红景天苷对海马神经元自然衰老的影响和机制研究

中华养生保健 ›› 2024, Vol. 42 ›› Issue (8): 16-20.

红景天苷对海马神经元自然衰老的影响和机制研究

徐新颖^1,2, 朱琳², 刘姿杉², 倪钦帅², 于竹芹^2,3,*

1.济南市济阳区中医院ICU科,山东济南,251400;
2.青岛大学医学部中西医结合中心,山东青岛,266021;
3.青岛市第六人民医院内科,山东青岛,266023

出版日期:2024-04-16 发布日期:2024-04-08
通讯作者: ^* 于竹芹,E-mail：yuzhuq2008@163.com。
作者简介:徐新颖（1983—）,女,汉族,籍贯：山东省济南市,硕士研究生,主治医师,研究方向：中西医结合临床。
基金资助:
山东省中医药科技项目（M-2022004）; 青岛市中医药科研计划项目（2020-ZYZ003）

The Effect and Mechanism of Salidroside on Natural Senescence of Hippocampal Neurons

XU Xin-ying^1,2, ZHU Lin², LIU Zi-shan², NI Qin-shuai², YU Zhu-qin^2,3,*

1. Department of ICU, Jiyang District TCM Hospital, Jinan Shandong 251400, China;
2. Institute of Integrative Medicine, Qingdao University Medical College, Qingdao Shandong 266021, China;
3. Department of Medicine, The Sixth Peoples'Hospital of Qingdao, Qingdao Shandong 266023, China

Online:2024-04-16 Published:2024-04-08

摘要/Abstract

摘要： 目的探讨红景天苷延缓原代海马神经元自然衰老的作用及可能的机制。方法分离并培养大鼠原代海马神经元至自然衰老,随机分为短期培养（ST）组、长期培养（LT）组、红景天苷低和高浓度（10 µM和20 µM）组、阳性药环黄芪醇（5 µM）组,使用细胞计数法检测神经元存活率,使用衰老β-半乳糖苷酶（SA-β-gal）染色检测神经元衰老,使用流式细胞术检测神经元凋亡,使用Western blot检测端粒酶逆转录酶（TERT）表达,使用PCR-ELISA法检测端粒酶活性。结果与ST组比较,LT组的神经元出现神经元退化,胞体萎缩,细胞数量和存活率显著降低（P<0.01）,神经元SA-β-gal阳性率和凋亡率明显增加（P<0.01）,而神经元内端粒酶和TERT表达水平显著降低（P<0.01）;给予红景天苷和环黄芪醇处理后,与LT组神经元相比,细胞存活率明显提高（P<0.05）,细胞SA-β-gal阳性率显著降低（P<0.05）,神经元凋亡率明显减少（P<0.01）,神经元内的端粒酶和TERT表达水平显著提高（P<0.01）。结论红景天苷能够减少海马神经元凋亡,提高端粒酶的表达水平,进而延缓神经元衰老。

关键词: 红景天苷, 原代培养, 海马神经元, 细胞凋亡, 端粒酶

Abstract: Objective To investigate the effect and mechanism of salidroside delaying natural aging of primary hippocampal neurons. Methods Rat hippocampal neurons were isolated and then cultured into aging neurons, which were randomly divided into short-term (ST) group, long-term (LT) group, salidroside low- and high-concentration (10 µM and 20 µM) groups, positive drug cycloastragenol group (5 µM). Cell count method (CCK-8) was used to detect hippocampal neurons survival rate, senescence-associated β-galactosidase (SA-β-gal) staining kit was used to detect the hippocampus neuron aging, the apoptotic rate of hippocampal neurons was detected by flow cytometry, the expression of telomerase reverse transcriptase (TERT) was detected by western blot, PCR-ELISA method was used to detect telomerase activity of hippocampal neurons. Results Compared with the ST group, the hippocampal neurons appeared neuron degeneration, cell body contraction, and the cell survival rate decreased significantly (P<0.01), while SA-β-gal positive rate increased significantly in LT group (P<0.01). The apoptotic rate of neurons was significantly increased (P<0.01), while telomerase activities and TERT expression of neurons decreased obviously (P<0.01). Compared with LT group, the neuronal survival rate treated with salidroside and cycloastragenol were significantly increased (P<0.05), while SA-β-gal positive rate and apoptotic rate of neurons was significantly decreased (P<0.05), and the telomerase activities and TERT expression were significantly increased (P<0.01). Conclusion Salidroside could reduce the neuronal apoptosis and increase the telomerase activity to delay the senescence of hippocampal neurons.

Key words: salidroside, primary culture, hippocampal neurons, apoptosis, telomerase

中图分类号:

R453.9

徐新颖, 朱琳, 刘姿杉, 倪钦帅, 于竹芹. 红景天苷对海马神经元自然衰老的影响和机制研究[J]. 中华养生保健, 2024, 42(8): 16-20.

