Study On The Mechanism Of Phycocyanin In Ovarian Function Rescue In Naturally Aged Mice By Using Rna-seq

For adult women,their fertility declines with age.Ovarian aging,including the decrease in the number of follicles and the functional decline,have become important causes of fertility decline.Rescuing the aging ovary is an important strategy to increase female fertility.C-Phycocyanin(PC)is a kind of protein of Spirulina platensis with antioxidant,anti-inflammatory and immune-enhancing properties,which has been widely used in medical and biological fields.Our previous study used D-galactose(D-gal)aging mice model,indicating that PC could improve ovarian function of aging mice by inhibiting the Reactive Oxygen Species(ROS)pathway;Furthermore,RNA-Sequencing(RNA-Seq)technology was used to screen enrichment pathways that resulted in differentially expressed genes in the attenuated and attenuated improved groups to increase fecundity.But in a natural aging mouse model,whether PC can improve female fertility and the effect and mechanism of PC have not been known.In this study,6-weeks-old ICR mice are taken as the young group,the 8.5-monthold mice as the natural aging group and 8.5-month-old mice treated by gastric PC as treated group.After 45 days of the treatment,RNA-Seq technique is used to analyze the changes of ovarian expression at transcriptional level.The following conclusions are drawn:(1)The data of ovarian transcriptome of Young,Aged and Aged+PC three groups are obtained through high-throughput generation sequencing.After filtering data,the total amount of 58.14 G data is obtained,with an average of 92.85% compared to the genome.9 samples had Q30 index above 93.20%,indicating that the quality of sequencing is high and the follow-up analysis results are reliable.(2)The ovarian expression profiles of natural aging mice are constructed,and 597 different genes are identified,in which the number of up-regulated genes is 252 and the number of down regulated genes is 345.According to GO analysis,differentially expressed genes are mainly enriched in circulating immune globulin mediated humoral immune response,complement activation classical pathway,B cellmediated immunity,etc.In the cell components enriched mainly in blood particles,immunoglobulin complex cycle,chromosome condensation movement;in molecular function mainly enriched in antigen binding,immunoglobulin receptor binding and so on.According to the KEGG analysis of differentially expressed genes in cell cycle,cell adhesion molecules are significantly enriched.(3)Compared with Aged group,13 common differentially expressed genes are identified between Young group and Aged+PC group.According to the GO analysis,the common differentially expressed genes are mainly enriched in the biological processes of the cell response to interferon-?,cytokine response,innate immune response,defense response to protozoa and so on;The cellular components are mainly enriched in vacuolar membrane symbiont,host cytoplasm and part of host cell.(4)Based on the differentially expressed genes between the Young group and the Aged group,and combined with the RT-qPCR results,Aurka,Gdf9,Mests and Rad5 gene is up-regulated,among which Gdf9 and Rad51 are significantly up-regulated,and Cd74,Cxcl10,Nlrc5 and Zbp1 genes are down-regulated significantly.This is consistent with the results of RNA-Seq.The up-regulated genes are involved in oocyte microtubule formation,follicle development,growth-promoting and DNA repair,and down-regulation genes are involved in inflammatory response,hypomethylation involved in the regulation of aging and so on.We speculated that PC could improve ovarian function,delay aging and provide a theoretical basis for PC to resist ovarian aging and increase fertility.