Dissertation > Medicine, health > Obstetrics and Gynaecology > Gynecological surgery

Study the Effect of Transient Intervention of HMG on the Result of Ovary Transplantation in Mice

Author SunJing
Tutor WangYanRong
School Ningxia Medical University
Course Human Anatomy and Embryology
Keywords Ovary Transplantation Gn vascularization Evaluation of Viability
CLC R713
Type Master's thesis
Year 2009
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The cryopreservation and transplantation of ovarian tissue, as a way of conservation and recovery the fertility of the female, give hope to those who suffer from endocrine dyscrasia and fertility deprivation as a result of POF or ovriectomy or congenital ovarian dysgenesis. The method of ovarian cortex transplanted after cryopreserved without vascular anastomosis is easier, more economic than that of ovary organs, and the former is easier to be performed and can avoid the forming of thrombus which much often occurs after the vascular anastomosis. However, the ischemic hypoxia and ischemic–reperfu sion injury after the operation often cause great loss of primordial follicles, which results in low survival rate and worse function and shorter life span of the transplanted ovary. Some researches believe that the secretion of Gn after the ovary transplanted is helpful to the revascularization of the graft. Purpose This research is to explor not only the influence of the revascu- larization caused by the intervention of HMG during the time of in vitro and transplanting but also some mechanisms.Methods①Observe the VEGF expression in ovarian tissue by immunohistochemical method after cultured the hemi-ovaries of juvenescent mice in the culturing medium that contain HMG 0.00IU/mL, 0.15IU/mL, 0.30IU/mL and 0.60IU/mL for 12h, 24h and 48h in juvenescent mice;②The hemi-ovaries are cultured for 3h in vitro before heterotrophic autotransplantation, then to observe the differences of blood vessel density in different transplanting sites,and bloodstream feedthrough,and the tissue fluid exudation quantity around and inside of the graft after transplanted for 24h、36h by way of perfusion with prepared Chinese ink; the positive rate of apoptosis protein was evaluated by the expression of Bcl-2, Bax and Caspase-3 in each group after transplanted for 72h;observe the effect of different concentration of HMG on the follicles surviving after transplanted for 1 month.Result 1.There exist differences in revascularization after the intervention of HMG in different transplanting sites: The density of blood vessels of Kidney(406.90±61.53) is more greater than the neck subcutaneous tissue(133.38±2.26), (P<0.001); It can be seen that the blood vessels throughout the kidney to the graft after transplanted for 24h, but not in the neck subcutaneous tissue group after transplanted for 36h.There were more normal morphologically normal follicles (9.38±5.82) can be observed than that of the neck subcutaneous tissue group(5.57±1.12), (P<0.05).2.There is significant difference in VEGF expression of the ovarian tissue after cultured in vitro and bloodstream feedthrough after transplantated with different HMG concentration.(1)The quantity of VEGF expression, which is higher than that of normal control group, can be observed at each time phase; the quantity in 24h and 48h is significant higher than that of HMG0.00-CG.The higher the HMG concentration grows the higher quantity of VEGF expression in HMG intervention group in each period incease. The peak can be found after cultured for 12h in the non-intervention group (0.00IU/mL HMG),low(0.15IU/mL HMG) and the middle concentration of HMG group(0.30IU/mL HMG); with longer culturing time, there shows mini- decrease in the quantity. However, the highest quantity of VEGF expression can be found during each time in the high concentration of HMG group (0.60IU/mL HMG), and the peak appears after cultured for 24h (95.52± 12.05); There is great significant difference in the quantity compared with that of other groups, (P<0.001).The effect of different HMG concentration on grafts:①The higher the HMG concentration increase the exudation area after transplantation grows,and significant differences can be found in the high(0.08±0.04),middle(0.06±0.03)and low concentration group (0.03±0.02) (P<0.001);The blood supply can be seen between the grafts and recipients by way of ink-perfusion and immunohistochemical after transplanted for 24h in the middle and high concentration of HMG groups. The blood vessels between kidney tissue and grafts were seen to be filled with ink in HMG intervention groups after transplanted for 36h.The developing follicles at different stages are preserved perfectly could be seen in the survival ovarian grafts in each group by histological examination. The effects of transient intervention of HMG on the development of follicles in the grafts:①The ratio of normal follicles and positive expression of follicle apoptosis with transient and high concentration intervention of HMG after transplanted for 72h is as follows: The number of normal follicles in CTG(5.00±1.10) is the least and showed significant difference compared with either FTG(9.57±2.82) or HMG0.60-CT G(9.00±1.98),P<0.001.While no significant difference was indicated between FTG and HMG0.60-CTG,P>0.05. It is the lowest of the positive expression of Bcl-2 in CTG(48.08±19.84).There was significant difference compared with other groups, P<0.001.While highest of Bax were found in CTG (34.61±17.00),and significant differences can be found compared with FTG, P<0.05. Positive expression of Caspase-3 in CTG(54.41±44.10) is the highest in which significant differences can be found compared with other groups, P<0.05. No significant differences can be found obout the positive expression of Bcl-2, Bax and Caspase-3 between HMG0.60-CTG and FTG.②The recovery of estrous cycle in CTG(6.83±0.75) is longer after being autotransplanted,and significant differences can be found compared with other groups, P<0.05; However there was no significant differences in the recovery rate of estrous cycle between other groups. The total number of survival follicles in HMG intervention group is evidently higher than that in non-intervention group after transplanted for 1 month,and the number of survival follicles in HMG0.15-CTG was higher than that in other HMG intervention groups and FTG, P<0.05; While the total number of survival follicles in HMG0.60-CTG (6.91±2.51)was significant lower than that in HMG0.15-CTG(9.36±3.02) and FTG(8.04±2.85),P<0.05.Conclusion①The density of blood vessels in the position of transplanting will influence the effects of revascularization after the transplantation, which was caused by Gn;②It can shorten the time of revascularization between the graft and recepient with HMG transient and extrasomatic intervention before transplantation and immediately conducted on the sites of transplanti- on during the operation.③The expression of VEGF becomes higher as the concentration of HMG increasing; the same as the quantity of exoserosis plasm exhidrosis in the site of transplanting.④The intervention of HMG prior to transplanting is helpful to follicles survival in graft, and HMG0.15-CTG is better than HMG0.30-CTG and HMG0.60-CTG in keeping the number of follicles.

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