Aging-related changes in the transcriptional profile of cerebrum in senescence-accelerated mouse (SAMP10) is remarkably retarded by acupuncture

Brain aging plays a pivotal role in senescence process and is related to cognition impairments and memory deficit and increases the risk of neurological disorders, but ideal therapeutic intervention has not been found. Here we report a dramatic effect of acupuncture, a powerful non-drug therapy way, on preventing changes in gene expression profiles with aging in senescence-accelerated mouse (SAMP10). We show that 48 genes in 588 genes examined display 2-fold or greater changes in gene expression with aging. However, we find that acupuncture can completely or partially prevent expression alterations of almost all these genes induced by aging. Our findings highlight a role of acupuncture as a potential intervention in anti-aging process and clinical therapy in future.     

SAMP10鼠脑衰老相关基因HSP86、HSP84的表达及针刺影响的研究

目的：探讨针刺延缓衰老的作用机制。方法：采用快速老化小鼠SAMP10、SAMR1为模型，运用RT-PCR和地高辛标记的非放射性Northern blot技术，观察8月龄SAMR1对照组、SAMP10对照组、SAMP10针刺组及SAMP10非穴位刺激组全脑、皮层和海马HSP84、HSP86基因表达的变化。结果：SAMP10对照组小鼠热休克蛋白HSP84、HSP86在全脑、皮层、海马中的表达下调，针刺后表达上调并趋向于正常组。结论：SAMP10小鼠脑衰老与热休克蛋白HSP84、HSP86基因表达异常有关，针刺可以通过调节HSP84、HSP86基因表达增强细胞保护、抑制细胞凋亡、抵抗氧化应激，从而起到延缓衰老的作用。     

针刺对快速老化脑萎缩及痴呆模型小白鼠(SAMP10)行为学影响的实验研究

[目的]研究“醒脑开窍”针法和“调补气血”针法对SAMP10行为学异常改变的影响。[方法]采用自然快速老化脑萎缩及痴呆模型小鼠SAMP10,于实验前后各进行1次穿梭箱实验 ,并记录小鼠遭受电击次数、电击时间及主动反应时间。[结果]实验前 ,6月龄P10与R1相比 ,各行为学指标均有显著性差异 (P<0.01) ;P10各组之间无显著性差异 (P>0.05) ;实验后 ,醒脑组、补气血组各项指标与P10对照组比较 ,均有明显改善 (P<0.05,或P<0.01) ,与R1比较 ,均有显著性差异 (P<0 .01) ,而两种针法间比较 ,无显著差异(P>0.05)。[结论]P10较同月龄R1学习记忆能力显著下降 ,是理想的脑萎缩及痴呆动物模型。“醒脑开窍”针法和“调补气血”针法可部分改善SAMP10的学习记忆能力     

针刺对快速老化小鼠SAMP10皮质衰老相关基因表达谱的影响

目的利用基因芯片技术研究“益气调血,扶本培元”针法对快速老化小鼠SAMP10皮质基因表达的影响。方法选取雄性8月龄快速老化小鼠SAMP10（分成针刺腧穴组,对照组）和同源正常老化小鼠SAMR1,采用cDNA microarray技术,分析伴随快速老化SAMP10皮质基因表达谱的变化以及针刺对其的影响。结果芯片结果显示,伴快速老化。P10皮质共有9类17个基因的表达发生变化,15个基因表达下调,2个基因表达上调,施予“益气调血,扶本培元”针法后,所有与P10快速老化相关基因的表达均被部分或全部的逆转,显示针刺具有特异性。结论“益气调血,扶本培元”针法能够良性调节脑老化相关基因的表达,并可能通过该种作用延缓脑衰老。     

