基于秀丽隐杆线虫模型的林下山参抗衰老作用研究 ^*

Study on the anti-aging effects of mountain-cultivated ginseng based on the Caenorhabditis elegans model

摘要

目的 采用秀丽隐杆线虫模型评价林下山参延缓衰老的作用, 并初步探究其作用机制。方法 以秀丽隐杆线虫为衰老模式动物模型, 通过热应激实验, 观察林下山参水提物对 N2 野生型秀丽隐杆线虫耐热性的影响, 同时检测线虫寿命、体长、运动能力、超氧化物歧化酶 (SOD) 及丙二醛 (MDA) 含量、脂褐素水平及活性氧 (ROS) 水平等衰老相关生理指标的变化。通过网络药理学, 进行 GO 功能富集分析及 KEGG 通路富集分析, 并构建 “林下山参—化学成分—交集靶点—衰老—信号通路” 网络。最后通过实时荧光定量聚合酶链反应 (RT-qPCR) 对潜在信号通路进行初步验证。结果 林下山参水提物可有效延长秀丽隐杆线虫寿命、提高热应激后的存活率、增加体长、增强运动能力、增加 SOD 含量; 并减少 MDA、脂褐素和 ROS 等有害物质的积累, 差异均具有统计学意义 (P<0.05)。网络药理学分析表明, 生物学过程中富集靶点较多; 林下山参可能通过调节 PI3K-AKT、Ras 及 HIF-1 等多条信号通路发挥抗衰老的药理作用。此外, RT-qPCR 结果表明, 林下山参可明显抑制线虫体内 PI3K-AKT 信号通路相关 age-1、akt-1、daf-2 等 mRNA 的表达, 同时显著提高 daf-16 的 mRNA 表达水平。结论 林下山参可有效延缓秀丽隐杆线虫衰老, 可能与抑制 PI3K-AKT 信号通路密切相关。

Abstract

Objective To evaluate the anti-aging effects of mountain-cultivated ginseng using the Caenorhabditis elegans model and preliminarily investigate its mechanisms through network pharmacology analysis and RT-qPCR experiments. Methods The study employed Caenorhabditis elegans as an aging model organism. The impact of the water extract of mountain-cultivated ginseng on the thermotolerance of N2 wild-type Caenorhabditis elegans was observed using a heat stress challenge assay. Aging-related physiological indicators were measured, including lifespan, body length, locomotor ability, superoxide dismutase (SOD) and malondialdehyde (MDA) levels, lipofuscin accumulation, and reactive oxygen species (ROS) levels. Gene Ontology functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed based on network pharmacology. The “mountain-cultivated ginseng-chemical components-intersection targets-aging-signaling pathways” network was construct, followed by preliminary validation of potential signaling pathways via uantitative real-time polymerase chain reaction (RT-qPCR). Results The water extract of mountain-cultivated Ginseng significantly extended Caenorhabditis elegans lifespan, increased survival rates under heat stress, enhanced body length and locomotor activity, elevated SOD levels, and reduced accumulations of MDA, lipofuscin, and ROS, and the differences were statistically significant (P<0.05). Network pharmacological analysis indicates that biological processes exhibit a significant enrichment of targets; mountain-cultivated ginseng may exert its anti-aging pharmacological effects by modulating multiple signaling pathways, including PI3K-AKT, Ras, and HIF-1 signaling pathways. RT-qPCR results confirmed mountain-cultivated ginseng significantly downregulated the mRNA expression of age-1, akt-1, and daf-2, while markedly upregulating daf-16 mRNA expression levels (genes associated with the PI3K-AKT signaling pathway in Caenorhabditis elegans). Conclusion Mountain-cultivated ginseng effectively delays aging in Caenorhabditis elegans, primarily through suppression of the PI3K-AKT signaling pathway.

关键词

林下山参 / 抗衰老 / 秀丽隐杆线虫 / 网络药理学

Key words

Mountain-cultivated ginseng / Anti-aging / Caenorhabditis elegans / Network pharmacology

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基金资助

^*吉林省科技发展计划科技创新基地 (平台) 建设—吉林省科技创新中心项目(YDZJ202402035CXJD)