In vitro changes in porcine ovarian granulosa cells induced by copper

• Published in: Journal of environmental science and health. Part A, Toxic/hazardous substances & environmental engineering Toxic/hazardous substances & environmental engineering, 2014-05, Vol.49 (6), p.625-633
• Main Authors: Roychoudhury, Shubhadeep, Bulla, Jozef, Sirotkin, Alexander V., Kolesarova, Adriana
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Biological and medical sciences; Cell Proliferation - drug effects; Cells; Chemical and industrial products toxicology. Toxic occupational diseases; Copper; Copper - metabolism; Copper - toxicity; Cyclin B1 - metabolism; Dose-Response Relationship, Drug; Female; Granulosa Cells - drug effects; Growth factors; Humans; Immunohistochemistry; In Situ Nick-End Labeling; In vitro testing; Insulin-Like Growth Factor I - metabolism; Insulin-like growth factors; Markers; Medical sciences; Metals and various inorganic compounds; ovarian granulosa cells; Peptides; porcine; Progesterone - metabolism; Radioimmunoassay; secretion activity; Secretions; Steroids; Swine; Toxic; Toxicology
• ISSN: 1093-4529; 1532-4117
• DOI: 10.1080/10934529.2014.865404

Objective of this in vitro study was to examine the secretion activity (progesterone and insulin-like growth factor I) of porcine ovarian granulosa cells after copper (Cu) addition and to outline a potential intracellular mediator (cyclin B1) of its effects. It also aimed at investigating the apoptotic potential of Cu on porcine ovarian granulosa cells after addition in vitro. Ovarian granulosa cells were incubated with copper sulphate (CuSO 4 ·5H 2 O) at the doses 0.33, 0.40, 0.50, 1.0 and 2.0 μL mL −1 for 18 h and compared with control group without Cu addition. Release of progesterone (P 4 ) and insulin-like growth factor I (IGF-I) by granulosa cells was assessed by RIA, expression of cyclin B1 by immunocytochemistry and apoptosis by TUNEL assay. Observations show that P 4 release by granulosa cells was inhibited while the release of IGF-I and cyclin B1 was stimulated significantly (P < 0.05) by CuSO 4 ·5H 2 O addition at the dose 2.0 μL mL −1 . Also, addition of CuSO 4 .5H 2 O at the lowest dose used in the study (0.33 μL mL −1 ) significantly (P < 0.05) decreased apoptosis in granulosa cells. In conclusion, results indicate dose dependent effect of Cu on (1) secretion of steroid hormone progesterone and growth factor IGF-I, (2) expression of cyclin B1 as marker of proliferation of porcine ovarian granulosa cells, (3) apoptosis of porcine ovarian granulosa cells and, (4) that the effect of Cu on ovarian cell proliferation could be mediated by IGF-I and cyclin B1. Obtained data suggest interference of Cu in the pathways of proliferation of porcine ovarian granulosa cells through hormonal and intracellular peptide cyclin B1.