针刺对快速老化骨质疏松小鼠P6骨代谢因子OPG、BMP-2的影响

目的:探讨针刺治疗骨质疏松的机制。方法:以快速老化骨质疏松小鼠P6(SAMP6)及正常同源抗快速老化小鼠R1(SAMR1)为模型,采用Western Blot等方法,观察SAMP6对照组、SAMP6针刺组、SAMP6非穴位刺激组和SAMR1对照组股骨护骨素(OPG)、骨形态发生蛋白-2(BMP-2)蛋白表达的变化。结果:SAMP6对照组股骨OPG、BMP-2表达显著下调(P<0.05);针刺后两者表达上调并趋于正常(P<0.05)。结论:SAMP6小鼠发生骨质疏松与OPG、BMP-2等骨代谢因子表达异常有关,针刺可通过促进成骨细胞形成并分泌骨代谢因子来刺激骨形成,降低骨转换率,达到改善骨质疏松的作用。     

快速老化痴呆鼠海马,皮质p38表达的增龄变化及针刺影响的实验研究

[目的]探讨针刺的作用机制.[方法]采用western blotting技术,以快速老化痴呆鼠(SAMP10)为材料,同源正常老化小鼠(SAMR1)为对照,观察2、4、6、8、10、12月龄海马和皮质中p38表达的增龄变化,以及"益气调血,扶本培元"针法对8月龄SAMP10海马和皮质p38表达的影响.[结果]1)p38表达在正常老化R1海马和皮质均随增龄呈平稳上升趋势.8月龄后较2月龄存在明显差异.而P10则呈波动上升趋势.海马中4月龄即升高,6月龄达峰值后下降,10月龄至低谷但仍显著高于2月龄,此后至12月龄又上升;皮质中2月龄即升高,4月龄达峰值后下降,6月龄至低谷后再次升高并于8月龄达峰值,此后至12月龄再度下降,但仍显著高于2月龄水平.2)与8月龄R1相比,p38在P10海马和皮质表达普遍增高."益气调血,扶本培元"针法具有腧穴特异性,可降低该蛋白在P10两个脑区的表达,使之接近R1的水平.[结论]1)p38表达在R1与P10两系相同脑区之间随增龄存在差异.2)p38的增龄变化与P10快速老化痴呆特征的时间段接近.3)"益气调血,扶本培元"针法可有效降低8月龄P10海马和皮质p38表达量.     

针刺对快速老化小鼠SAMP10氧化应激相关基因表达的影响

【目的】探讨快速老化小鼠SAMP10脑衰老的机制，揭示“益气调血、扶本培元”针法改善SAMP10脑衰老症状的部分机制。【方法】应用cDNA阵列技术(cDNA Array)，观察快速老化小鼠SAMP10与正常老化小鼠SAMR1前脑588个基因表达的差异，同时观察“益气调血、扶本培元”针法对上述基因表达的影响。【结果】SAMP10与SAMR1相比，谷胱甘肽S转移酶、热休克蛋白86、成纤维细胞生长因子8、NF-kappa-B转录因子p65亚单位4个与氧化应激反应有关的基因表达上调，而针刺可使这些基因的表达下调。【结论】SAMP10的快速老化可能与其脑内的氧化应激水平升高有关，针刺可改善氧化应激水平进而达到延缓脑衰老的作用。     

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.

     

针刺对快速老化小鼠SAMP10转录调节因子NF-E2、YB-1、LRG47的影响

目的:探讨针刺延缓衰老的机制。方法:采用快速老化小鼠(SAMP10)、正常老化小鼠(SAMR1)为模型,运用地高辛标记的非放射性Northernblot技术,观察8月龄SAMR1对照组、SAMP10对照组、SAMP10针刺组及SAMP10非穴位刺激组全脑、皮层和海马NF-E2、YB-1、LRG47基因表达的变化。结果:SAMP10对照组小鼠NF-E2、YB-1、LRG47在全脑、皮层、海马中的表达下调,针刺后表达上调并趋向于正常组。结论:SAMP10小鼠脑衰老与NF-E2、YB-1、LRG47基因表达异常有关,针刺可以通过调节NF-E2、YB-1、LRG47基因表达增强红细胞系统功能和细胞增殖,提高细胞抗菌免疫,从而起到延缓衰老的作用。     

