Effect Of CMKLR1 Deficiency On Ovarian Function In Mouse DHT-induced Like-PCOS Model

Polycystic ovary syndrome(PCOS),the most common endocrine disorder in women in reproductive age. Chemerin is newly found as a adipokines,play an important role in the process of immune response, inflammation, adipose differentiation, glycometabolism, angiogenesis and reproductive physiological process. The bone morphogenetic protein(BMP) family have some effect on estrogen and progesterone secretion, as well as reproduction. Given the role Chemerin and its receptor CMKLR1 in energy metabolism and reproductive function, its role in PCOS is becoming more and more attention. Therefore, the present study use DHT-induced like-PCOS mice model, compared the CMKLR1 knockout mice with the C57BL/6J wild-type mice,use HE staining、RT-qPCR、IF and ELISA to investigate the effect of CMKLR1 deficiency on ovarian function in mouse DHT-induced like-PCOS model.The following results were obtained:1.Hyperandrogenism induced like-PCOS model in wild type and CMKLR1-/- miceAfter 5α-DHT treated, the mice results in estrous cycle disorders, basically no estrus appear; abnormal ovarian morphology, showing polycystic changes, shrinkage of minimal large antral follicles, the absence of corpora lutea and ovulation disorder. Indicate the DHT-induced Like-PCOS model success.Compared with the DHT-treated wild type mice, the DHT-treated CMKLR1-/- mice were able to maintain 1-2 estrous cycle and certain numbers of follicles at different stages and corpora lutea. Indicate that CMKLR1 gene deletion can significantly reliever the DHT induced like-PCOS feature.2.The effect of CMKLR1 deficiency in ovarian function of DHT-induced like-PCOS model The mice were divided into control wild type mice(WT-CTRL), DHT-treated wild type mice(WT-DHT), and control CMKLR1-null mice(KO-CTRL), DHT-treated CMKLR1- null mice(KO-DHT). The results showed that:(1)By detecting the apoptosis of ovarian in four groups, found that compare with follicles in WT-DHT group, the KO-DHT exhibit weak apoptosis cell in theca cell and interstitial cell in wildtype mouse.(2)By detecting the serum estrogen, progesterone levels and steroid synthesis enzyme mRNA expression in four groups, DHT-treated CMKR1 null mice maintain a relatively high estradiol and progesterone level and more steroid synthesis enzyme than DHT-treated wildtype mice. These results indicate that, CMKLR1 deficiency protects against the DHT-induced ovarian dysfunction.3 The BMP4 signaling pathway may be involved in CMKLR1-dependent responses to DHTBy detecting the mRNA expression of BMPs and its receptor in four groups ovary, found that after DHT treated, BMP4 and Alk6 were significantly raised; compared with WT- DHT group, the BMP4 and ALK6 were significantly decreased in KO- DHT group,suggesting the BMP4 may be invoved in the CMKLR1 role in ovarian dysfunction induced by DHT.These results suggest that, CMKLR1 deficiency protects against the DHT-induced ovarian dysfunction,maintain the follicular development and the formation of corpus luteum, as well as the level of progesterone secretion, and partial recovery estrous cycle disorder caused by DHT, this effect may via BMP4 signaling pathway.