Study On The Mechanisms Of Human Seminal Exosome And The Pentacle Gold-copper Alloy Nanocrystals Delivery Carrier In Asthenozoospermia

About 15%to 20%couples faced infertility problems all over the world,approximately 15%of cases male infertility is caused by asthenozoospermia.The clinical manifestations of asthenozoospermia are poor sperm motility,asthenozoospermia usually implies little or nothing about the actual pathogenesis of the problem,which is difficult for diagnosis and treatment,and the fertilization rate after ICSI with severe asthenozoospermia sperm is also very low,Therefore,it is significance to study the pathogenesis of asthenozoospermia.there are a large number of exosomes in human seminal plasma,the mechanisms of seminal exosome regulation the sperm motility also remains to be further elucidated.In this study,we have identified 3699 proteins in seminal exosome via using iTRAQ proteomics techniques by seperating the exosomes from normal and asthenozoospermic semen.Compared with normal group,11 proteins were significantly upregulated(>1.2)and 80 proteins were significantly downregulated(<0.833)in asthenozoospermia seminal exosome.The proteomic result was validated by Western blot,functional bioinformatics analysis show these differential proteins were mainly in transmembrane transport and metabolic.we found TRPV6 in epididymis caput start transfering to spermatozoa by epididysome,and the TRPV6 decreased significantly in asthenozoospermic seminal exosome.these results indicated that seminal exosome secretory disorders may be one of the cause of asthenozoospermia.Gold nanoparticle has been attracted considerable attention and application in the field of biomedicine,many kinds of gold nanoparticle have been widely used in hyperthermia,drug-targeted transport,biological image,cell labeling and gene transport,There are multiple barriers in vivo,such as blood-testis barrier,blood-placenta barrier,and blood-brain barrier,However,overcoming biological barriers is a hard and inevitable problem,which restricts medical applications of nanomaterials in vivo.Seeking for an efficient transportation to penetrate biological barriers is a common need.Here,we pay close attention to the blood-testis barrier.We synthesize pentacle gold-copper alloy nanocrystals,and found that the pentacle gold-copper alloy nanocrystals not only could enter GC-2 cells in vitro in a short time,but also could overcome the blood-testis barrier and enter male germ cells in vivo.We treated the GC-2 cells and injected the ICR male mices by tail vein with gold-copper alloy nanocrystals,found the gold-copper alloy nanocrystals did not affect the viabilities and apoptosis of GC-2 cells,and there were no significant differences in testicular morphology between pentacle-treated mice and the control group,indicating that the gold-copper alloy nanocrystals have good biocompatility.Furthermore,we demonstrated that the entrance efficiency would become much higher in the development stages or 43? water bath after 10 min treatment.The results also suggested that the pentacle gold-copper alloy nanocrystals could easier enter to germ cells in the pathological condition.This system could be a new method for theranostics in the reproductive system.and which provides a new direction for male infertility drug transport.