人参皂苷Ro延长秀丽隐杆线虫的寿命并增强氧化应激抵抗力

Ginsenoside Ro Prolongs Longevity and Enhances Stress Resistance in Caenorhabditis elegans

目的以秀丽隐杆线虫((Caenorhabditis elegans,C.elegans),下文简称线虫)及其突变株为生物模型,研究人参皂苷Ro(Ginsenoside-Ro,G-Ro)对健康寿命的影响及抗氧化作用。方法实验设空白对照组、不同剂量G-Ro(5、10、20μmol·L^(-1))药物组、阳性药物对照组(40μmol·L^(-1)二甲双胍),研究G-Ro对线虫寿命、脂褐素荧光的影响。另外,在上述基础上利用10 mmol·L^(-1)百草枯(Paraquat,PQ)建立氧化损伤模型,探究G-Ro对线虫氧化应激状态下寿命及活性氧自由基(Reactive oxygen species,ROS)的影响。利用绿色荧光蛋白标记的DAF-16::GFP、SOD-3::GFP线虫及突变株daf-16(mu86)分析G-Ro对DAF-16及SOD-3的调节作用。结果与空白对照组相比,20μmol·L^(-1) G-Ro延长了线虫寿命(***P<0.001);在氧化应激模型中,延长线虫寿命(#P<0.05);降低脂褐素荧光水平(11.70%)。20μmol·L^(-1) G-Ro降低线虫内源性ROS(11.92%,*P<0.05),降低氧化应激后升高的ROS(79.67%,*P<0.001)。与空白对照组相比,20μmol·L^(-1) GRo提高叉头转录因子(Fork head box O,FOXO)同系物DAF-16去磷酸化入核率(51.00%);增强SOD-3荧光强度(19.85%,***P<0.001)。在正常及氧化应激状态下,G-Ro对突变体daf-16(mu86)寿命的影响均无统计学意义。结论在正常及氧化应激状态下G-Ro延长线虫寿命的作用,可能与其上调SOD-3,清除累积的ROS相关。DAF-16的转录活性可能介导G-Ro效应。

Objective To study the effects of Ginsenoside Ro(G-RO)on health life and antioxidant activity of Caenorhabditis elegans(C.elegans)and its mutant strains as biological models.Methods The blank control group,different doses of G-Ro(5,10,20μmol·L^(-1))groups and positive control group(40μmol·L^(-1) metformin)were set up to study the effects of G-Ro on the lifespan and lipofuscin fluorescence of C.elegans.The oxidative damage model was established by using 10 mmol·L^(-1) paraquat(PQ)to investigate the effects of G-Ro on Reactive oxygen species(ROS)in C.elegans.We used DAF-16::GFP,SOD-3::GFP,and mutant daf-16(mu86)to explore the mechanism of G-Ro.Results Compared with the blank control group,20μmol·L^(-1) G-Ro prolonged the lifespan of C.elegans(***P<0.001).In the oxidative stress model constructed by PQ,the lifespan of nematode was prolonged(#P<0.05).The fluorescence level of lipofuscin decreased by 11.70%.20μmol·L^(-1) G-Ro decreased ROS levels(11.92%,*P<0.05),decreased ROS levels after oxidative stress induced by 10 mmol·L^(-1) PQ(79.67%,*P<0.001),increased Fork head box O(FOXO)homologous Daf-16 dephosphorylation into nucleus rate(51.00%),increased the fluorescence intensity of SOD-3(19.85%,***P<0.001).However,the effect of G-Ro on the lifespan of mutant daf-16(mu86)was not statistically significant under normal and oxidative stress conditions.Conclusion The effect of G-Ro on prolonging life span of nematodes under normal and oxidative stress conditions may be related to up-regulation of SOD-3 and clearance of accumulated ROS.Transcriptional activity of DAF-16 may mediate G-Ro effect.