Effect And Mechanism Of GnRH-agoniston Adenomyosis Therapy

The pathological features of adenomyosis are invasion of the endometrium into the myometrium and ectopic endometrial glands surrounded by hypertrophic and hyperplastic myometrium.With the wide clinical application of the ultrasonography and magnetic resonance inspection,especially with the usage of transvaginal ultrasonography,early detection and timely treatment of adenomyosis have been realized.Recent studies have showed that adenomyosis and infertility are closely related.The poor motility of embryo implantation,human sperm and poor uterus contractionsare are linked to infertility for adenomyosis patients.In addition,adenomyosismay decrease the pregnancy rate and raise abortion rates in IVF.GnRH agonist is a drug used in the treatment of adenomyosis.Conventional IVF usually used long contral ovarian stimulation.It can suppresses ovulationand then improve the endometrial receptivity.But it also has some drawbacks,such as OHSS risks and less number of eggs retrieved.We first propose“two-step approach” for the treatment of adenomyosis.Whether GnRH agonist treatment before FET can improve the pregnancy outcome,there is less research on it.In addition,the mechanisms of GnRH agonists are poorly understood.The research of this subject is to study the effect and mechanism of GnRH-agonist.The issue was consists of two parts: first one was to research the effect of GnRH agonist on adenomyosis,and the second one was to research the mechanisms of GnRH agonists on adenomyosis.The first one was a retrospective analysis of 139 patients who were diagnosed with infertilitybetween Jan 2011 and Dec 2014.The cycles were divided into groups A,B and C,according to different treatment methods before FET protocol.Group A consisted of non-adenomyosis patients who received pure hormone replacement therapy(HRT).Group B consisted of adenomyosis patients who received HRT.Group C consisted of adenomyosis patients who received GnRH agonist therapy and HRT.The patients' medical records were reviewed,including pregnancy outcomes and neonatal outcomes.The clinical data did not differ singnificantly among the three groups.The implantation rate(33.8%vs.25.0%,p>0.05)and live birth rate(44.7%vs.25.0%,p>0.05)in group C were higher than those in group B.The miscarriage rate(6.6%vs.6.8%,p>0.05)in group C was lower than those in group B.Our study showed that among the adenomyosis patients,Group C had a significantly higher live birth rate than Group B(44.74% vs.25%,P = 0.034).Neonatal outcomes,including birth weight,sex,birth defects,singleton vs.twins and mode of delivery,were compared.No differences were observed among the groups.GnRH agonist therapy before FET may offer a higher live birth rate in patients with adenomyosis.Additionally,the treatment itself appears not to increase birth defect risk.In part two,high-throughput sequencing was used to explore the mechanisms of GnRH agonist on adenomyosis.Finally,we explored the relation between autophagy,apoptosis and GnRH agonist.It is difficult to obtain endometrial samples from women undergoing such treatment.In order to overcome this flaw,we generated an adenomyosis mouse model.Neonatal mouse were fed with tamoxifen to modeling.Neonatal female mice were divided into a control group,an untreated adenomyosis group,and an adenomyosis group treated with a GnRH agonist.Randomly chosen transcriptomes from endometrial tissues from day 4 of pregnancy were analyzed between the adenomyosis group and the GnRH agonist treatment group byRNA sequencing.Three hundred and fifty-nine genes were differentially expressed in the GnRH agonist-treated group compared with the untreated group(218 were downregulated and 141 were upregulated).Differentially expressed genes were related to diverse biological processes,including estrogen metabolism(Tff1?Sprr2a1?Arnt2?Cxcl2),cell cycle(Tff1?Wnt11?Msx1).Go and Pathwayfunctional analysis further validated the results.The results suggested that besides pituitary down-regulation,other possible mechanisms such as the regulation ofcell cycle may play a role in this.Transmission electron microscope(TEM)was used to observe autophagosome.Western-blot were used to investigate the expression of autophagy related proteins(Beclin 1,LC-3).Apoptotic level of endometrial cell was evaluated by TUNEL method.Cell in the model group showed significantly fewer autophagosomes and less Beclin 1,LC-3 expression in the eutopic endometrium than control mice.However,in model mice that received a GnRH agonist,significantly more autophagosomes and Beclin 1,LC-3 protein expression were observed than the model mice that received vehicle.These results indicate that adenomyosis may cause inhibition of autophagy level,but that is maybe reversed by GnRH agonists.There was no significant difference in endometrial cell apoptotic rate among the three groups.The study suggested that targeting aberrant autophagy might be a novel mechanism of GnRH agonist for the treatment of adenomyosis.We suspect that GnRH agonist can improve endometrial receptivity to some extent,and then increase pregnancy rate.In conclusion,the results validated the safety and efficacy of “two-step approach” in the treatment of adenomyosis,it supported the clinical application of GnRH agonist.We also discovered the mechanism of adjust cell cycle in the treatment of adenomyosis.Simultaneously,we found the relation between GnRH agonist and autophagy.Therefore,this study settled the base for future research.