摘要
背景:电针治疗阿尔茨海默病有较好的临床疗效,但其作用机制尚不清楚。目的:观察电针对阿尔茨海默病模型大鼠海马神经元突触数密度、面密度、突触连接带的平均面积及海马CA3区超微结构的影响。设计:完全随机分组设计,对照实验。单位:四川省人民医院中医科,成都中医药大学针灸推拿学院。材料:选用24月龄雄性SD大鼠50只,体质量(480±20)g;3月龄雄性SD大鼠6只,体质量(250±15)g。通道式水迷宫:由黑色玻璃制成(2.1m×1.7m×0.6m),水深40cm,水温22～25℃。共设4个盲端。WQ1002F型Hans电针仪。方法:实验于2002-09/2003-06在成都中医药大学三级动物实验中心完成。①先后通过3次通道式水迷宫实验将老年SD大鼠分组:第1次水迷宫实验(针对6只青年大鼠)在造模前8d进行,共4d。得出青年大鼠平均逃避潜伏期。第2次水迷宫实验(针对50只老年大鼠)在造模前4d进行,得出老年大鼠平均逃避潜伏期。其中36只潜伏期<青年大鼠潜伏期均值+1倍标准差值为正常老年大鼠,从中随机选出12只,随机分为2组:对照组和假手术组,每组6只。对其余24只大鼠采用穹窿-海马伞切断术造成阿尔茨海默病模型。第3次水迷宫实验(针对造模的24只大鼠)在造模后第2天进行,从中选出潜伏期大于青年大鼠潜伏期均值+2倍标准差值的大鼠,即为合格的阿尔茨海默病模型,再从中随机筛选出12只,随机分为2组:模型组和电针组,每组6只。②造模后第6天开始对电针组进行治疗,选穴百会、涌泉、太溪、血海,用30号3.33cm毫针分别在“百会”斜刺0.5寸,“涌泉”、“太溪”、“血海”直刺0.3寸,连接电针仪进行治疗,施以连续波,频率20Hz,电压2～4V,强度以大鼠能安静耐受为度(约为2mA),留针30min,1次/d,连续治疗20d。对照组、假手术组(只在与造模大鼠相同位置暴露大脑皮质,不切断穹窿-海马伞)和模型组只进行固定,未予任何治疗。③治疗结束后,采用透射电镜观察大鼠海马CA3区超微结构,采用体视学方法计数突触的数量和突触连接带与测试线的交点数,求出能够较好反映突触形态可塑性变化的体视学参数突触的数密度、面密度和突触连接带的平均面积。④组间两两比较,方差齐性用t检验,非齐性用t检验。主要观察指标:各组大鼠突触数密度、面密度、突触连接带的平均面积比较。结果:最终符合要求进入结果分析的老年大鼠24只,每组6只。①超微结构:对照组和假手术组的突触密度大于模型组,而突触平均面积较小;电针后突触的密度较模型组明显增大,而突触的平均面积较小。②大鼠海马CA3区突触数密度、面密度:模型组和电针组明显低于对照组和假手术组(P<0.05～0.01),以模型组最低;假手术组与对照组差异不明显(P>0.05);电针组明显高于模型组(P<0.01)。③大鼠海马CA3区突触连接带平均面积:模型组和电针组明显大于对照组和假手术组(P<0.05～0.01),以模型组最高;假手术组与对照组差异不明显(P>0.05);电针组明显小于模型组(P<0.01)。结论:电针能有效地使阿尔茨海默病大鼠海马神经元突触形态得到一定程度的修复,阻止海马神经元突触的退变。
BACKGROUND： Eleetroaeupuneture has a good clinical effect on Alzheimer disease, but its mechanism remains unclear.
OBJECTIVE： To observe the effect of electroacupuncture on number of synaptic numeric density （Nv）, surface density （Sv） and average size of synaptic conjunction and uhrastructure in hippocampal CA3 area of neurons of model rats with Alzheimer disease （AD）.
DESIGN： Completely randomized grouping and controlled study.
