P均＜0.05;(13.42±5.56),(21.93±5.12),(27.63±7.15)个/视野,P均＜0.05];电针治疗组高于模型对照组(P＜0.05).结论:电针刺激使缺氧缺血动物中枢胆碱能神经系统处于积极活动状态,使脑源性神经营养因子数量上升增进缺氧缺血动物的神经修复功能.
揹景:中國傳統醫學針灸治療缺血性腦損傷可提高腦的抗損傷能力併加快損傷脩複.目的:觀察百會、大椎兩穴同時給予電針刺激後大鼠腦源性神經生長因子及膽堿乙酰轉移酶的錶達水平,探討電針對缺氧缺血性腦損傷的保護作用.設計:隨機對照實驗. 單位:鄭州師範高等專科學校生命科學繫.材料:實驗于2000-06/2001-09在鄭州大學基礎醫學院人體解剖教研室完成.實驗選取齣生後7 d的清潔級Wistar大鼠50隻,隨機分為假手術組10隻、模型對照組20隻、電針治療組20隻.自製常壓缺氧艙,40 cm×50 cm×60 cm,有兩箇2 em×2 cm的小孔與外界相通,鈉石灰吸收艙內水分和CO2.方法:①模型建立及電針刺激:假手術組不造模,模型對照組及電針治療組建立缺氧缺血模型.造模後兩組大鼠在室溫中恢複1～4 h後,進行低氧處理,即置于恆溫37℃的常壓缺氧艙內,以1.5 L/min的速度通人8 mL/L O2,920 mL/L混閤氣體,2 h後將動物取齣,返迴母鼠身邊繼續哺乳餵養.電針治療組從造模後第2天開始,用一吋毫針,沿皮斜刺百會穴(頂骨正中)及大椎穴(第7頸椎與第1胸椎間,揹部正中),兩穴同時給予電針刺激,施以連續波,頻率16Hz,彊度10V,留針10 min,1次/d,10 d為1箇療程,共兩箇療程,兩箇療程間隔2 d.模型對照組動物造模後未進行任何治療.②細胞記數:模型對照組及電針治療組大鼠于缺氧缺血後22 d給予痳醉處死,取左側腦組織製作石蠟切片,光鏡下對各組切片的免疫暘性細胞進行記數.評估腦神經細胞功能狀態,選擇海馬區進行膽堿乙酰轉移酶暘性細胞數計數,每張腦片隨機選取5箇視野,求齣暘性細胞的算術平均值.評估神經組織損傷脩複作用,選擇皮質和海馬區腦源性神經生長因子暘性細胞數計數,方法同上.主要觀察指標:新生大鼠電刺激後腦組織膽堿乙酰轉移酶和腦源性神經生長因子免疫暘性細胞的錶達.結果:①腦海馬區膽堿乙酰轉移酶免疫暘性細胞的錶達:與假手術組比較,模型對照組明顯降低,而電針治療組則無明顯變化[(24.46±8.24),(13.96±7.62),(25.54±5.05)箇/視野,P＜0.05,P＞0.05];電針治療組高于模型對照組(P＜0.05).②腦皮質和海馬區腦源性神經生長因子免疫暘性細胞的錶達:與假手術組比較,模型對照組和電針治療組均明顯升高[(14.14±6.11),(24.49±8.31),(31.35±9.92)箇/視野,P均＜0.05;(13.42±5.56),(21.93±5.12),(27.63±7.15)箇/視野,P均＜0.05];電針治療組高于模型對照組(P＜0.05).結論:電針刺激使缺氧缺血動物中樞膽堿能神經繫統處于積極活動狀態,使腦源性神經營養因子數量上升增進缺氧缺血動物的神經脩複功能.
