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Antioxidative Effects of Astaxanthin against Nitric Oxide‐Induced Oxidative Stress on Cell Viability and Gene Expression in Bovine Oviduct Epithelial Cell and the Developmental Competence of Bovine IVM/IVF Embryos
Author(s) -
Jang HY,
Ji SJ,
Kim YH,
Lee HY,
Shin JS,
Cheong HT,
Kim JT,
Park IC,
Kong HS,
Park CK,
Yang BK
Publication year - 2010
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/j.1439-0531.2009.01469.x
Subject(s) - astaxanthin , andrology , oxidative stress , viability assay , in vitro maturation , sodium nitroprusside , nitric oxide , lipid peroxidation , chemistry , biology , apoptosis , microbiology and biotechnology , embryo , endocrinology , biochemistry , oocyte , medicine , carotenoid
Contents The aim of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co‐culture on developmental capacity of bovine in vitro oocyte maturation/ in vitro fertilization (IVM/IVF) embryos. We examined the effects of astaxanthin against nitric oxide‐induced oxidative stress on cell viability by MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and the expression of antioxidant genes ( CuZnSOD, MnSOD and Catalase ) or apoptosis genes ( Bcl‐2, Caspase‐3 and Bax ) by RT‐PCR in BOEC. We also evaluated the developmental rates of bovine IVM/IVF embryos co‐cultured with BOEC pre‐treated with astaxanthin (500 μ m ) in the presence or absence of sodium nitroprusside (SNP, 1000 μ m ) for 24 h. Cell viability in BOEC treated with SNP (50–2000 μ m ) lowered, while astaxanthin addition (50–500 μ m ) increased it in a dose‐dependent manner. Cell viability in astaxanthin plus SNP (1000 μ m ) gradually recovered according to the increase in astaxanthin additions (100–500 m m ). The LPO in astaxanthin group (50–500 μM) gradually decreased in a dose dependent manner and among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis or antioxidant genes was detected by RT‐PCR. Bcl‐2 and antioxidant genes were detected in astaxanthin or astaxanthin plus SNP group, and Caspase‐3 and Bax genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 6–7 days under co‐culture system such as BOEC treated with astaxanthin in the presence or absence of SNP, the developmental ability to blastocysts in 500 μ m astaxanthin group was the highest of all groups. These results suggest that astaxanthin has a antioxidative effect on cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis genes.