补肾活血针刺法对SAMP8小鼠学习记忆能力及海马内源性神经干细胞增殖的影响

目的观察补肾活血针刺法对阿尔茨海默病(AD)模型SAMP8小鼠行为学及海马内源性神经干细胞增殖的影响。方法将18只7月龄雄性SAMP8小鼠作为AD模型,随机分为针刺组、非穴组和模型组,另取同月龄雄性SAMR1小鼠6只作为正常组。针刺组小鼠选取"肾俞""百会""血海""膈俞"进行针刺干预,非穴组小鼠取双侧肋下固定非穴点进行针刺,模型组和正常组小鼠予相同程度捉抓刺激,连续干预8周。采用水迷宫实验检测小鼠学习记忆能力,免疫组化检测海马Brd U阳性细胞数。结果与正常组比较,模型组小鼠逃避潜伏期明显延长(P0.05),停留在原平台象限时间明显缩短(P0.05),Brd U阳性细胞数明显减少(P0.05);与模型组、非穴组比较,针刺组小鼠逃避潜伏期明显缩短(P0.05),停留在原平台象限时间明显延长(P0.05),Brd U阳性细胞数明显增加(P0.05)。结论补肾活血针刺法可能通过诱导海马内源性神经干细胞的增殖,改善AD模型SAMP8小鼠的学习记忆能力。     

Effect of acupuncture on hippocampal mitochondrial proteome expression in SAMP8 mouse model with Alzheimer disease

Objective

To observe the effect of acupuncture on the expression of mitochondrial proteome in hippocampus of senescence-accelerated mouse prone 8 (SAMP8) mice models with Alzheimer disease (AD), and to explore the possible protective mechanism of acupuncture on mitochondria.

Methods

Sixty 6-month-old male SAMP8 mice were randomly divided into an acupuncture at acupoint group, an acupuncture at non-acupoint group and a model group, 20 mice in each group. The 20 male senescence-accelerated mouse/resistance 1 (SAMR1) mice of the same age were used as a normal control group. Shenshu (BL 23), Baihui (GV 20), Xuehai (SP 10) and Geshu (BL 17) were selected for acupuncture intervention in acupuncture at acupoint group. After an 8-week intervention, mitochondrial tissues were extracted from the hippocampus. Differentially expressed proteins were identified by subcellular organelle proteomics. Western blot was used to verify the expressions of some related proteins in hippocampal mitochondria.

Results

Compared with the model group, there were 13 differentially expressed protein spots in the acupuncture at acupoint group, of which, 9 were up-regulated, including neurofilament light polypeptide (NFL), actin (cytoplasmic 1, database ID: ACTB), tubulin beta-2A chain (TBB2A), tropomodulin-2 (TMOD2), pyruvate dehydrogenase E1 component subunit beta (PDHE1-β), NADH-ubiquinone oxidoreductase 75 kDa subunit (database ID: NDUS1), heat shock cognate 71 kDa protein (HSC71), pyruvate dehydrogenase E1 component subunit alpha (PDHE1-α) and ATP synthase beta subunit (ATP-β); 4 were down-regulated, including glial fibrillary acidic protein (GFAP), pyruvate dehydrogenase phosphatase 1 (PDP1), mitochondrial-processing peptidase subunit alpha (MMP-α) and adenosine kinase (ADK). According to the information provided in the protein database, most of the differentially expressed proteins involve the regulation of mitochondrial function and structure. The expression levels of NFL and TBB2A in the normal control group and the acupuncture at acupoint group were significantly higher than those in the acupuncture at non-acupoint group (P<0.05). ATP-β and NDUS1 expression levels were significantly higher in the acupuncture at acupoint group than those in the acupuncture at non-acupoint group (P<0.05); there was no significant difference between the acupuncture at non-acupoint group and the model group (P>0.05).

Conclusion

Acupuncture may achieve the potential therapeutic effect on AD by regulating the structure and functional proteins of hippocampal mitochondria.

     

针刺对阿尔茨海默病模型小鼠行为学及线粒体分裂蛋白1、融合蛋白1表达和超微结构的影响

目的观察针刺对阿尔茨海默病(AD)模型小鼠行为学,大脑海马神经元线粒体分裂蛋白1(Fis1)、线粒体融合蛋白1(OPA1)表达及线粒体超微结构的影响,探讨针刺治疗AD的作用机制。方法 40只雄性SAMP8小鼠随机分为针刺组和模型组,每组20只。另取20只同月龄雄性正常老化模型小鼠(SAMR1小鼠)作为正常组,针刺组选取"肾俞""百会""血海""膈俞"进行干预。8周后,Morris水迷宫实验检测小鼠行为学,之后提取海马组织,Western blot检测小鼠海马线粒体相关蛋白Fis1和OPA1的表达,电镜观察小鼠海马神经元线粒体超微结构。结果与模型组比较,针刺组逃避潜伏期明显缩短(P0.05),其停留在原平台象限时间延长和穿越原平台次数明显增加(P0.05);针刺组Fis1表达明显减弱,OPA1表达明显增强(P0.05,P0.01);针刺组线粒体超微结构明显改善,线粒体体密度及面密度明显增加(P0.05)。结论针刺可能通过提高模型小鼠学习记忆能力、下调Fis1表达、上调OPA1表达和改善线粒体超微结构,达到治疗AD的作用。