| Aging and related diseases have become a widespread concern in the context of the increasing global aging situation.Aging is influenced by many factors,among which the main theory of aging is based on the free radical theory.It is believed that along with the aging process,the free radicals in the human body will accumulate too much,thus causing damage to cell membranes,nucleic acids,proteins and various enzymes,thus causing serious damage to cellular functions and leading to dysfunction of tissues and organs,resulting in the slow aging of the body.In recent years,with the research on delaying aging,it has been found that a long and healthy life is far more important than simply extending life expectancy.Prolonging the body’s tolerance to stress and reducing the occurrence of aging diseases is an effective way to fight aging.Longevity is an important way to achieve good health and improve quality of life in old age.And the current way for delaying aging is to give medication.Ginseng fruit,one of the non-medicinal parts of ginseng,is rich in ginsenosides and has similar pharmacological activities as ginseng.It has been proven that total ginsenosides(GBS)have protective effects on cardiovascular system,endocrine system,immune system and anti-aging.Ginseng berries are abundant,but their utilization is low and has a broad research potential,while GBS is widely used for its pharmacological activities such as anticancer,antitumor,antidiabetic and anti-aging.The short life cycle and clear genetic background associated with lifespan of Cryptobacterium hidradenum are well suited for pharmacological evaluation and mechanism study of anti-aging drugs.In this study,we investigated the effect and mechanism of GBS in anti-aging by using Cryptobacterium hidradenum as an experimental subject.1.Analysis of ginsenoside composition and percentage of GBS extracted from Ginseng fruit by HPLCIn this study,ginseng fruits were crushed and eluted with water and 70% ethanol on a macroporous adsorbent resin column to obtain GBS,and then the extracted GBS was analyzed by HPLC.The analysis revealed that GBS was rich in 18 ginsenosides,namely Rg1,Re,Rb1,Rc,Rg2,Rh1,Rb2,Rb3,Rd,Rg6,F4,Rk3,Rh4,Rg3,PPT,Rk1,Rg5,R-Rh2.14.18%,and 13.55%,respectively.2.Study on the anti-aging effect and mechanism of GBS on Caenorhabditis elegansIn this study,different concentrations of GBS(0.125 mg/m L,0.25 mg/m L,0.5 mg/m L,1mg/m L 2 mg/m L,4 mg/m L)were administered to wild-type N2 nematodes in lifespan experiments,so as to screen the most effective GBS with lifespan extension.subsequent experiments will be conducted using this concentration.It was found that 0.25 mg/m L of GBS was the concentration with the most lifespan-prolonging effect.In the experiments on bacterial growth rate,food preference assay and swallowing ability,it was found that GBS had no effect on OP50 and that GBS did not act as a dietary restriction and nematode avoidance leading to life extension,thus confirming that GBS acts directly on nematodes and not indirectly on other aspects to produce life extension results.In reproduction experiments,it was also confirmed that GBS administration did not affect nematode reproduction and that the life-extending effect was not produced by inhibition of reproduction.In motility assays,GBS was also found to increase nematode motility on days 10 and 15.In the heat stress and oxidative stress assays,it was also found that GBS increased the nematode’s resistance to stress,thereby extending the nematode’s lifespan under stress.In the lipofuscin experiment,GBS was also found to be effective in reducing the expression of lipofuscin produced in nematodes on days 10 and 15.In the ROS assay,GBS was found to be effective in reducing ROS production in nematodes.GBS was also found to be unable to extend the lifespan of mutant nematodes CB1370,CF1038,EU31,EU1,and VC199 in lifespan experiments,so the effect of GBS with lifespan extension was dependent on DAF-2,DAF-16,Sir-2.1,and SKN-1.In GFP nematodes TJ356(DAF-16∷GFP),CF1553(SOD-3∷GFP),TJ375(HSP-16.2∷GFP)and CL2166(GST-4∷GFP),GBS was found to increase the expression of the genes bound by the above GFPs,so it can be demonstrated that nematodes with lifeextending effects are dependent on DAF-16,SOD-3,HSP-16.2 and GST-4.Finally,by Finally,the real-time quantitative PCR experiments revealed that GBS could decrease the expression of DAF-2 and increase the expression of DAF-16,SKN-1,Sir-2.1,SOD-3,GST-4 and HSP-16.2.3.Network pharmacology-based analysis of the anti-aging mechanism of action of GBSA total of 18 ginsenosides in GBS were analyzed using HPLC.The results of network pharmacology prediction showed that there were 440 common targets of the 18 active ingredients and aging in GBS,including 17 key targets,and the top 5 key targets were screened as HSP90AA1,PI3 CA,PTAFR,STAT3,IL2.The results of GO analysis and KEGG signaling pathway enrichment analysis showed that GBS anti-aging mainly acted on PI3K-Akt,insulin signaling pathway,etc.insulin signaling pathway,etc.The molecular docking results showed that the active components of GBS,Rk1、Rb2 and Rc,had good binding activities to the key targets.In conclusion,this paper makes full use of the Caenorhabditis elegans model,in which the pharmacological activities of GBS are represented in different states and models of the nematode to provide a material basis for subsequent studies,as well as to provide research ideas for the nonmedicinal parts of ginseng. |
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