中南大学湘雅三医院,湖南省长沙市河西岳麓区桐梓坡路138号, 410013
liruomeng1983@163.com
收稿:2025-01-18,
修回:2025-04-28,
纸质出版:2025-07-02
移动端阅览
任丹,张婷,徐江喜等.补肾活血针刺法对SAMP8小鼠海马组织线粒体膜电位及SIRT1/PGC-1α轴的影响[J].中医杂志,2025,66(13):1378-1385.
REN Dan,ZHANG Ting,XU Jiangxi,et al.Effects of Kidney-Tonifying and Blood-Activating Acupuncture on Mitochondrial Membrane Potential and SIRT1/PGC-1α Axis in Hippocampal Tissue of SAMP8 Mice[J].Journal of Traditional Chinese Medicine,2025,66(13):1378-1385.
任丹,张婷,徐江喜等.补肾活血针刺法对SAMP8小鼠海马组织线粒体膜电位及SIRT1/PGC-1α轴的影响[J].中医杂志,2025,66(13):1378-1385. DOI: 10.13288/j.11-2166/r.2025.13.013.
REN Dan,ZHANG Ting,XU Jiangxi,et al.Effects of Kidney-Tonifying and Blood-Activating Acupuncture on Mitochondrial Membrane Potential and SIRT1/PGC-1α Axis in Hippocampal Tissue of SAMP8 Mice[J].Journal of Traditional Chinese Medicine,2025,66(13):1378-1385. DOI: 10.13288/j.11-2166/r.2025.13.013.
目的
2
探讨补肾活血针刺法治疗阿尔茨海默病的可能作用机制。
方法
2
雄性SAMP8小鼠40只随机分为模型组、针刺组、非穴位组、多奈哌齐组,每组10只,SAMR1小鼠10只作为正常组。针刺组针刺小鼠“百会”“血海”“肾俞”和“膈俞”,其中“血海”“肾俞”和“膈俞”先刺激左侧穴位,次日再刺激右侧,交替进行,每日1次。非穴位组取双侧肋下固定非经非穴点进行针刺,每日1次。模型组及正常组小鼠进行与以上两组相同时间、程度的捉抓刺激。多奈哌齐组小鼠接受1 mg/kg多奈哌齐胃管灌胃治疗,每日1次。各组小鼠均干预4周。干预结束后,各组小鼠进行Morris水迷宫实验(记录定位航行潜伏期、穿越平台象限次数)及旷场实验(记录运动距离);HE及Nissl染色观察小鼠海马组织神经元细胞损伤情况;检测海马组织线粒体膜电位和活性氧(ROS)相对倍数;Western Blot法及qRT-PCR法分别检测沉默调节蛋白1(SIRT1)、过氧化物酶体增殖活化受体γ辅助活化因子1α(PGC-1α)蛋白水平及其mRNA表达。
结果
2
与正常组比较,模型组和非穴位组小鼠定位航行潜伏期及海马组织ROS相对倍数升高,穿越平台象限次数、旷场实验运动距离、海马组织Nissl染色阳性细胞数量、线粒体膜电位及SIRT1、PGC-1α蛋白水平和mRNA表达均降低(
P
<0.01);HE染色示小鼠海马组织神经元排列松散,数量减少,且空泡变性;Nissl染色示海马区锥体神经元不整齐,细胞质内Nissl体数量较少,染色较浅。与模型组比较,针刺组和多奈哌齐组小鼠定位航行潜伏期及海马组织ROS相对倍数降低,穿越平台象限次数、旷场实验运动距离、海马组织Nissl染色阳性细胞数量、线粒体膜电位及SIRT1、PGC-1α蛋白水平和mRNA表达均升高(
P
<0.01),且HE染色和Nissl染色显示海马组织病理损伤显著改善。与多奈哌齐组比较,针刺组小鼠定位航行潜伏期缩短(
P
<0.01)。
结论
2
补肾活血针刺法可激活海马组织SIRT1/PGC-1α信号通路,改善线粒体功能,从而缓解神经元损伤,是其治疗阿尔茨海默病的可能机制之一。
Objective
2
To explore the potential mechanisms of kidney-tonifying and blood-activating acupuncture
for Alzheimer's disease.
