| BACKGROUND: Aging is accompanied by a variety of chronic diseases,is the inevitable process of human life.Now,the number of aging population is increasing year by year,people's desire to delay aging and prolong the health life is more and more intense.Despite the rapid development of science and technology,it is still not able to fully explain the internal mechanism of aging.Metabonomics rely on advanced analysis technology platform,to obtain the information of endogenous small molecule metabolites,then analysis of a series of biological events have been explained in the body from the aspect of the metabolites.Currently the study on metabonomics has been gradually applied to various complex diseases,it is also an opportunity for aging research.Traditional Chinese medicine Scutellaria baicalensis georgi,has a long history and abundant resources.Modern pharmacological research also showed that Scutellaria baicalensis georgi has the effect of antioxidant,anti-inflammatory,antitumor,free radical scavenging and nerve protection and so on.Previous study showed that Scutellaria baicalensis extract could improve the activity of antioxidant enzymes so as to achieve anti-aging effect.In the early stage of our research group,D-galactose induced aging model of rats was used to evaluate the anti-aging activity of Scutellaria baicalensis georgi.On the basis of these studies,now,our research group introduced the model organism Drosophila melanogaster.Drosophila has short lifespan,easy feeding,low cost,simple experiment operation and other advantages.Currently,it is the aging research commonly used model organism,however,it is rare to study aging and gender gap in aging process of Drosophila melanogaster based on metabonomics and the studies on the evaluation of the anti-aging activity of Scutellaria baicalensis Georgi by Drosophila melanogaster have not been reported.OBJECTIVE: In order to further study of aging and gender gap in aging process,our research investigate the the metabolism variation between male and female Drosophila melanogaster in aging process.In order to evaluate the anti-aging activity of Scutellaria baicalensis georgi,our research carried out the metabonomics study about alcohol extractfrom Scutellaria baicalensis georgi prolonging lifespan of Drosophila melanogaster based on 1H-NMR.METHODS: 1.The same batch of Drosophila samples were fed under the same conditions,at four time points(days 3,15,30,and 45),we collected Drosophila samples for 1H-NMR metabonomics analysis.Dynamic monitoring the change trend of Drosophila melanogaster metabolic profiles and endogenous metabolites.First of all,the metabolic profile changes of Drosophila melanogaster during 3d to 45 d were carried out combined with principal component analysis(Principal component,analysis,PCA)and partial least squares discriminant analysis(Partial least squares-discrimination analysis,PLS-DA).Secondly,statistical analysis of the Drosophila metabolites changes with age and calculate the correlation coefficient of metabolites with age,in this way the endogenous metabolites change associated with aging process of male and female Drosophila melanogaster were screened out,and pick out the common parts of male and female flies to be senescence associated biomarkers.Finally,the metabolic pathway of these metabolites were enriched to find the metabolic pathway related to Drosophila aging process.2.3d?15d?30d and 45 d the metabolic profile differences between male and female Drosophila melanogaster were analyzed by metabonomics.Firstly,the differences metabolites between male and female flies in each age group were found by multivariate statistical analysis.Then find out the metabolites(in 3d,15 d,30d and 45 d four age states)which were found lower or higher in male(female)consistently and significantly were the most relevant metabolites to the gender differences,were considered to be gender gap related metabolites,and the common part of the biomarkers associated with aging process were considered as metabolites associated with gender gap in aging process.Finally,based on these metabolites to find the metabolic pathways related to the gender gap in aging process.3.On the basis of a preliminary understanding of the metabolic changes during the aging process of Drosophila melanogaster,the anti-aging activity of Scutellaria baicalensis Georgi was evaluated by the life span study and metabonomics study.First of all,the Drosophila melanogaster culture medium with different concentrations(0.1 g/L,1g/L,5 g/L)was used to study the effects of Scutellaria baicalensis Georgi extract on the average life span and life span of Drosophila melanogaster.Then find out the regulation metabolites of Scutellaria baicalensis Georgi in Drosophila melanogaster by metabonomics