针刺对快速老化模型鼠脑中衰老相关基因表达的调整作用

目的：探讨针刺调整作用在分子水平的反映。方法：以快速老化模型鼠（SAM）脑中衰老相关基因的表达为研究对象。应用由磁珠分离法纯化mRNA,SMART-PCRcDNA合成和AFLP-银染法结合形成的mRNA-AFLP分析技术，进行mRNA指纹检测，寻找衰老相关性差异表达的基因片段。并观察针刺对其表达水平的影响。结果：随机应用20对引物作选择性AFLP扩增，共发现42条带型较为清晰的SAMP，R两系组间差异表达带，其中，33条（约占78.6%)在P系相对于R系低表达，9条（约占21.4%)在P系相对于R系高表达，针刺SAMP10的不同腧穴，可使异常低表达的基因表达水平不同程度地上调，使异常高表达的基因表达水平不同程度地下调，其变化均趋同于正常老化的SAMR1。结论：针刺对衰老机体紊乱的分子网络能发挥整体调节作用。促使其恢复协调平衡，从而延缓衰老进程。     

快速老化小鼠SAMP8模型粪便代谢物和肠道菌群改变的研究

目的研究快速老化小鼠SAMP8模型老化过程中粪便代谢物和肠道菌群的变化。方法采用1H-NMR代谢组学和宏基因组学技术,探索衰老相关的代谢标志物和肠道菌群,并对代谢物与肠道菌群进行Pearson关联分析。结果 SAMP8小鼠粪便中指认出31种内源性代谢物,与SAMR1小鼠相比,13种代谢物发生显著改变。差异代谢物主要富集在4条代谢途径:苯丙氨酸、酪氨酸和色氨酸的生物合成,缬氨酸、亮氨酸和异亮氨酸的生物合成,苯丙氨酸代谢,组氨酸代谢。肠道菌群分析发现10月龄快速老化小鼠SAMP8肠道菌群多样性明显改变,10种菌群的相对丰度显著改变。相关性分析结果表明,Christensenellaceae菌科与苯丙氨酸、组氨酸、缬氨酸、异亮氨酸和尿苷酸均呈正相关;Dehalobacterium菌属与酪氨酸,Planococcaceae动球菌科与缬氨酸均呈负相关。结论通过研究快速老化小鼠SAMP8模型粪便代谢物和菌群的变化规律,为衰老机制及抗衰老药物研究提供实验依据。     

针刺对大鼠脑组织神经生物学基因表达的研究

目的:采用基因芯片技术,分析探讨针刺预处理抗脑缺血再灌注损伤作用的分子机制.方法:取雄性Wistar大鼠6只,3只为正常对照组,另3只为针刺预处理组.针刺组先行"肾俞""百会"针刺预处理,出针后0.5 h断头取脑.基因芯片技术进行基因表达差异分析.结果:针刺预处理后0.5 h,大鼠脑组织表达变化大于2倍的基因共265个,20个基因表达差异具有显著性意义(P＜0.05).其中8个基因表达上调,12个基因表达下调;涉及神经元信号传递类(离子通道、递质、受体、第二信使)基因11个,涉及转录调控类基因5个,涉及代谢酶类基因2个,涉及热休克反应类基因1个,涉及细胞骨架类基因1个.结论:一些针刺诱导的基因表达产物在针刺预处理抗脑缺血再灌注损伤过程中发挥了重要作用;针刺预处理后神经细胞功能状态的改变可能是其随后发挥抗脑缺血再灌注损伤作用的重要基础.     

