| At present,the phenomenon of population aging has already become a global problem,and the increasing proportion of the elderly in the population has brought great challenges to the medical system of various countries.To solve this problem,many researchers have devoted themselves to finding drugs that can delay aging and treat related diseases.Caenorhabditis elegans is an ideal model for anti-aging research.It is easy to cultivate,low cost,and has 60%-80%genes homologous with human.Wikstroemia Chamaedaphne Meissn is a traditional Chinese medicinal material rich in diterpenoids and flavonoids,which has antioxidant and cardiovascular disease treatment functions.However,there is no research report on the effect of Wikstroemia Chamaedaphne Meissn on delaying aging.In this paper,the antioxidant activity of total flavonoids extracted from Wikstroemia Chamaedaphne Meissn in vitro is studied and the effect of total flavonoid from Wikstroemia Chamaedaphne Meissn（WCM）on the lifespan and physiological index of nematodes is flavonoid as a model.The molecular mechanism of WCM prolonging the lifespan of nematoides was studied by qRT-PCR,the lifespan of mutant and laser confocal detection.The main research contents and results are as follows:（1）Through the determination of Fe3+reducing power,DPPH removal rate,ABTS.+the scavenging experiment proved that WCM had good antioxidant activity in vitro.（2）The effects of WCM on aging of N2 nematodes were studied by life span experiment.The results showed that 5μg/m L,10μg/m L and 20μg/m L WCM could prolong the life span of N2 nematodes in concentration-dependent manner.Among them,20μg/m L WCM had the best effect,and the prolonging rate was 23.8%（p<0.01）.Quercetin,a positive drug,showed an effect of prolonging the life span of nematodes at50μg/m L,and the prolonging rate was 20.8%（p<0.05）,indicating that WCM has a better anti-aging effect than quercetin.Compared with quercetin,WCM has the advantages of lower concentration and stronger drug effect.Then 20μg/m L WCM group was used as the control group,the life span experiments of 8 compounds in WCM were carried out after conversion of contents.It was found that these 8 flavonoids compounds could prolong the life span of C.elegans to a certain extent,but the effect of WCM total flavonoids was better than that of single compounds,indicating that the components of WCM may prolong the lifespan of C.elegans through synergistic interaction.（3）Through the study of locomotion ability,reproductive ability,development and brown lipid content of C.elegans,it was found that WCM had no adverse effect on reproductive ability and development of C.elegans,and could enhance locomotion ability and reduce brown lipid content of C.elegans,indicating that WCM could prolong the healthy life of C.elegans.（4）Through stress experiments,the contents of ROS and antioxidant enzymes in WCM-treated nematodes and ROS and antioxidant enzymes in WCM-treated nematodes under stress state were studied.It was found that WCM could improve the survival of nematodes under oxidative environment,and it was found that no matter in normal or oxidative stress environment,the antioxidant enzyme activity was significantly higher than that in the control group.WCM can eliminate excess free radicals by enhancing the activity of antioxidant enzymes in nematodes.（5）The molecular mechanism of WCM prolonging C.elegans lifespan was studied by qRT-PCR and mutant lifespan,and it was found that WCM prolonging C.elegans lifespan was independent of dietary restriction pathway.WCM prolongs the lifespan of C.elegans by inhibiting IIS pathway,and its molecular mechanism may be through regulation of daf-16 and skn-1 genes,activation of autophagy genes bec-1 and lgg-1,and removal of damaged organelles and protein macromolecules to prolong the lifespan of C.elegans.In conclusion,WCM can prolong the life span and delay the aging of C.elegans.Its mechanism is related to IIS pathway and autophagy. |
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