Study Of Anti-aging Effects Of Ginsenoside From Panax Ginseng In Caenorhabditis Elegans

Aging is inevitable,but with the improvement of people’s living standard,people pay more attention to health preservation and advocate healthy aging.Panax ginseng is a traditional Chinese medicine.For many years,the anti-aging effect of Panax ginseng and its components have been one of the hot topics of research,but there are few studies to explore the anti-aging effect and mechanism of Panax ginseng with whole animals.In this study,Caenorhabditis elegans（C.elegans）,was used as a whole animal model to analyze the effects of ginseng total saponins（GTS）and four kinds of ginsenosides on the overall lifespan of C.elegans,and to analyze the possible mechanism of action.The main research results are as follows:GTS（1,10,100μg/m L）could significantly prolong the mean lifespan of C.elegans by 8.17±1.60%,9.11±2.04%and 7.64±1.28%,respectively,rather than altering the growth and metabolism of E.coli OP50.We futher studied the change of C.elegans markers of aging after GTS-treated,and found that GTS have no obvious toxic effect on reproduction,body length,locomotion ability and pharyngeal pumping.Moreover,GST could reduce fat and lipofuscin accumulation to promote healthspan.GTS could significantly improve the survival ability of C.elegans under heat stress and oxidative stress,which indicated that GTS could reduce the ROS level of C.elegans under stress conditions.These results are consistent with the widely recognized antioxidant effect of GTS,which may be related to its anti-aging effect.To further understand the components of GTS,we analyzed the components of GTS by HPLC.We found that GTS mainly included four kinds of ginsenosides,Rg₁,Re,Rg₂ and Rd.1μg/m L ginsenoside Rd had the similar effect of delaying the aging of C.elegans,and could prolong the mean lifespan of C.elegans by 13.26±2.25%,while 1μg/m L Re,Rg₁ and Rg₂ did not prolong the lifespan of C.elegans.To further investigate the molecular mechanism by which GTS prolong the lifespan of C.elegans,we systematically analyzed the changes of transcriptome genes by RNA-Seq,and found that 26 genes were significantly up-regulated and 10 genes were significantly down-regulated.First,we found that up-regulated（C56C10.7,F36H5.14,gnrr-1,nspc-2,coa-4）and down-regulated（F35E8.13,atx-3,wdr-23,Y102A11A.7,cat-2）were the most significant differentially expressed genes,which regulated by daf-16,skn-1,hsf-1（Insulin/IGF-1 signaling pathway）and nuclear receptor genes daf-12.Secondly,according to the KEGG pathway suggested that GTS regulated JNK-1,PHI-62 and NHR-80 to exert its antiaging effect in C.elegans.JNK-1 and PHI-62were located in the upstream of DAF-16,which could activate different functions mediated by DAF-16 and jointly promote the longevity of C.elegans.Therefor,we focus on the m RNA levels of daf-16,skn-1,hsf-1,daf-12 and nhr-80.The effects of GTS-treated on the lifespan of daf-16,hsf-1 and skn-1 mutant strains were investigated.It was found that GTS did not prolong lifespan when daf-16and hsf-1 were deficient,suggesting that the anti-aging effect of GTS may be related to daf-16 and hsf-1,and simultaneously activate DAF-16 nuclear translocations.Furthermore,q RT-PCR result was found that the m RNA levels of daf-16,hsf-1 and their downstream related genes sod-3 and hsp-16.2 were significantly increased.It was also found that GTS up-regulated the m RNA levels of daf-12,nhr-80 and their downstream genes fard-1 and fat-6.The results showed that the anti-aging effect of GTS was related to daf-16,hsf-1,daf-12 and nhr-80.Next,we studied whether the molecular mechanism of ginsenoside Rd to prolong the lifespan of C.elegans was similar to GTS.Through q RT-PCR analysis,we verified that ginsenoside Rd can also up-regulate the m RNA expression levels of daf-16,daf-12,nhr-80 and their downstream related genes sod-3,fard-1 and fat-6,and activate the nuclear translocations of DAF-16.These results indicated that the main component of GTS in prolonging the lifespan of C.elegans was ginsenoside Rd.In conclusion,by adding GTS or ginsenoside Rd to the diet of C.elegans,we confirmed that both GTS and ginsenoside Rd can delay aging.At the gene level,it was found that GTS and ginsenoside Rd had a common mechanism of action,which could delay aging by increasing the expressions of daf-16,hsf-1,daf-12,nhr-80 and their downstream genes,and promoted the nuclear translocation of DAF-16.The anti-aging mechanism of ginsenoside Rd is worthy of further study,and these findings will help us to further understand the anti-aging effect and mechanism of Panax ginseng in whole model organisms.