Differential localization of pain-related neural responses during acupuncture stimulation using Blood Oxygen Level Dependent (BOLD) fMRI in a canine model

The objective of this study was to differentiate the neuronal responses, which was related or unrelated, to pain associated with acupuncture stimulation, and to localize the brain regions with response to stimulation that is unrelated to pain by using Blood Oxygen Level Dependent (BOLD) functional MRI (fMRI). BOLD fMRI was performed in six normal healthy beagle dogs, during placebo and verum acupuncture stimulations, at the right side of BL60 (KunLun) acupoint before and after local anesthesia of the acupoint. The order of the four sessions was placebo; verum acupuncture stimulation; before local anesthesia; and followed by the same stimulation after local anesthesia. One-sample t-test analysis was performed to localize the activated or deactivated areas, during both pre-anesthesia and post-anesthesia. In order to compare the pre-anesthesia to post-anesthetic responses, and placebo to verum acupuncture stimulation, within-subject analysis was performed. The post-anesthetic verum acupuncture stimulation resulted in increased activations in the left somatic afferent area I and II, right visual and auditory association area, and the descending reticular activating system of the brainstem. In addition, differential areas during post-anesthesia compared to that of the pre-anesthesia were in the left olfactory peduncle and descending reticular activating system of the brainstem. These results indicate that the areas of specific neural pathway are considered to be unrelated to the pain response during acupuncture stimulation.     

Changes of metallothionein 1 and 3 mRNA levels with age in brain of senescence-accelerated mice and the effects of acupuncture

The effects of aging and acupuncture on brain MT1 and MT3 mRNA levels in senescence-accelerated mice (SAMP10) and accelerated senescence resistant mice (SAMR1) were analyzed by Northern blot analysis. Both MT1 and MT3 mRNA levels in SAMR1 were increased significantly from birth to month 4 and decreased gradually thereafter. In SAMP10, the MT3 mRNA level followed the same pattern as in SAMR1 before month 4, then decreased from month 4 to 6, but was over-expressed and exceeded the previous level at month 8. The MT1 mRNA expression in SAMP10 showed a zigzag pattern. Of two groups of SAMP10 mice treated with acupuncture, the xingnao group (PC6 and Du26 as acupoints) and the zibuganshen group (BL18 and BL23 as acupoints), both showed a higher MT1 mRNA level and a lower MT3 mRNA level than the age-matched control group. Meanwhile, in both of the acupuncture groups, the ratios of MT3 to MT1 were down-regulated to the normal range. Overall, these results suggested that over-expression of MT3 mRNA and the increase in MT3 to MT1 ratios in SAMP10 were correlated with aging, and could be an important physiological and pathological event in the aging process. Acupuncture altered the expression levels of MT1 and MT3 mRNA and differences between the effects of the two stimulated acupoints were seen. Therefore, maintenance of the balance between MTs mRNA expression and correct MTs concentrations is crucial for brain-endocrine-immune response and normal aging. Acupuncture could improve zinc ion bioavailability, by maintaining the balance between MT1 and MT3 mRNA expression levels and might explain one of the mechanisms by which acupuncture treatments defer aging and treat some age-related neurodegenerative diseases.     

针刺四缝穴对厌食症患儿血清瘦素水平的影响

目的:观察针刺四缝穴对厌食症患儿血清瘦素水平的影响,揭示血清瘦素水平变化与小儿厌食之间的关系。方法:42名患儿随机分为治疗A组(15例),针刺四缝穴治疗,每周1次,3次为一疗程;治疗B组(15例),针刺四缝穴治疗,2周1次,3次为一疗程;药物组(12例),口服乳酸菌素片,每次1~2片,每天3次,4周为一疗程。结果:治疗A组有效率为93.3%,治疗B组有效率为93.3%,均高于药物组75.0%,差异具有非常显著性意义(P<0.01);治疗后3组患儿血清瘦素水平与治疗前比较都有非常显著性变化(P<0.01);A、B两组血清瘦素水平的改善均优于药物组(P<0.05)。结论:针刺四缝穴能促进患儿血清瘦素分泌,改善患儿厌食症状。     