Methods
2
Male SAMP8 mice were randomly divided into a model group, acupuncture group, non-acupoint group, and donepezi group, with 10 mice in each group, and 10 SAMR1 mice as normal group. The acupuncture group received acupuncture at Baihui (GV 20), Xuehai (SP 10), Shenshu (BL 23), and Geshu (BL 17). Xuehai (SP 10), Shenshu (BL 23), and Geshu (BL 17) were stimulated on the left side first and then on the right side alternately, once a day. The non-acupoint group received acupuncture at fixed bilateral non-meridian, non-acupoint points under the ribs, once a day. The model and normal groups underwent equivalent handling and restraint stress without acupuncture. The donepezi group received 1 mg/kg donepezil via gastric gavage daily. All groups were treated for 4 weeks. After treatment, Morris water maze tests (to record orientation sailing latency, number of traverses through the platform quadrant) and open field (to record distance travelled) were used to evaluate learning and memory abilities; hippocampal neuronal damage was analyzed via HE and Nissl staining; mitochondrial membrane potential and reactive oxygen species (ROS) were measured using assay kits; Western blot and qRT-PCR were performed to detect silencing information regulator 1 (SIRT1), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), and mRNA expression levels.
Results
2
Compared with the normal group, mice in the model group and non-acupoint group showed elevated orientation sailing latency and relative multiplicity of ROS in hippocampal tissues, and reduced number of traverses through the platform quadrant, distance of movement in the open-field experiment, number of Nissl-staining-positive cells in the hippocampal tissues, mitochondrial membrane potential, and protein levels and mRNA expression of SIRT1, PGC-1α (
P
<0.01); HE staining showed that the hippocampal tissues of the mice was loosely arranged, with reduced number of neurons and vacuolar degeneration; Nissl staining showed that pyramidal neurons in the hippocampal region were not neatly arranged, and the number of Nissl bodies in the cytoplasm was less and the staining was lighter. Compared with the model group, mice in the acupuncture group and donepezil group had lower orientation saili
ng latency and relative multiplicity of ROS in the hippocampal tissue, higher number of traverses through the platform quadrant, distance of movement in the open-field experiment, number of Nissl-stained positive cells in the hippocampal tissue, mitochondrial membrane potential, and protein levels and mRNA expression of SIRT1 and PGC-1α (
P
<0.01), and HE staining and Nissl staining showed significant improvement in hippocampal histopathological damage. Compared with the donepezil group, the orientation sailing latency shortened in the acupuncture group of mice (
P
<0.01).
Conclusion
2
Kidney-tonifying and blood-activating acupuncture method
can alleviate the SIRT1/PGC-1α signalling pathway in the hippocampal tissue and improve the mitochondrial function, thus alleviating the neuronal damage, which is one of the possible mechanisms for its treatment of Alzheimer's disease.
JIA J , NING Y , CHEN M , et al . Biomarker changes during 20 years preceding Alzheimer's disease [J]. N Engl J Med , 2024 , 390 ( 8 ): 712 - 722 .
TREJO-LOPEZ JA , YACHNIS AT , PROKOP S . Neuropathology of Alzheimer's disease [J]. Neurotherapeutics , 2022 , 19 ( 1 ): 173 - 185 .
NABI SU , KHAN A , SIDDIQUI EM , et al . Mechanisms of mitochondrial malfunction in Alzheimer's disease: new therapeutic hope [J]. Oxid Med Cell Longev , 2022 , 2022 : 4759963 . doi: 10.1155/2022/4759963 http://dx.doi.org/10.1155/2022/4759963 .
DONG W , YAN L , TAN Y , et al . Melatonin improves mitochondrial function by preventing mitochondrial fission in cadmium-induced rat proximal tubular cell injury via SIRT1-PGC-1α pathway activation [J]. Ecotoxicol Environ Saf , 2022 , 242 : 113879 . doi: 10.1016/j.ecoenv.2022. 113879 http://dx.doi.org/10.1016/j.ecoenv.2022.113879 .
ZHAO Y , ZHANG J , ZHENG Y , et al . NAD + improves cognitive function and reduces neuroinflammation by ameliorating mitochondrial damage and decreasing ROS production in chronic cerebral hypoperfusion models through Sirt1/PGC-1α pathway [J]. J Neuroinflammation , 2021 , 18 ( 1 ): 207 .
肖岚 , 董克礼 . 董克礼教授运用补肾活血法治疗阿尔茨海默病经验 [J]. 中华中医药杂志 , 2015 , 30 ( 2 ): 435 - 437 .
朱宏 , 董克礼 , 吴岳 , 等 . 补肾活血法对阿尔茨海默病患者认知功能改善的影响 [J]. 中国老年学杂志 , 2010 , 30 ( 11 ): 1493 - 1495 .