and multivariate statistical analysis.Finally,corresponding metabolic pathways of Drosophila melanogaster were enriched and its biological significance was explained.RESULTS: 1.The results of metabolic profiling showed that the metabolic profiles change trend in aging process were different between male and female Drosophila melanogaster.The analysis of PCA and PLS-DA showed that the metabolic profile of female flies 3d,15 d,30d and 45 d were separated from each other,while in male flies,the metabolic profile of 30 d and 45 d are close together,3d?15d and 30 d separated from each other.Clearly we can see,male Drosophila metabolic profile from 3d to 15 d to 30 d changed significantly,but the change from 30 d to 45 d is not obviously,this may be 30 d in male Drosophila melanogaster has entered the aging stage.The changes of metabolite analysis showed that there are some similarities and differences in the metabolites changes from 3d to 45 d between male and female flies as follows:(1)between the male and female flies,part metabolites(such as: methionine,glucose etc.)change trend with age entirely the same and only the levels of metabolites have differences in male and female(the same point between male and female);(2)in female flies,some metabolites(such as isoleucine,valine,alanine,tyrosine)from 3d to15 d,there is a rise or flat stage in content,but in male flies it is not the existence of such phenomenon(different point between male and female)(3)almost all of the metabolites content is different between male and female flies at the same life stage,and most of them showed significantly lower in male flies(how would male and female flies be different).Aging related metabolic markers of Drosophila including: isoleucine,valine,alanine,glutamine,succinate,methionine,glycine,cysteine,tyrosine,tryptophan,phenylalanine,histidine,xanthine,DMA.Metabolic pathways are mainly related to the amino acid synthesis and metabolism,purine metabolism,tricarboxylic acid cycle.In addition,changes of some metabolites also suggest that the aging process is accompanied by mitochondrial dysfunction.2.Studies on the metabolism of male and female Drosophila melanogaster showed that there were significant differences in the metabolic profiles between male and female flies during 3d?15d?30d and 45 d these four life stage.Find out the metabolites associated with gender gap in aging process including: isoleucine,valine,methionine,phenylalanine,cysteine,succinate,DMA and metabolic pathways are mainly involved with the biosynthesis and metabolism of amino acids and tricarboxylic acid cycle.In addition,the difference in the content of some metabolites between male and female Drosophila melanogaster also imply the gender differences in the mitochondrial function of Drosophila melanogaster.3.Lifespan experiments showed that Scutellaria extracts 1g/L and 5g/L administration group can significantly prolong the average life expectancy and the highest life expectancy of female flies,the 5g/L group can significantly prolong the average life expectancy and the highest life expectancy of male flies.Metabonomics study results showed that Scutellaria baicalensis Georgi can adjust the content of isoleucine,valine,acetate,glutamine,phenylalanine,cysteine,glycine,succinate,lactate and glucose of Drosophila,and may improve the damage of mitochondrial function in this way to extend the life of the fruit flies.CONCLUSION: 1.NMR based Metabonomics study on fruit fly showed that the change trend of metabolic profiles and metabolites of male and female Drosophila melanogaster were different in aging process.Male flies may enter senescence at 30 d,and female flies may be later.In this study,14 markers of aging related metabolic markers and aging related metabolic pathways were screened out by the combination of the study of male and female flies.2.7 metabolic markers and related metabolic pathway about gender gap in aging process were found by using metabonomics research method and multivariate statistics.3.Both the average life expectancy and the highest life of male and female flies can be significantly extended by adding the Scutellaria Baicalensis extract,in addition,themetabolites and metabolic pathways which can be adjusted by Scutellaria Baicalensis Extract were be found.4.Drosophila melanogaster aging process metabonomics study showed that the aging process of Drosophila accompany with mitochondrial dysfunction;gender gap in aging process metabonomics study showed that the mitochondrial function may exist gender differences;metabonomics study of Scutellaria baicalensis Georgi anti-aging activity showed that Scutellaria baicalensis Georgi may be able to repair the damage of mitochondrial function. |
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