The Effects Of Bone Marrow Mesenchymal Stem Cells’ Transplantation On Mice’s Spermatogenic Dysfunction

Background and Objective：Infertility is a disease which affects fifteen percent couples all over the world.Among them, male factors as the main reasons account for thirty percent, while bothof forth percent couples have problems. The pathophysiology of male infertility isstill uncertain, so for the patients whose pathogen are certain, they are givensymptomatic treatments，while for the patients whose causes are uncertain, they aregiven experiential treatments，whose effect are poor most time. During recent years,assisted reproductive technology (ART) developed rapidly, but ART has somedisadvantages, including invasive, high economic burden and potential healthy risk.In addition, for those couples one of who can not produce functional gamete, there arenot any treatment but turning to donors.With the development of stem cells’ research in male infertility in recent years，it was reported that stem cells could produce male germ cells in vitro and vivo, whichprovide new aspects for male infertility’ treatment and pathophysiological research.Our research’s aim is to investigate whether bone marrow mesenchymal stem cells’transplantation can treat male mice’s spermatogenesis dusfunction and it’s possiblemechanism in order to offer more treatment option for clinical work.Methods：Seventy-five healthy Wistar mice were divided into five groups equally. Everygroup has fifteen mice. The group A is normal control group; the group B is controlgroup of low dose irradiation; the group C is control group of high dose irradiation;the group D is BMSCs treatment group of low dose irradiation; the group E is BMSCs treatment group of high dose irradiation. Both B and D groups were given60Co γ irradiation for one time, while C and E groups were given60Co-γ irradiationfor twice every one week. After the models established sucessfully, the mice of the Dand E groups were given BMSCs’ transplantation, and then they were raised for eightweeks. Eight weeks’ later, they were killed, and we got their whole weight and theweight of testis, and then calculated testis index. We determined the level of serumfollicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T)by enzyme-linked immunosorbent assay (ELISA). We evaluated pathology of testisby Haematoxylin and eosin stain (HE). Then we used reversetranscription-polymerase chain reaction(RT-PCR) to evaluate the level of GFP、Inhibin B、P450scc and VASA. Finally, we use double fluorescence immunostain toevaluate the level of GFP and VASA.Results：After irradiation, both the weight and testis index of B and C group weredecreasing, which had statistical significance（P＜0.05）,while for the D and E group,there was not difference for the weight, but for testis index, it was still decreasing,which had statistical significance（P＜0.05）. As for sex hormone, compared with Agroup, the level of FSH and LH of the other groups were increasing, and for T, it wasdecreasing,both of which had statistical significance（P＜0.05）.After treatment, thelevel of LH were decreasing, while T increasing, both of which had statisticalsignificance（P＜0.05）. But for FSH, C group had statistical significance（P＜0.05）,while E group did not have(P＞0.05). As for HE, after irradiation, seminiferoustubules were destroyed greatly, where there were rare sertoli cells and spermatogonia,and C group was worse. After treatment, the spermatogenesis function of D groupwas recovered partly, while E group not. As for RT-PCR, after irradiation, the levelsof Inhibin B、P450scc and VASA mRNA were very low,which we can not detect in Band C group.Then after treatment, the level of Inhibin B、P450scc and VASA mRNAin D group were increaing, while for E group, only the level of P450scc mRNA wasincreasing, and the level of Inhibin B and VASA mRNA did not improve.In final, the results of double fluorescence immunostain were similar to RT-PCR.Conclusion：BMSCs’ transplantation can improve mice’s spermatogenic function for thosemice which were damaged lightly, but for those mice which were damaged worse, itwas not improved owing to the sertoli injury.