XU Xin-ying, ZHU Lin, LIU Zi-shan, NI Qin-shuai, YU Zhu-qin. The Effect and Mechanism of Salidroside on Natural Senescence of Hippocampal Neurons[J]. ZHONGHUA YANGSHENG BAOJIAN, 2024, 42(8): 16-20.

参考文献

[1] HE S, SHARPLESS N E.Senescence in health and disease[J]. Cell,2017, 169(6):1000-1011.
[2] CALVO-RODRÍGUEZ M, DE LA FUENTE C, GARCÍA-DURILLO M, et al. Aging and amyloid β oligomers enhance TLR4 expression, LPS-induced Ca2+ responses, and neuron cell death in cultured rat hippocampal neurons[J]. J Neuroinflammation,2017,14(1):24-33.
[3] WHITTEMORE K, DEREVYANKO A, MARTINEZ P, et al.Telomerase gene therapy ameliorates the effects of neurodegeneration associated to short telomeres in mice[J]. Aging (Albany NY),2019,11(10):2916-2948.
[4] 冀小伟,张连城.中医对衰老的认识[J].中医杂志,2013,54(17):1527-1529.
[5] 孙安琪,颜天华,巨修练.红景天苷药理作用及分子机制的研究进展[J].时珍国医国药,2018,29(6):1440-1443.
[6] 于冬建. 红景天药理学研究及临床作用新进展[J].中医临床研究,2020, 12(18):136-138.
[7] 倪钦帅,杨光文,王悦,等.红景天苷对C57衰老小鼠的肝保护作用研究[J].世界中西医结合杂志,2023,18(7):1334-1340.
[8] 杨光文,倪钦帅,朱琳,等.红景天苷通过上调端粒酶表达发挥延缓小鼠脑衰老的作用机制[J].世界中西医结合杂志,2023,19(9):1750-1755.
[9] ZHUANG W, YUE L F, DANG X F, et al.Rosenroot (rhodiola): potential applications in aging-related diseases[J]. Aging Dis,2019,10(1):134-146.
[10] CALVO M, SANZ-BLASCO S, CABALLERO E, et al.Susceptibility to excitotoxicity in aged hippocampal cultures and neuroprotection by non-steroidal anti-inflammatory drugs: role of mitochondrial calcium[J]. J Neurochem,2015,132(4):403-417.
[11] 林晓月,孙新.红景天抗衰老作用机制研究进展[J].中国老年学杂志,2018,38(13):3299-3300.
[12] LI T, ZHANG W, KANG X, et al.Salidroside protects dopaminergic neurons by regulating the mitochondrial MEF2D-ND6 pathway in the MPTP/MPP+-induced model of Parkinson’s disease[J]. J Neurochem,2020,153(2):276-289.
[13] WANG H, LI Q, SUN S, et al.Neuroprotective effects of salidroside in a mouse model of Alzheimer’s disease[J]. Cell Mol Neurobiol,2020, 40(7):1133-1142.
[14] 张霜梅,祝维峰,王安荣,等.红景天苷通过Nrf2-HO-1保护Aβ25-35诱导的原代皮层神经元损伤[J].中华中医药学刊,2021,39(1):101-106,277-282.
[15] WU Y, WANG Y, WU Y, et al.Salidroside shows anticonvulsant and neuroprotective effects by activating the Nrf2-ARE pathway in a pentylenetetrazol-kindling epileptic model[J]. Brain Res Bull,2020,164(1):14-20.
[16] SODERO A O, WEISSMANN C, LEDESMA M D, et al.Cellular stress from excitatory neuro- transmission contributes to cholesterol loss in hippocampal neurons aging in vitro[J]. Neurobiol Aging,2011,32(6):1043-1053.
[17] MAO G X, XU X G, WANG S Y, et al.Salidroside delays cellular senescence by stimulating mitochondrial biogenesis partly through a miR-22/SIRT-1 pathway[J]. Oxid Med Cell Longev,2019:5276096.
[18] ZHANG L, DING W, SUN H, et al.Salidroside protects PC12 cells from MPP+-induced apoptosis via activation of the PI3K/Akt pathway[J]. Food Chem Toxicol,2012,50(8):2591-2597.
[19] LI M, LI S C, DOU B K, et al.Cycloastragenol upregulates SIRT1 expression, attenuates apoptosis and suppresses neuroinflammation after brain ischemia[J]. Acta Pharmacol Sin,2020,41(8):1025-1032.
[20] HARLEY C B, FUTCHER A B, GREIDER C W.Telomeres shorten during aging of human fibroblasts[J]. Nature,1990,345(6274):458-460.
[21] LEE J, JO Y S, SUNG Y H, et al.Telomerase deficiency affects normal brain functions in mice[J]. Neurochem Res,2010,35(2):211-218.
[22] JASKELIOFF M, MULLER F L, PAIK J H, et al.Telomerase reactivation reverses tissue degeneration in aged telomerase-deficient mice[J]. Nature,2011,469(7328):102-106.