针灸对AD模型大鼠海马神经元线粒体保护机制研究

Protective mechanism of acupuncture-moxibustion on hippocampal neuron mitochondria in rats with Alzheimer's disease

目的:观察针灸对阿尔茨海默病(AD)模型大鼠海马神经元线粒体超微结构及沉默信息调节因子1(SIR1)的影响,探讨其在保护线粒体过程中的可能有效机制。方法:将Wistar大鼠80只随机分为正常组、假手术组、模型组和针灸治疗组,每组20只。运用大鼠海马注射β淀粉样蛋白1-42多肽(Aβ1-42)的模型复制方法对后两组复制AD模型,针灸治疗组采用针刺加灸"百会"和"肾俞"穴(益肾调督针灸法),每日1次,7次为一疗程,共治疗2个疗程;其余各组相应处理后均常规饲养,不做任何处理。运用透射电镜、免疫组化和免疫印迹技术分别检测AD模型大鼠海马神经元线粒体的超微结构及SIR1的水平。结果:针灸治疗组大鼠海马神经元线粒体超微结构能够有效地改善,与模型组比较具有明显的差异;模型组海马中SIR1的平均光密度值(0.21±0.10)及灰度值(136.82±47.42)水平明显低于正常组(0.47±0.09,281.44±57.98,均P<0.01),而针灸治疗组(0.32±0.11,199.52±58.12)可明显上调SIR1的水平,与模型组比较差异具有统计学意义(P<0.05)。结论:"益肾调督"针灸治疗AD可能通过改善线粒体超微结构,上调SIR1水平,从而达到改善线粒体损伤、保护线粒体功能的目的。

Objective To observe effects of acupuncture and moxlbustlon on ultrastructure ann suent mlormation regulator 1 （SIR1） in hippocampal neuron mitoehondria in rats with Alzheimer＇s all.sense （ND） in order to explore its possible effective mechanism during the process of protecting mitochondria. Methods Eighty Wistar rats were randomly divided into a normal group, a sham operation group, a model group and an acupuncture group, 20 rats in each one. The AD model was established in the model group and acupuncture group with injection of Amyloid 13 Protein Fragment1-14 （A131-42） into the hippocampus. Acupuncture and moxibustion was applied at ＂Baihui＂ （GV 20） and ＂Shenshu＂ （BL 23） in the acupuncture group, once a day, seven days as a treatment course and totally two course were required. The rest groups were all fed with normal diet, without any treatment. The transmission electron microscopy, immunohistochemistry and immunoblotting technique were respectively adapted to measure ultrastructure and level of STR1 in hippocampal neuron mitochondria in AD rats. Results Compared with the model group, the ultrastructure in hippocampal neuron mitochondria in the acupuncture group was effec- tively improved. The average optical density of SIR1 in hippocampus was 0.21-＋0.10 and the gray value was 136.82~47. 42 in the model group, which were obviously lower than O. 47_＋0.09 and 281.44＋57.98 in the normal group （both P〈0.01）. However, levels of SIR1 in the acupuncture group, they were 0.32±0.11 and 199.52± 58.12, which were significantly increased compared with those in the model group （both P〈0.05）. Conclusion The reinforcing-kidney and regulating-Governor Vessel method of acupuncture and moxibustion for AD could improve ultrastructure of mitochondria and increase levels of SIR1 to achieve the aim of recovering injury of mitochondria and protecting function of mitoehondria.