山茱萸多糖延缓秀丽隐杆线虫衰老的药效评价及机制研究

Pharmacodynamic evaluation and mechanism study of Corni Fructus polysaccharide on anti-aging of Caenorhabditis elegans

中文关键词:  山茱萸多糖  秀丽隐杆线虫  寿命  抗衰老  抗氧化

英文关键词:Corni Fructus polysaccharides  Caenorhabditis elegans  lifespan  anti-aging  anti-oxidant

中文摘要:

      目的 考察山茱萸多糖(Corni Fructus polysaccharides, CFP)对秀丽隐杆线虫衰老的延缓作用并探讨其作用机制。方法 以野生型秀丽隐杆线虫(N2线虫)作为动物模型, 利用百草枯建立氧化应激模型, 筛选CFP的最佳药效浓度; 测定CFP对N2线虫的急性热应激、寿命、生殖及咽泵运动能力的影响; 检测并比较给药组与空白组活性氧簇(reactive oxygen species, ROS)及脂褐素水平; 运用实时荧光定量聚合酶链式反应(polymerase chain reaction, PCR)检测CFP对N2线虫衰老相关基因mRNA表达的影响。结果 通过抗氧化应激实验测得CFP最佳给药浓度为150 μg/mL; 急性热应激实验中, CFP给药组较空白组的生存曲线右移; 不同浓度CFP给药组较空白组的存活率更高(P<0.01), 平均寿命(P<0.001)和最长寿命均延长; 给药组子代数量在第1、3、5、7 d无显著性差异(P>0.05); 50、150 μg/mL的CFP组均可增强N2线虫咽泵运动能力(P<0.05); 不同浓度CFP给药均能显著降低N2线虫体内ROS含量和减少脂褐素沉积; 与空白组相比, 150 μg/mL CFP给药5 d后, DAF-16、SKN-1、SOD-3及CTL-2的mRNA转录水平均显著上调, AGE-1的mRNA转录水平显著下调。结论 CFP能在一定程度上延缓秀丽线虫的衰老, 其作用机制与提高线虫体内抗氧化应激能力、抑制ROS产生、减少脂褐素的沉积, 以及调控胰岛素/胰岛素样生长因子(insulin like growth factor, IGF)-1信号(insulin/IGF-1 signaling, IIS)通路上的基因表达有关。

英文摘要:

      Objective To investigate the anti-aging effects of Corni Fructus polysaccharides (CFP) on Caenorhabditis elegans and explore its mechanism. Methods The wild-type C. elegans (N2) was used as an animal model, and the oxidative stress model was established by paraquat to screen the optimal pharmacodynamic concentration of CFP. The effects of CFP on acute heat stress, life span, reproduction and pharyngeal pumping ability of N2 nematode were determined. Reactive oxygen species (ROS) and lipofuscin levels were detected and compared between the administration group and the blank control group. The real-time fluorescent quantitative polymerase chain reaction (PCR) was used to examine the effect of CFP on mRNA expression of aging-related genes in N2 nematodes. Results The optimal concentration of CFP was 150 μg/mL through the anti-oxidative stress test; in the acute heat stress experiment, the survival curve of the CFSP administration group was shifted to the right as compared with that of the blank control group, and the survival rate of CFP groups administered with different concentrations was higher than that of the blank control group (P<0.01), and the average lifespan (P<0.001) and the longest lifespan were both prolonged. There was no significant difference in the number of offspring on the 1st, 3rd, 5th and 7th d in the administration group (P>0.05); the CFP groups (50 μg/mL or 150 μg/mL) could both enhance the pharyngeal pump locomotor ability of N2 nematodes (P<0.05); CFP at different concentrations could significantly reduce ROS content and lipofuscin deposition in N2 nematodes; compared with the blank group, the mRNA transcription levels of DAF-16, SKN-1, SOD-3, and CTL-2 were significantly up-regulated and that of AGE-1 was significantly down-regulated after 5 days of administration of 150 μg/mL CFP. Conclusions CFP can delay the aging of C. elegans to some extent, and its mechanism is related to improving the anti-oxidative stress ability of C. elegans, inhibiting the production of ROS, reducing the deposition of lipofuscin, and regulating the gene expression on insulin/insulin like growth factor (IGF)-1 signaling (IIS) pathway.