| Objective: Premature ovarian failure is the leading cause of infertility in women today.Because the original follicle reserves are small and the maturation disorder results in a very low natural conception rate,how to activate the remaining original follicles becomes the key to treatment.In 2013,Japanese scholars obtained a pregnancy through the technique of activating fragmented ovarian cortical in vitro activator co-culture to activate dormant follicles.In 2016,Chinese scholars also achieved successful pregnancy only through fragmented cortical transplantation.Because the safety of in vitro activators is unknown,and there is no rapid vascular anastomosis and blood supply establishment after ovarian cortical transplantation,it is in an early state of ischemia and hypoxia,which can cause related issues such as follicle loss.In this experiment,the ovaries of premature aging mice were stimulated in vivo,the number of follicles in each group before and after treatment,and the hormonal changes before and after treatment were explored to explore a new method of assisted reproduction and its feasibility in patients with premature ovarian failure.Materials and Methods: In this study,cyclophosphamide was used to construct a mouse model of premature aging.All mice were divided into 6 groups: blank control group,model control group,experimental group A,experimental group B,elderly control group,and elderly experimental group.After taking the posterior orbital venous blood for backup,the experimental group acupunctured the bilateral ovaries25 times each with a depth of about 1 mm.The control group only needed to explore the ovaries to close the abdomen.Except that the experimental group B was ovulated on the 10 th day after operation,and was sacrificed on the 13 th day,the other groups were given ovulation on the 5th day after operation and sacrificed on the 8th day.The bilateral ovarian tissues and venous blood were collected,the changes of hormone levels before and after treatment,and the HE staining of ova rian tissues of mice in each group were observed to observe the number of follicles at various levels.The remaining model mice were taken,and the ovarian tissues weretaken for immunohistochemistry to detect the expression of YAP protein 1 hour after intervention.Research result: 1.The weight of mice after modeling was 26.83±0.68 g,which was statistically significant compared with 28.5±0.57 g before modeling(P <0.05).2.The estrous cycle of the modeled mice was 12.4±1.6d,and the difference was statistically significant compared with the blank control group(P <0.05).3.The blood FSH concentrations in the experimental group A and experimental group B after intervention were 12.15±3.20mIU/mL and 12.36±0.91mIU/mL,which were lower than those before the intervention,and the difference was statistically significant(P <0.05).The blood FSH in the elderly experimental group after the intervention It was 11.34±3.58mIU/mL,which was lower than that before the intervention and had statistical significance(P <0.05).The AMH concentration in the experimental group A and experimental group B after the intervention was478.48±44.66pg/mL,510.48±54.38pg/mL,which was lower than that before the intervention,and the difference was statistically significant(P <0.05).The blood AMH after intervention in the elderly experimental group was 480.59±28.47pg/mL,which was significantly different from that before intervention(P <0.05).4.The results of ovarian tissue HE staining showed that the number of original follicles in the model control group and the elderly control group were 17.2±1.9 and16.4±1.14,and the number of growing follicles was 27±4.6 and 33.4±1.1,respectively.The difference was significant(P <0.05).The number of atretic follicles in the model control group and the elderly control group were 10.2±0.84 and 9.6±1.82,respectively,which was significantly increased compared with the blank control group(P <0.05).The number of follicles growing in experimental group A,experimental group B,and elderly experimental group were 40.2±4.1,43.2±4.8,and33.4±1.1,which were significantly increased compared with the model control group and the elderly control group.The difference was statistically significant.(P <0.05),the number of atretic follicles in experimental group A,experimental group B and elderly experimental group were 8.0±1.5,7.2 ±1.6,7.0±1.5,respectively,comparedwith the model control group and the elderly control group Significantly reduced,and the difference was statistically significant(P <0.05).The number of follicles growing in experimental group A and experimental group B were 40.2±4.1 and 43.2±4.8,which were significantly increased compared with the elderly experimental group,and the difference was statistically significant(P <0.05).The number of atretic follicles in experimental group A was 8.0 ±1.5,which was not statistically significant compared with experimental group B(P> 0.05).The number of growing follicles in experimental group A was 40.2±4.1,which was not different from experimental group B.Statistical significance(P> 0.05).5.The results of immunohistochemistry showed that the expression of YAP protein in the nucleus of granule cells of ovarian cortex of mechanically stimulated mice increased,and the difference was statistically significant(P <0.05).conclusion: 1.Mechanical stimulation of mouse ovarian tissue can activate dormant follicles in premature ovarian failure mice.2.Mechanical stimulation of premature senile ovaries may be caused by blocking the hippo signaling pathway and increasing the expression of YAP protein in the nucleus of granular cells. |
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