Melatonin Attenuates Nitric Oxide Induced Oxidative Stress on Viability and Gene Expression in Bovine Oviduct Epithelial Cells, and Subsequently Increases Development of Bovine IVM/IVF Embryos

Kim, J. T.; Jang, H. Y.; Park, C. K.; Cheong, H. T.; Park, I. C.; Yang, B. K.

doi:2011.24.2.190

Article

Animal Reproduction and Physiology

Asian-Australasian Journal of Animal Sciences 2011;24(2): 190-197.
https://doi.org/10.5713/ajas.2011.10188 Published online December 21, 2010.

Melatonin Attenuates Nitric Oxide Induced Oxidative Stress on Viability and Gene Expression in Bovine Oviduct Epithelial Cells, and Subsequently Increases Development of Bovine IVM/IVF Embryos

J. T. Kim, H. Y. Jang, C. K. Park, H. T. Cheong, I. C. Park, B. K. Yang

Abstract

The objective of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co-culture on developmental capacity of bovine IVM/IVF embryos and to determine whether or not melatonin acts as an antioxidant in BOEC culture and subsequent embryo development. These studies examined the effects of melatonin against NO-induced oxidative stress on cell viability, lipid peroxidation (LPO) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl-2, Caspase-3 and Bax) during BOECs culture. We also evaluated the developmental rates of bovine IVM/IVF embryos with BOEC co-culture, which were pre-treated with melatonin (1,000 M) in the presence or absence of sodium nitroprusside (SNP, 1,000 M) for 24 h. Cell viability in BOECs treated with SNP (50-2,000 M) decreased while melatonin addition (1-1,000 M) increased viability in a dose-dependent manner. Cell viability in melatonin plus SNP (1,000 M) gradually recovered according to increasing melatonin addition (1-1,000 M). The LPO products were measured by thiobarbituric acid (TBA) reaction for malondialdehyde (MDA). Addition of melatonin in BOEC culture indicated a dose-dependent decrease of MDA, and in the SNP group among BOECs treated with SNP or melatonin plus SNP groups MDA was significantly increased compared with SNP plus melatonin groups (p<0.05). In expression of apoptosis or antioxidant genes detected by RT-PCR, Bcl-2 and antioxidant genes were detected in melatonin or melatonin plus SNP groups, while Caspase-3 and Bax genes were only found in the SNP group. When bovine IVM/IVF embryos were cultured for 6-7 days under the BOEC co-culture system pre-treated with melatonin in the presence or absence of SNP, the highest developmental ability to blastocysts was obtained in the 1,000 M melatonin group. These results suggest that melatonin has an anti-oxidative effect against NO-induced oxidative stress on cell viability of BOECs and on the developmental competence of bovine IVM/IVF embryo co-culture with BOEC.

Keywords: Melatonin; Bovine Oviduct Epithelial Cell; Bovine IVM/IVF Embryos; Nitric Oxide; Antioxidant