The Effects Of Hippo Signaling Pathway In Mediating The Function Of Ovarian Exposured To Tri-Ortho-Cresyl Phosphate

Objection:In daily life,reproductive damage issues caused by the application of the plasticizer have caused the wide attention of the society.This study is aimed at exploring the effects of Hippo signaling pathway in regulating the function of ovarian and the initiation of primordial follicles exposured to Tri-ortho-cresyl Phosphate(TOCP)from the level of acute and chronic exposure.Methods:1.To analyze the reproductive toxicities of TOCP and the correlation between the core component of Hippo pathway and TOCP by determining serum hormone,coefficient of ovarian weight,morphological changes of ovarian,follicle counting,apoptosis of ovarian follicles and the expression changes of the core components of Hippo signaling pathway.2.The TOCP culture model of mice primordial follicle in vitro was established and HE staining,TUNEL staining and Western Blotting were used to analyze the reproductive toxicities of TOCP to the development of mice primordial follicle and the expression changes of the core components of Hippo signaling pathway.3.The mice model of TOCP-YAP1 was established and the coefficient of ovarian weight,morphological changes of ovarian,follicle counting and apoptosis of ovarian follicles were detected to research the role of Hippo-YAP1 in regulating the function of mice ovaries exposured to TOCP.4.The TOCP-YAP1 culture model of mice primordial follicle in vitro wasestablished and HE staining and immunofluorescence staining were used to analyze the effects of Hippo-YAP1 in the development of mice primordial follicle.Results:1.The coefficient of ovarian weight of mice exposured to TOCP reduced in dose dependent type(p<0.05,p<0.01).The levels of serum progesterone were decreased significantly(p<0.05,p<0.01).However,the levels of serum estradiol were statistically significantly higher(p<0.05).The results of HE staining showed that the numbers of primordial and mature follicles were reduced during TOCP-treated mice,but atretic follicles increased gradually.The TUNEL staining analyses showed that TOCP exposure increased apoptosis of ovarian follicles.The results of Western Blotting showed that compared with control group,the expression of MST1 and LATS2 and the ratio of BAX/BCL-2 were increased significantly(p<0.05,p<0.01,p<0.001),while the expression of YAP1 was decreased significantly(p<0.05,p<0.01).Moreover,the ratio of P-YAP1/YAP1 was increased(p<0.01).2.After cultured with TOCP 8 days in vitro,the results of HE staining showed that the structure of mice ovarian tissue was destroyed and follicle morphology was unusual and the total number of follicles was significantly reduced.The TUNEL staining analyses showed that TOCP exposure increased apoptosis of ovarian follicles.The results of Western Blotting showed that compared with control group,the expression of LATS2 were increased significantly in TOCP treated groups(p<0.05,p<0.01,p<0.001),while the expression of YAP1 was decreased significantly(p<0.05,p<0.01).Moreover,the ratio of P-YAP1/YAP1 were increased(p<0.05,p<0.01).3.The coefficient of ovarian weight of mice treated with TOCP-YAP1 increased(p<0.01).The results of HE staining showed that plenty of primordial follicles were activated and developed into primary follicles.The results of Western Blotting showed that compared with control group,the ratio of BAX/BCL-2 was declined(p<0.001).4.The HE staining showed that the total number of ovarian follicles transfected with high-expression lentiviral vector YAP1 in vitro was increased and plenty of primordial follicles were activated and developed into primary follicles.Conclusions:1.TOCP was proved to have high toxicity effect on the reproductive system of mice.2.The abnormal expression of Hippo signaling pathway is one of the pathological mechanisms of the female reproductive injury cased by TOCP.