萨依腊西·杰恩斯努尔 , 肖岚 , 龙见汝 , 等 . 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 [J]. 中国中医药信息杂志 , 2018 , 25 ( 9 ): 35 - 38 .
梁梅亭 , 朱宏 , 董克礼 , 等 . 补肾活血针刺法对SAMP8小鼠杏仁核蛋白质组学表达的影响 [J]. 中国中医药信息杂志 , 2018 , 25 ( 3 ): 58 - 63 .
中国针灸学会 . 实验动物常用穴位名称与定位 第3部分:小鼠 [J]. 针刺研究 , 2021 , 46 ( 5 ): 445 - 446 .
卞金玲 , 张春红 . 石学敏院士针刺手法量学的概念及核心 [J]. 中国针灸 , 2003 , 23 ( 5 ): 38 - 40 .
张利达 , 高永龙 , 蔡兴慧 , 等 . 近5年针刺保护神经作用研究概况 [J]. 陕西中医学院学报 , 2017 , 40 ( 4 ): 137 - 139 .
朱宏 , 董克礼 , 吴岳 , 等 . 针刺对阿尔茨海默病患者异构前列腺素的影响 [J]. 中国针灸 , 2010 , 30 ( 1 ): 18 - 21 .
高旭红 , 赵海花 , 赖红 , 等 . 多奈哌齐对AD大鼠海马结构神经元的保护作用 [J]. 中国组织化学与细胞化学杂志 , 2012 , 21 ( 1 ): 33 - 36 .
刘欣媛 , 杜艳军 , 王丽 , 等 . 电针抗炎效应在SAMP8小鼠神经元损伤中的促保护作用 [J]. 针刺研究 , 2023 , 48 ( 2 ): 133 - 138 .
金艳 , 李阳 , 雷旭伟 . 多奈哌齐治疗AD源性轻度认知功能障碍患者的临床研究 [J]. 中国临床药理学杂志 , 2024 , 40 ( 18 ): 2616 - 2620 .
ALEXIOU A , SOURSOU G , CHATZICHRONIS S , et al . Role of GTPases in the regulation of mitochondrial dyna-mics in Alzheimer's disease and CNS-related disorders [J]. Mol Neurobiol , 2019 , 56 ( 6 ): 4530 - 4538 .
AGRAWAL I , JHA S . Mitochondrial dysfunction and Alzheimer's disease: role of microglia [J]. Front Aging Neurosci , 2020 , 12 : 252 . doi: 10.3389/fnagi.2020. 00252 http://dx.doi.org/10.3389/fnagi.2020.00252 .
LITWINIUK A , BARANOWSKA-BIK A , DOMAŃSKA A , et al . Contribution of mitochondrial dysfunction combined with NLRP3 inflammasome activation in selected neurodegenerative diseases [J]. Pharmaceuticals (Basel) , 2021 , 14 ( 12 ): 1221 .
YE CY , LEI Y , TANG XC , et al . Donepezil attenuates Aβ-associated mitochondrial dysfunction and reduces mitochondrial Aβ accumulation in vivo and in vitro [J]. Neuropharmacology , 2015 , 95 : 29 - 36 . doi: 10.1016/j.neuropharm.2015.02.020 http://dx.doi.org/10.1016/j.neuropharm.2015.02.020 .
LIANG D , ZHUO Y , GUO Z , et al . SIRT1/PGC-1 pathway activation triggers autophagy/mitophagy and attenuates oxidative damage in intestinal epithelial cells [J]. Biochimie , 2020 , 170 : 10 - 20 . doi: 10.1016/j.biochi. 2019.12.001 http://dx.doi.org/10.1016/j.biochi.2019.12.001 .
YIN Z , GAO D , DU K , et al . Rhein ameliorates cognitive impairment in an APP/PS1 transgenic mouse model of Alzheimer's disease by relieving oxidative stress through activating the SIRT1/PGC-1α pathway [J]. Oxid Med Cellul Longev , 2022 , 2022 : 2524832 . doi: 10.1155/2022/2524832 http://dx.doi.org/10.1155/2022/2524832 .
ZHU C , ZHANG Z , ZHU Y , et al . Study on the role of Dihuang Yinzi in regulating the AMPK/SIRT1/PGC-1α pathway to promote mitochondrial biogenesis and improve Alzheimer's disease [J]. J Ethnopharmacol , 2025 , 337 ( Pt 2 ): 118859 .
0
浏览量
0
下载量
1
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010102007558号