배경:중국전통의학침구치료결혈성뇌손상가제고뇌적항손상능력병가쾌손상수복.목적:관찰백회、대추량혈동시급여전침자격후대서뇌원성신경생장인자급담감을선전이매적표체수평,탐토전침대결양결혈성뇌손상적보호작용.설계:수궤대조실험. 단위:정주사범고등전과학교생명과학계.재료:실험우2000-06/2001-09재정주대학기출의학원인체해부교연실완성.실험선취출생후7 d적청길급Wistar대서50지,수궤분위가수술조10지、모형대조조20지、전침치료조20지.자제상압결양창,40 cm×50 cm×60 cm,유량개2 em×2 cm적소공여외계상통,납석회흡수창내수분화CO2.방법:①모형건립급전침자격:가수술조불조모,모형대조조급전침치료조건립결양결혈모형.조모후량조대서재실온중회복1～4 h후,진행저양처리,즉치우항온37℃적상압결양창내,이1.5 L/min적속도통인8 mL/L O2,920 mL/L혼합기체,2 h후장동물취출,반회모서신변계속포유위양.전침치료조종조모후제2천개시,용일촌호침,연피사자백회혈(정골정중)급대추혈(제7경추여제1흉추간,배부정중),량혈동시급여전침자격,시이련속파,빈솔16Hz,강도10V,류침10 min,1차/d,10 d위1개료정,공량개료정,량개료정간격2 d.모형대조조동물조모후미진행임하치료.②세포기수:모형대조조급전침치료조대서우결양결혈후22 d급여마취처사,취좌측뇌조직제작석사절편,광경하대각조절편적면역양성세포진행기수.평고뇌신경세포공능상태,선택해마구진행담감을선전이매양성세포수계수,매장뇌편수궤선취5개시야,구출양성세포적산술평균치.평고신경조직손상수복작용,선택피질화해마구뇌원성신경생장인자양성세포수계수,방법동상.주요관찰지표:신생대서전자격후뇌조직담감을선전이매화뇌원성신경생장인자면역양성세포적표체.결과:①뇌해마구담감을선전이매면역양성세포적표체:여가수술조비교,모형대조조명현강저,이전침치료조칙무명현변화[(24.46±8.24),(13.96±7.62),(25.54±5.05)개/시야,P＜0.05,P＞0.05];전침치료조고우모형대조조(P＜0.05).②뇌피질화해마구뇌원성신경생장인자면역양성세포적표체:여가수술조비교,모형대조조화전침치료조균명현승고[(14.14±6.11),(24.49±8.31),(31.35±9.92)개/시야,P균＜0.05;(13.42±5.56),(21.93±5.12),(27.63±7.15)개/시야,P균＜0.05];전침치료조고우모형대조조(P＜0.05).결론:전침자격사결양결혈동물중추담감능신경계통처우적겁활동상태,사뇌원성신경영양인자수량상승증진결양결혈동물적신경수복공능.
BACKGROUND: Acupuncture in Chinese traditional medicine improves capacity of brain on resisting injury and accelerates injury repair in treatment of ischemic brain injury.OBJECTIVE: To observe the expressions of cerebral nerve growth factor (NGF) and choline acetyltransferase after simultaneous stimulation with electroacupuncture on Baihui (GV 20) and Dazhui (GV 14) so as to probe into the protection of electroacupuncture on hypoxia-ischemia brain injury.DESIGN: Randomized controlled experiment.SETTING: Department of Life Science in Zhengzhou Normal High Training School.MATERIALS: The experiment was performed in Human Anatomy Department of Basic Medical College of Zhengzhou University, in which, 50 cleangrade neonatal Wistar rats of 7 days old were employed and randomized into sham-operation group (10 rats), model control (20 rats) and electroacupuncture group (20 rats). Hypoxia cabin was self-made with constant pressure, 40 cm ×50 cm×60 cm in size, with two small holes of 2 cm ×2 cm for each to connect with the external. Soda lime was used to absorb moisture and CO2 in the cabin.The model was not prepared in sham-operation group. In model control and electroacupuncture group, hypoxia-ischemia model was set up. After modeling, the rats in two groups were recovered for 1 to 4 hours at room temperature; afterwards, hypoxia management was performed. The rats were placed in hypoxia cabin with constant pressure at constant temperature of 37 ℃, inputting O2 8 mL/L and mixed gas 920 mL/L, 1.5 L/minute; 2hours later, the rats were returned back to female rats for lactation continuously. In electroacupuncture group, on the 2nd day after modeling, a filiform needle of one cun was used to insert Baihui (GV 20) (midpoint of parietal bone) subcutaneously and Dazhui (GV 14) (between C7 and T1, on the midline of back). Electric stimulation was done simultaneously on two points with continuous wave, 16 Hz in frequency, 10V in intensity, retaining for 10 minutes, once per day, 10 days made 1 course, totally two courses at interval of 2 days. In model control, no any treatment was given group, 22 days after hypoxia and ischemia, the rats were anesthetized and sacrificed and brain tissue on the left side was collected to prepare paraffin slices. Immune positive cell was counted on slices of each group under optic microscope. For evaluation on the function of brain nerve cell, hippocampus was selected to count the positive cells of choline acetyltransferase. On each brain slice, 5 visual fields were randomized to calculate the average of positive cells. For evaluation on injury repair of nerve tissue, cortex and hippocampus were selected to count positive cells of cerebral NGF. The method was same as the above.MAIN OUTCOME MEASURES: Immune positive cell expressions of choline acetyltransferase and cerebral NGF in brain tissue after electric stimulation in neonatal rats.ferase in brain hippocampus: Compared with sham-operation group, that in model control was lower remarkably, but, there was no obvious change in electroacupuncture group [(24.46±8.24), (13.96±7.62), (25.54±5.05) pcs/visual field, P ＜ 0.05, P ＞ 0.05]; that in electroacupuncture group was of cerebral NGF in cerebral cortex and hippocampus: Compared with sham-operation group, that in both model control and electroacupuncture was increased remarkably [(14.14±6.11), (24.49±8.31), (31.35±9.92) pcs/visual field, P ＜ 0.05; (13.42±5.56), (21.93±5.12), (27.63±7.15) pcs/visual field, P ＜ 0.05], of which, that in electroacupuncture group was higher than model control (P ＜ 0.05).CONCLUSION: Electroacupuncture stimulates central cholinergic nerve system into positive active state in hypoxia-ischemia animals, increases cerebral nerve growth factor in quantity and enhances nerve repair of hypoxia-ischemia animals.