SETTING： Department of Traditional Chinese Medicine of Siehuan Provincial People＇s Hospital; College of Acupuncture ＆ Moxibustion and Massage of Chengdu University of Traditional Chinese Medicine.
MATERIALS： A total of 50 male SD rats of 24-month old weighing （480±20） g and 6 male rats of 3-month old weighing （250±15） g were selected in this study. Passage water maze （2.1×1.7×0.6） m^3 was made of black glasses with 40 em deep water and 4 eaecums. WQ1002F Hans electrically heated distilling apparatus was used.
METHODS： The experiment was carried out at the Grade m Animal Experimental Center of Chengdu University of Traditional Chinese Medicine between September 2002 and June 2003. ①Old SD rats were grouped on the basis of water-maze test results. Firstly, 6 young rats were submitted to water-maze test at 8 days before modeling for 4 continuous days to obtain average escaping latency. Secondly, 50 old rats were accepted in the water-maze test at 4 continuous days before modeling to obtain average escaping latency. A total of 36 rats whose lateneies were less than average values plus one standard deviation of young rats were regarded as normal old rats. Among them, 12 rats were randomly divided into control group and sham operation group with 6 in each. Another 24 rats were transeeted at fornix-fimbria AD modeling. Thirdly, 24 modeling rats were all adopted in the water-maze test at 2 days after modeling. Rats whose lateneies were more than average value plus two standard deviation of young rats were chosen as AD model. Twelve AD models were randomly divided into model group and eleetroaeupuneture group with 6 in each. ② On the 6^th day after modeling, rats in eleetroaeupuneture group were aeupunctured at Baihui （Du20） of 0.5 inch in slope, Yongquan （K1）, Taixi （K3） and Xuehai （Spl0） of 0.3 inch in depth with No. 30 milli-needie （3.33 cm）; Then, electrically heated distilling apparatus was used with successive waves, 20 Hz in frequency and 2-4 V in voltage. Tolerant stress of rats at quiet state was regarded as the standard value （2 mA）, the needle was maintained for 30 minutes, and the acupuncture was done once a day for totally 20 successive days. Rats in control group sham operation group （cerebral cortex was exposured at the same site as model rats, and fomix-fimbria was not cut off） and model group were only fixed but treated with-nothing.③ After treatment, ultrastrueture in hippocampal CA3 area of rats was observed with transmission electron microscope; synapse numbers and cross-point numbers between synaptie conjunction and test line were counted with stereologieal technique; stereologieal parameters, such as numeric density （Nv）, area surface （Sv） and average size of synaptie conjunction, which could i＇efleet plastic changes of synaptie form, were calculated at the same time. ④ Differences between every two groups were compared with t test at regular variance and t ＇test at irregular variance.
MAIN OUTCOME MEASURES： Comparisons of numeric density （Nv）, surface density （Sv） and average size of synaptie conjunction.
RESULTS： A total of 24 old rats were involved in the final analysis with 6 in each group. ① Uhrastrueture： Synaptie density of control and sham operation group was higher than that of model group, andaverage area of synapse was smaller; synaptic density of eleetroaeupuneture group increased compared with that of model group, and average area of synapse was smaller. ② Numeric density and surface density in hippoeampal CA3 area in model and electroaeupuneture group were lower than those of control group and sham operation group （P 〈 0.05）. There was no significant difference between sham operation group and control group （P 〉 0.05）, and those of eleetroaeupuncture group were higher than those of model group （P 〈 0.01）. ③ Average size of synaptie conjunction in hippoeampal CA3 of model and eleetroaeupuneture group Was higher than that of control and sham operation group （P 〈 0.05）. There was no significant difference between sham operation group and control group （P 〉 0.05）, and the value of electroacupuneture group was lower than that of model group （P 〈 0.01）.
CONCLUSION： Eleetrotherapy can repair synaptie form and inhibit synaptic degeneration of hippoeampal neurons in AD rats.
出处
《中国临床康复》
CSCD
北大核心
2006年第27期187-189,F0003,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(39870956)~~
