The Role Of SKP2 And TFEB In Decidualization Of The Endometrium In Early Pregnancy Mice

PART 1 EFFECT OF S-PHASE KINASE-ASSOCIATED PROTEIN2 ON ENDOMETRIAL DECIDUALIZATION IN EARLY PREGNANT MICEObjective: The dynamic balance of proliferation and apoptosis of decidual cells is the key to maintaining normal decidualization.Skp2 acts as an important regulator of cell transition from G1 to S phase.Skp2 can ubiquitinate Cyclin,CKI and other target proteins,thereby regulating the cell proliferation and apoptosis.Studies have found that Skp2 plays an important role in the development of various cancers,but its role in the endometrial decidualization of early pregnant mice is still unclear.This study is aim to investigate the role of Skp2 in the decidualization of endometrium in early pregnant mice.Methods:1.Normal pregnancy model and pseudopregnancy model of early pregnant mice were constructed.The expression of Skp2 was detect by immunohistochemistry and Western blot.2.Artificial decidualization model in early pregnant mice was constructed.The expression of Skp2 was detected before and after decidualization by immunohistochemistry and Western blot.3.The decidualization induction model of primary mouse endometrial stromal cells was constructed and the expression of Skp2 was detected by Western blot.4.Construct a model of decidualization of primary mouse endometrial stromal cells overexpressing Skp2.The protein expression of decidual markers BMP2,MMP2,HOXA10 and Skp2 were detected in mouse endometrial stromal cells by Western blot.Apoptosis was detected by Flow Cytometry.Results:1.Western blot and immunohistochemistry showed that with the increase of days during pregnancy,the expression of Skp2 in D5,D6 and D7 was significantly lower than that in D1,and the protein expression of Skp2 in D5,D6,D7 endometrial implantation sites tissues were significantly lower than that of D5,D6,D7 endometrial inter-implantation site tissues.2.Western blot and immunohistochemistry showed that the expression of Skp2 in PD4 was significantly higher than that in PD6 and PD7.3.Western blot and immunohistochemistry showed that the expression of Skp2 protein in stimulated side was significantly lower than in unstimulated side in vivo artificial induced decidualization model.4.Western blot analysis showed that the expression of Skp2 protein in the primary mouse endometrial stromal cells was significantly lower than that in the uninduced group.After up-regulating the expression of Skp2,the expression of the decidual markers BMP2,MMP2 and HOXA10 was disordered.Flow cytometry results showed that apoptosis was significantly increased after up-regulation of Skp2 expression.Conclusion:This study preliminarily revealed that Skp2 may play an important role in the endometrial decidualization in early pregnant mice,and its specific mechanism needs further study.PART 2 EFFECT OF TRANSCRIPTION FACTOR EB ON ENDOMETRIAL DECIDUALIZATION IN EARLY PREGNANT MICEObjective: TFEB can affect apoptosis in many physiological and pathological processes by regulating the autophagy-lysosomal pathway.In the process of decidualization,the proliferation and degradation of decidual cells coexist.The two coordinate the number of decidual cells and the erosion of trophoblast cells in the dynamic balance,and maintain the normalization of decidualization.The research team found that FOXO3 a may participate in the endometrial decidualization of early pregnancy through the apoptotic pathway.As a downstream molecule that can be negatively regulated by FOXO3 a,it is unclear whether it is also involved in the endometrial decidualization in early pregnancy.Therefore,this study aimed to observe the expression of TFEB in the endometrium of mice during the peri-implantation period,and to explore the role of TFEB in the endometrial decidualization of early pregnant mice.Methods:1.Normal pregnancy model and pseudopregnancy model of early pregnant mice were constructed.The expression of TFEB was detect by q PCR and Western blot.2.Artificial decidualization model in early pregnant mice was constructed.The expression of TFEB was detected before and after decidualization by q PCR and Western blot.3.The decidualization induction model of primary mouse endometrial stromal cells was constructed and the expression of TFEB was detected by q PCR and Western blot.Results:1.The results showed that the expression of TFEB on D4,D5 and D6 gradually decreased with the increase of the number of days during pregnancy.Western blot,Real-time PCR and immunohistochemistry showed that the m RNA and protein expression of TFEB in D5 endometrial implantation site was significantly lower than that in D5 endometrial implantation tissue.2.Western blot and Real-time PCR showed that the expression of TFEB m RNA and protein in stimulated side was significantly lower than in unstimulated side.3.Western blot and Real-time PCR showed that the expression of TFEB m RNA and protein in the endometrial stromal cells in induce group was significantly lower than that in the control group.Conclusion:This study initially revealed that TFEB may be involved in the decidualization of endometrium in early pregnancy in mice,but its specific mechanism needs further study.