Luzindole and 4P-PDOT block the effect of melatonin on bovine granulosa cell apoptosis and cell cycle depending on its concentration

Abstract

Granulosa cells play an essential physiological role in mediating the follicle development and survival or apoptosis of granulosa cells dictate the follicle development or atresia. The aim of this study was to investigate the role of high dose (10−5 M) and low dose (10−9 M) melatonin in bovine granulosa cells, and assess whether MT1 and MT2 inhibiter affect granulosa cells response to melatonin. We found that the high dose (10−5 M) and low dose (10−9 M) both could act as an essential role in modulating granulosa cells apoptosis, cell cycle and antioxidant. The beneficial effect could be related to that melatonin promoted the expression of Bcl2, Bcl-xl, SOD1 and GPX4, and inhibited Bax, caspase-3 and p53 expression. Moreover P21 expression was decreased in granulosa cells treated with the high dose (10−5 M) melatonin and increased in that treated with the low dose (10−9 M) melatonin. To further reveal the role of MT1 and MT2 in mediating the effect of melatonin on granulosa cells apoptosis, cell cycle and antioxidant, we found that the luzindole and 4P-PDOT did not affect the effect of high dose (10−5 M) melatonin on regulating Bcl2, Bax, caspase-3, SOD1, GPX4 and p53 expression, while blocked its effect on modulating Bcl-xl and P21expression. However, luzindole and 4P-PDOT disturbed the effect of low dose (10−9 M) melatonin on regulating Bcl2, Bax, caspase-3, Bcl-xl, SOD1, GPX4, and p53 expression. In conclusion, these results reveal that the effect of low dose (10−9 M) melatonin on granulosa cells apoptosis are mediated by MT1 and MT2, and the high dose (10−5 M) melatonin affect the granulosa cells apoptosis by other pathway, besides MT1 and MT2. Moreover MT1 and MT2 may work in concert to modulate bovine granulosa cells function by regulating cellular progression and apoptosis.

The genomic response of human granulosa cells (KGN) to melatonin and specific agonists/antagonists to the melatonin receptors

“…Researchers have shown that the two MSI proteins play critical roles in the proliferation and differentiation of stem cells of the hematopoietic and nervous systems 121,122 . Nonetheless, Wang and al., 2021 confirmed that MT2 inactivation in bovine GC by 4P-PDOT may affect the regulatory action of melatonin on GC functions 123 .…”

Melatonin Type 2 Receptor Activation Regulates Blue Light Exposure-Induced Mouse Corneal Epithelial Damage by Modulating Impaired Autophagy and Apoptosis

“…Although MT2 activation promotes apoptosis under BL-induced stress, our current data indicate that MT2 induction protects corneal epithelial cells from impaired autophagy and ROS stress. Many previous studies have demonstrated that apoptosis is an important pathway that protects cells from damage, as the balance between BCL2 and BAX can protect mitochondrial function and proliferation in normal cells and other tissues [ 4 , 46 , 47 , 48 , 49 , 50 ].…”

Regulatory role of melatonin on epidermal growth factor receptor, Type I collagen α1 chain, and caveolin 1 in granulosa cells of sheep antral follicles

“…Several studies have shown that MLT receptors are expressed in the ovaries of various mammals, including sheep and that the function of MLT in mammalian ovaries is mainly mediated by MT1 and MT2 (He, Deng, et al, 2016; Lee et al, 2018; Tian et al, 2017; Yang et al, 2017). It has also been suggested that the MT1 receptor and EGF/EGFR‐mediated signaling pathway may interact significantly during cell proliferation because the MT1 receptor activates MLT antiproliferation signaling by mediating EGF/EGFR signaling (Wang et al, 2021). However, there are some differences between the results of these studies and our results, which may be attributed to different types and sources of melatonin and the fact that there are species differences in the response of granulosa cells to EGF (Wang et al, 2012).…”