Effects Of Gonadotropins On The Expression Of VEGF And Survivin In The Mouse Ovaries And The Birth Rate After Vitrification Cryopreserved Transplantation

It’s becoming increasingly popular for young and adolescent females to preservetheir fertility through transplantation after the ovaries being kept under profoundhypothermia. However, the great loss of ovarian follicles after avascularity compatiblegrafting shortens the functional life of the transplanted ovaries. Therefore, to findeffective ways to solve the problems of the loss of ovarian follicles after transplantationhas become a hotspot of research for females to preserve fertility in vitro. The studygroup preliminary work shows the brief intervention of HMG in vitro enable the micetransplanted ovarian perfusion time in advance of FSH intervention of sheep, frozentransplanted ovarian follicles showed improved survival, these are just from theperspective of follicle survival observed Gn role, yetexperimental evidence for the lackof save observation mechanisms and fertility. LH and FSH Intervention mouse ovaries invitro, observation of angiogenesis and anti-apoptosis related gene expression andintervention birth after orthotopic transplantation of ovarian vitrification, hoping topromote gonadalhormonal intervention to improve ovarian transplantation after fertilityprovide a scientific basis.ABSTRACTPartⅠ Effects of LH and FSH Intervention on the Expressions of CulturedMouse Ovarian VEGF and SurvivinObjective Explore the effects of the intervention of LH and FSH on theexpressions of ovarian angiogenic factors of VEGF and Survivin cultured in vitro. Methods Observe the expressions and correlation analysis of VEGF and Survivinby immunohistochemical method,RT-PCR,Elisa after the hemi-ovaries of juvenescentmice had been cultured in the culturing mediums that contain LH,FSH of0.00IU/ml and0.15IU/mL,0.3IU/ml,0.6IU/ml for3,6,12,and24hours.Result The expressions of VEGF in FSH and LH group by immunohistochemicalmethod and Elisa are higher than CG group at6h,12h. The expressions of Survivin inFSH and LH group by immunohistochemical method,RT-PCR,Elisa are higher than CGgroup at12h. The expressions of VEGF in LH-0.3is much higher than others group at24h, P <0.05. The expressions of VEGF and Survivin in FSH and LH group aresignificant correlation, Pearson correlation is0.792, P <0.001.Conclusion LH intervention in vitro can increase expressions of VEGF andSurvivin and correlation analysis are significant. The expressions of VEGF and Survivinin0.3IU/mL LH is significantly better than any others group.PartⅡ Effects of LH Intervention on the Birth Rate of vitrifiedCryopreservation Mice after Heterogeneous Orthotopic Transplantation of TheirOvariesObjective To study the effects of LH intervention on the survival and functionalrecovery of the mouse ovaries after heterogeneous orthotopic transplantation.Method The C57BL/6J mice (black hair) are ovary donors, the C57BL/6J-Tyrc-2J-J white hair mice (mutants) are ovary receptors and they are randomly divided intothe control group, the fresh transplantation group, the cryopreservation transplantationgroup without LH intervention and the cryopreservation transplantation group with LHintervention with6white hair mice and6black hair mice in each group to study theirbirth. For the control group, the white hair male mice and the black hair female mice areput into the same cage respectively. For the fresh transplantation group, the complete bilateral ovaries of the black hair mice are dissected from their ovarian bursa andtransplanted into the bilateral ovarian bursa of the white hair mice. For thecryopreservation transplantation group without LH intervention, the ovaries of the blackhair mice are dissected in the same manner and transplanted three days after vitrifiedcryopreservation. For the cryopreservation transplantation group with LH intervention,the dissected ovaries are transplanted three days after vitrified cryopreservation in thevitrification solution of0.3IU/ml LH. The protection efficient of the birth rate isobserved through the estrous cycle and the births of the black hair mice aftertransportation.Results The estrous cycles of the four groups are all recovered. For the freshtransplantation group, the recovery time is6.5±1.04days, for the cryopreservationtransplantation group,9.33±1.21days and for the cryopreservation transplantation groupwith LH intervention7.16±1.17days. It takes longer for the cryopreservationtransplantation group to recover the estrous cycle than the fresh transplantation groupand the cryopreservation transplantation group with LH intervention and the difference issignificant (P <0.05). All the mice put in the same cage get pregnant normally. Theaverage bearing capacity is5.83±1.94mice per brood in the control group,4.67±1.37inthe fresh transplantation group and2.83±0.98in the cryopreservation transplantationgroup with significant difference (P <0.05). The average bearing capacity is4.16±1.72mice per brood in the cryopreservation transplantation group with LH intervention thesame as that in the fresh transportation group with no significant difference.Conclusion The vitrification solution with0.3IU/ml LH intervention is helpful tothe recovery or the estrous cycle and birth rate of the isoplastic mice after heterogeneousorthotopic transplantation of their ovaries.