Evaluation Of Anti-aging Effects And Mechanism Research Of Trifoside Based On Metabonomics

Aging is an inevitable stage of biological process,accompanied by a variety of aging and degenerative diseases,such as cardiovascular and cerebrovascular diseases,diabetes,respiratory diseases,cancer and so on.It is of great significance to delay aging and improve the quality of life.Asperuloside is an iridoid isolated from Rubiaceae,Eucommiaceae,Galium and Galium plants.Recent pharmacological studies have shown that asperuloside has many pharmacological activities,such as antiinflammatory,analgesic,antihypertensive and anti-cell chromosome mutagenesis.There are few studies on the anti-aging effect of asperuloside.This paper studied the antioxidant and anti-aging effects of asperuloside in vivo were evaluated in fruit flies.The aim of this study was to investigate the antioxidant and anti-aging effects and mechanisms of asperuloside,and to provide theoretical basis for the research and development and utilization of asperuloside.The main results of the experimental study are as follows: 1.Prolongation of life span of asperuloside on wild drosophila melanogaster and its mechanismWild drosophila melanogasters were used as a model animal to investigate the effect of asperuloside on the mean life span,maximum life span and half death time of male drosophila melanogaster in natural aging model.At the 15 th and 35 th days of age,drosophila melanogaster was determined the CAT activity.Acute oxidative injury experiment of hydrogen peroxide was carried out at 35 th days of age to study the protective effect of citrinin on oxidative stress.At the ages of 0,15 th and 35 th days,quantitative real-time PCR was used to determine the mRNA expression of related genes in drosophila melanogaster.The results showed that the mean lifespan,maximum lifespan and half of death time of drosophila melanogaster were prolonged in drug groups.CAT activity in male drosophila melanogaster treated with different doses(0.15 mg/mL,0.30 mg/mL)was higher than that in control group,and showed a dose-effect relationship.Compared with the control group,0.30 mg/mL citrin could significantly prolong the lifespan of drosophila melanogaster and reduce the acute oxidative injury in vivo.The mRNA expressions of SOD2,CAT,SOD1,MTH and Rpn11 genes in drosophila melanogaster were significantly up-regulated in different doses of asperuloside at 35 th days of age.2.Prolongation of life span of asperuloside on w1118 fuit flies and its mechanisimThe w1118 fruit flies were used as the model to investigate the effect of different diet on the longevity of female and male fruit flies in the restriction model,and three different concentrations of food were selected for subsequent experiments.The effects of asperuloside on the lifespan of male and female fruit flies were studied under sucrose/yeast diet with different concentrations(0.1 SY,1 SY,2 SY).To investigate the effects of asperuloside on taste behavior of w1118 fruit flies at 6th days of age,the effects of asperuloside on the lifespan of fuit flies w1118 at 32 th days of age of hydrogen peroxide model,and the effects of asperuloside on SOD1 activity in fruit flies at 32 th day of age.Sucrose/yeast feeding at different concentrations(0.1 SY,0.25 SY,1 SY,2 SY)limits the lifespan of fruit flies.The results showed that the lifespan of male and female fruit flies could be effectively prolonged by controlling sucrose/yeast concentration within a certain range.Compared with the control group,both the 0.30 mg/mL asperuloside group and the rapamycin group extended the mean lifespan,maximum lifespan and half of the death time of both male and female fruit flies.The 0.30 mg/mL asperuloside group increased the antioxidant stress capacity of both male and female fruit flies,but had no significant effect on the SOD1 activity in fruit flies.The changes in the lifespan of w1118 fruit flies caused by these asperuloside were unlikely to be related to the caloric restriction.The taste behavior measurement of fruit flies showed that compared with the control group,there was no difference in the degree of abdominal redness and food intake in the 0.30 mg /mL glycosides group.3.Analysis of w1118 fruit fly metabolites based on GC-MSFirstly,a quantitative and qualitative analysis method for metabolites of w1118 fruit flies was established by GC-MS.The results showed that the GC-MS method for the determination of w1118 fruit fly metabolites weas feasible.The contour model of metabolites of fruit flies was established by PCA and PLS-DA.The displacement test chart to test PLS-DA model.VIP was calculated to find biomarkers related to aging and quantitatively describe the importance of biomarkers on metabolic changes in fruit flies.There were significant differences between the control group and asperuloside group.The PLS-DA model of fruit flies fed with different sucrose/yeast had good separation degree.The changes of frui fly metabolites were quantified by VIP value.The biomarkers related to the anti-aging effect of asperuloside were pyruvic acid,L-valine,L-(+)-lactic acid,alanine,β-alanine,succinic acid,D-malic acid,L-cysteine,α-pentanedic acid,glycerol 1-phosphate,glycerol,glycerol and glycerol 3-phosphate,citric acid,myristic acid,stearic acid,D-galactose,inosine and ergosterol.