黄芩素干预快速老化小鼠SAMP8肝脏组织的代谢组学研究

目的基于~1H-NMR代谢组学技术,研究黄芩素对快速老化小鼠SAMP8肝脏的保护作用并探索其作用机制。方法采用SAMP8小鼠作为快速老化模型,考察黄芩素(ig)对SAMP8小鼠肝脏的保护作用。小鼠随机分3组,对照组为SAMR1小鼠、模型组为SAMP8小鼠、给药组为SAMP8小鼠+黄芩素。对照组和模型组给予等量生理盐水,给药组给予200 mg/kg黄芩素。分别取各组小鼠肝脏组织,采用苏木精-伊红(HE)染色观察其肝组织损伤程度,并对肝脏组织进行~1H-NMR检测,结合多元统计分析探讨黄芩素抗衰老小鼠肝损伤的作用。结果肝脏指数以及HE染色结果显示,黄芩素能明显改善SAMP8小鼠的肝损伤情况。代谢组学分析共找到8个潜在生物标志物,主要涉及3条代谢通路,分别是丙氨酸、天冬氨酸、谷氨酸代谢,甘氨酸、丝氨酸、苏氨酸代谢和肌醇磷酸代谢。结论黄芩素对SAMP8小鼠的肝损伤具有保护作用,其机制涉及多靶点、多途径。     

针刺风池、天柱对椎-基底动脉供血不足的影响

目的:评价针刺风池、天柱穴对椎-基底动脉供血不足(VBI)的临床疗效.方法:将116例VBI患者随机分为两组,治疗组58例采用针刺风池、天柱穴治疗,对照组58例采用口服尼莫地平治疗.观察治疗前后临床症状及经颅多普勒超声检测左椎动脉、右椎动脉、基底动脉平均血流速度的变化情况.结果:治疗组的总有效率为89.66%,对照组的总有效率为86.21%.针刺治疗对改善症状及改善椎-基底动脉平均血流速度有显著的疗效,并优于尼莫地平组(P＜0.05).结论:针刺风池、天柱穴治疗椎-基底动脉供血不足疗效显著.     

醒脑开窍针法联合艾灸治疗术后尿潴留50例(英文)

Urinary retention is the common complication after the operations of cervical cancer, hemorrhoids, fistulas, caesarean section, rectal cancer, meningioma, uterine fibroids, lumbar sacral disorder, etc. The catheterization is the conventional approach in clinic to deal with this problem. But, it easily causes a urinary tract infection or urethral injury, which may bring many living troubles and pains to the patients. The writer had adopted acupuncture for consciousness restoring resuscitation in combination with moxibustion to treat 50 cases of postoperative urinary retention. The summary is as follows.     

醒脑开窍针法联合艾灸治疗术后尿潴留50例

Urinary retention is the common complication after the operations of cervical cancer,hemorrhoids,fistulas,caesarean section,rectal cancer,meningioma,uterine fibroids,lumbar sacral disorder,etc.The catheterization is the conventional approach in clinic to deal with this problem.But,it easily causes a urinary tract infection or urethral injury,which may bring many living troubles and pains to the patients.The writer had adopted acupuncture for consciousness restoring resuscitation in combination with moxibustion to treat 50 cases of postoperative urinary retention.The summary is as follows.     

音乐电针对SAMP8小鼠行为学及大脑皮层NEP、IDE表达的影响

目的：探讨音乐电针治疗阿尔茨海默病（AD）的可能作用机制。方法：将SAMP8小鼠随机分为模型组、药物组和音乐电针组,SAMR1小鼠作为正常组。药物组给予盐酸多奈哌齐0.92mg/kg灌胃给药;音乐电针组电针＂百会＂、＂印堂＂穴20min后,点刺＂水沟＂穴。采用Morris水迷宫测定小鼠学习记忆能力,免疫组化法检测小鼠大脑皮层NEP及IDE的表达。结果：音乐电针改善模型小鼠学习记忆能力与药物比较,无显著性差异（P〉0.05）,并能显著提高大脑皮层NEP及IDE表达（P〈0.05）。结论：音乐电针具有改善AD学习记忆障碍的作用,其机制可能与促进Aβ降解酶表达有关。