Protective effects of melatonin against hypoxia-induced TM3 cell damage via suppression of the TGF-β pathway

Rashidi, Sahar; Saberivand, Adel; Gholami, Mohammadreza; Asadollahi, Vahideh; Alasvand, Masoud; Saberivand, Maryam; Ghasemi, Mona

doi:10.30466/vrf.2024.2039638.4415

Protective effects of melatonin against hypoxia-induced TM3 cell damage via suppression of the TGF-β pathway

Document Type : Original Article

Authors

Sahar Rashidi ¹

Adel Saberivand ¹

Mohammadreza Gholami ²

Vahideh Asadollahi ³

Masoud Alasvand ⁴

Maryam Saberivand ⁵

Mona Ghasemi ¹

¹ Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

² Medical Technology Research Center, Institute of Health Technology, Kermanshah University of Medical Sciences, Kermanshah, Iran

³ Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran

⁴ Cancer and Immunology Research Center, Research Institute for Health Development, Kurdistan University of Medical Science, Sanandaj, Iran

⁵ Connective Tissue Diseases Research Center, Tabriz University of Medical Science, Tabriz, Iran

10.30466/vrf.2024.2039638.4415

Abstract

This study aimed to explore whether melatonin protects TM3 Leydig cells from cobalt (II) chloride (CoCl₂)-induced hypoxia through the transforming growth factor beta (TGF-β) signaling pathway. Cells were divided into four groups: a control group without treatment (Group 1), a melatonin group (10.00 ng mL-1; Group 2), a group treated with CoCl2 (100 µM) to induce hypoxia (Group 3), and a melatonin + CoCl₂ group (Group 4). After 96 hr of incubation, cell viability was assessed using the MTT assay, and transforming growth factor beta 1, activin receptor-like kinase-5, and bone morphogenetic protein 4 gene and protein expressions were measured through RT‑PCR and western blotting. The CoCl₂ and melatonin + CoCl₂ groups exhibited significantly diminished cell viability compared to the control. However, melatonin treatment enhanced survival in the CoCl₂-exposed cells. Notably, transforming growth factor beta 1 expression was elevated in all groups. Activin receptor-like kinase-5 (gene and protein expression increased in CoCl₂-treated groups but was lower in the melatonin + CoCl₂ group. Melatonin treatment reduced bone morphogenetic protein 4 expression compared to the control, while CoCl₂ groups showed increased bone morphogenetic protein 4 levels. These findings suggest melatonin's potential as a therapeutic agent against oxidative stress and hypoxia in TM3 cells through its antioxidant properties.

Keywords

Cobalt (II) chloride

Leydig cells

Melatonin

TGF-&beta

signaling pathway