Preliminary Study On Molecular Mechanism Of Quercetin On Anti-aging In Simocephalus Vetulus

Enzyme activity detection,proteomics technology,real-time quantitative PCR,gene cloning and bioinformatics analysis were used in this study to detect the anti-aging effects of quercetin on Simocephalus vetulus.The detailed results were summarized as follows: 1 Quercetin can prolong the lifespan of Simocephalus vetulus,keep it healthy and have no negative effect on growth-related parameters.Newborn Simocephalus vetulus was exposed to quercetin(0,1,2.5,5,10 mg/L),and the quercetin solution was changed every 24 hours.The number of deaths and molting,reproductive quantity and reproduction time were recorded until all individuals died.Survival curve was drew and the growth-related parameters such as net reproductive rate,intrinsic growth rate and generation cycle were used formulated to calculate,the body length was measured under the microscope,the metabolic indexes such as ACP,AKP,T-CHO,NEFA,TG,Glu,AMS and CMCA were detected with enzyme activity kit.The results showed that appropriate concentration of quercetin can prolong the absolute lifespan and average lifespan of Simocephalus vetulus,and has no negative effect on the net reproductive rate,intrinsic growth rate,generation cycle and the body length of Simocephalus vetulus.Quercetin can regulate the digestive enzymes activity,keep a healthy status of lipid metabolism and immunity in Simocephalus vetulus.These results reveal that quercetin can prolong the lifespan of Simocephalus vetulus,keep it healthy and have no negative effect on growth-related parameters.2 Quercetin by affecting carbohydrate metabolism of Simocephalus vetulus for its anti-aging effect.Newborn Simocephalus vetulus was exposed to quercetin(0,1 mg/L)for 7 days.The protein sequencing alignment of Simocephalus vetulus was used by proteomics technology,the credible proteins and differential proteins were selected by bioinformatics technology,and bioinformatics technology also used to Gene Ontology enrichment analysis and KEGG Pathway enrichiment analysis.The results showed that 156 differential proteins were selected after quercetin treatment,and there were 84 significantly up-regulated proteins and 72 significantly down-regulated proteins.Quercetin significantly enhanced the key enzyme activities of carbohydrate metabolism led by starch and sucrose metabolism in Simocephalus vetulus.These results suggest that quercetin by affecting carbohydrate metabolism of Simocephalus vetulus for its anti-aging effect.3 Quercetin can not only decrease the degree of oxidative damage in Simocephalus vetulus but also increase the expression of anti-oxidant enzyme genes and heat shock protein genes.Newborn Simocephalus vetulus was exposed to quercetin(0,1,2.5,5,10 mg/L)respectively for 7 days and 14 days,exposed to quercetin(0,1 mg/L)for 21 days.The degree of protein carbonylation and the content of MDA were detected by kit.The genes expression of Simocephalus vetulus were analyzed by real-time quantitative PCR.The results showed that the level of protein carbonylation and MDA in Simocephalus vetulus was significantly decreased after quercetin treatment.The expression of Mn-SOD,Cu/Zn-SOD,GPX,GST and HSP70,HSP90 were significantly increased after quercetin treatment for 7 days.The expression of CAT,HSP70 and HSP90 were significantly increased but the expression of GPX and GST were significantly decreased after quercetin treatment for 14 days.The expression of HSP90 was significantly decreased and the other genes were not significantly changed after quercetin treatment for 21 days.These results imply that quercetin can not only decrease the degree of oxidative damage in Simocephalus vetulus but also increase the expression of anti-oxidant enzyme genes and heat shock protein genes.4 The gene full length of Mn-SOD and Sir2 in Simocephalus vetulus is successfully cloned and the expression of these two genes both significantly increase after quercetin treatment.Newborn Simocephalus vetulus was exposed to quercetin(0,1,2.5,5,10 mg/L)respectively for 7 days and 14 days,and exposed to quercetin(0,1 mg/L)for 21 days.The full length of target genes was cloned by gene cloning technology,bioinformatics technology was used to analyze the c DNA sequence and predict protein structure of target genes,the expression of target genes were analyzed by real-time quantitative PCR.The results showed that the full length of Mn-SOD was 736 bp,including 56 bp 5'UTR,32 bp 3' UTR,648 bp ORF and encoding 215 amino acids.The full length of Sir2 was 1974 bp,including 19 bp 5'UTR,140 bp 3' UTR,1815 bp ORF and encoding 604 amino acids.The protein encoded by the Mn-SOD gene is a homotetramer and Mr=24.36 k Da,p I=8.42.The protein encoded by the Sir2 gene is a heterodimer and Mr=67.38 k Da,p I=4.69.The expression of Mn-SOD gene and Sir2 gene were both significantly increased after quercetin treatment.The results indicate that we successfully clone the gene full length of Mn-SOD and Sir2 in Simocephalus vetulus and the expression of these two genes both significantly increase after quercetin treatment.