The Kisspeptin Concentration In Serum And The Expression Of KISS1,KISS1R Gene In Endometrium Of Patients With Repeated Implantation Failure

BackgroundRecurrent implantation failure(RIF), a kind of iatrogenic condition, is the result of repetitively unsuccessful cycles of IVF or intracytoplasmic sperm injection(ICSI) treatment.Most researches stated the definition of RIF as�three or more failed treatment cycles 'or �ten or mo re High quality embryos failed cycles. Life table analyses of cumulative pregnancy rates f oll owing multiple cycles of IVF have sho wn that assisted reprodu ction treatment is not the panacea for the couples who are still child less after multiple cycles of treatments. At present etiology of RIF is still uncertain. It is generally recognized that RIF is related to the decrease of Endometrial receptivity, abnormal heredity and devel opment of embryo, implantation techniques and immune factors.GPR54 is a G protein coupled recep tors, encoded by KISS1 R with seventransme mbrane domain structure, and which was found in 1997 by Lee[1] fr om the rat.In 2001, the cognate receptors of GPE54 we re found, named as h OT7T175 andAXOR12, and found that kisspeptin was the endogenous ligand of GPR54. Hypothalamic KISS1 and its derivatives(kisspeptins) are now well recognized as potent stimulators of Gn RH secretion and thereby major regulators of the neuroendocrine-reproductive axis.Diminished hypothalamic kisspeptin signaling in man and mouse, as a consequence of inactivating mutations in the genes that encode either KISS1 or its cognate receptor, KISS1 R, results in infertility.Suggests that Kisspeptin and its receptors in the Mice and human body is closely related to reproductive function.Janneau[3] et al. and Bilban[4] et.al confirmed respectively by RT-PCR and gene microarrays that KISS1 and KISS1 R are largely expressed in the human placenta and that their expression was much greater in the highly invasive early placenta compared to the noninvasive term placenta.it seemed likely that KISS1 plays an important role in regulating trophoblast invasion during early placentation and thus the placenta-based kisspeptin/KISS1 R signaling system is a regulator of human placentation.In addition,Calder [5] and other research have shown that transplant failure Kiss1-/- mothers cannot endometrial decidual, On the contrary, exogenous supplementary a reliable marker of the receptivity of endometrium leukemia necrosis factor(LIF) can improve the situation so as to achieve a successful pregnancy.it seemed likely that kisspeptin is not just a regulator of trophoblast function during early placentation but also a regulator of the preceding implantation event as well.Previous studies about KISS1 gene and its recep tor KISS1 R in the human body were mo re mainly f ocus on the central ne rvous system and ovary.In this study,we examined the expression of KISS1 gene in endometrium on the stage of proliferation and secretio n for the repeated implantati on failure patients and normal pregnancy patient s respectively. And as well as the c oncentration of protein Kisspeptin in serum of the two groups patients in the hyperplasia period a nd secretory period. Comparing Kisspeptin concentratio n in serum and th e m RNA expression of KISS1 gene in endometrium of this two groups of patients during proliferati on and secretion, we found that Kisspeptin concen tration in serum and the KI SS1 gene expression in endometrium of the control group patients on stage of proliferation are higher than that on stage of secretion. In addi tion, we also st udied the expression diff erences ofKISS1 gene and its receptors in endometrim among the RIF group, to figure outwhether the repeated transplant failure is related to the abnormal signaling pathwaysof KISS1.PurposeIn this study,we tested patients with repeated implantation failure and normalpregnancy patients KISS1 gene and KISS1 R gene expression in endometrium on thestage of proliferation(D11-13 days) and secretion(D20-25 days), examined theconcentration of protein Kisspeptin in serum of the two groups patients in theperiods of hyperplasia and secretory, compared the concentration of Kisspeptin inserum between these two groups during the periods of proliferation and secretion, aswell as the expression differences of m RNA of KISS-1 and KISS-1R in endometrim.Finally, we clarified whether repeated transplant failure is related to the abnormal ofKISS1 signaling pathways..Methods1. Collect 3 cases serum a nd endometrial of successful pregnancy patients and repeated implantation failure(RIF) patients respectively during hyperplasia period(D11-13 days) and secretory period(D20-25 days). Examine the concentration of E2, P in serum, and compare whether there is a diff erence between these two groups. Ta ke pathologic examinations to all of the endomet rium to identify themenstrual phases.2. Use enzyme-linked immuno sorbent assay(ELI SA) to determinate the concentratio n of Kisspeptin in serum of successful pregnancy patients and RIF patients on the stage of hyperplasiation(D11-13 days) and secretion(D20-25 days). Then compare the concentration of Kisspeptin in the two groups patients in two menstrual phases respectively.3. Measure the expression of KISS1 a nd KISS1 R by RT-PCR in successful pregnancy patients and repeated implantati on failure(RIF) patients on the stage of hyperplasiation(D11-13 days) and secreti on(D20-25 days). Co mpare the expression of KISS1 and KISS1R of two stages of me nstrual cycle in the same group, and the expression of KISS1 and KISS1R of t w o groups of patients.Results1. Comparing E2, P concentration in serum of two groups patients, and found that there was no statistical significance difference. The pathological examination showed that the endometrium of two groups of patients on the stage of hyperplasiation(D11-13 days) and secretion(D20-25 days) respectively presents the change on the stage of proliferation and secretion.2. In serum, two groups patients in hyperplasia period(D11-13 days) Kisspeptin concentration was higher than of that of secretory peroid(D20-25 days). In the control group, KISS1 m RNA expression in the endometrial hyperplasia stage(D11-13days) is higher than that in the secretory stage(D20-25 days). So we could conclude that the serum concentration and endometrium of Kisspeptin in cyclical changes.3. RT-P CR results are shown as follows:(1) The expression of KISS-1 m RNA during hyperlasia period is higher than that during secretory in the control group during menstrual phase, while there is no diff erence in PIF group during the same period; Hyperplasia of the control group(D11-13 days) and secretory(D20-25 days) KISS1 R m RNA expression is no difference. In RIF group secr etory(D20-25 days) KISS1 R m RNA expression was lower than that of hyperplasia(D11-13 days);(2) the comparison of two groups, Hyperplasia(D11-13 days) KISS 1R m RNA expression of two groups is no diff erence, but RIF group KI SS 1 m RNA expression is significantly higher than control group; Secretion(D20-25 days) KISS1 m RNA expression is no difference in two groups, but RIF group KI SS1 R m RNA expression is significantly reduced. Conclusion1. The Kisspeptin concentration in serum and the expression of KISS1 gene in endometrium of patients in normal pregnancy group change along with the menstrual cycle.2. Compared with hyperplasia, there is a significant reduction of the expression of KISS1 R gene in the secretory period of patients in RIF group.3. Compared with control group,during secretory there is a significant reduction of the expression of KISS1 R gene of patients with RIF.