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Účinky xenobiotik na oxidační stres, metabolizmus lipidů, integritu DNA a životaschopnost lidských buněk a rybích spermií in vitro
LINHARTOVÁ, Pavla
Pollution of the aquatic environment by inorganic and organic chemicals is a major factor posing a serious threat to the survival of aquatic organisms including fish. In addition balancing risks and benefits of fish consumption is nowadays an intensively discussed public health topic. Spermatozoa of almost all fish species are released into water environment where they can be directly exposed to various compounds, such as xenobiotics including toxic metals, prior to fertilization. In addition, exposure of parental adults to various xenobiotics may affect gamete quality, which may subsequently reduce fertilization success. On the other hand the advantages of eating fish are well-known, not only in the point that fish is a healthy source of protein and other nutrients, but eating contaminated fish may also confer various health benefits. Research over the past few decades has shown that the nutrients and particularly the n-3 fatty acids (FA) found in fish and seafood, are for examples protective against cardiac diseases and have a positive impact on brain development. The thesis provides a focus on two different cell model types. Firstly, human hepatocellular cells (Hep G2, ATCC) were used as in vitro tool for studying the effect of the intake of cadmium (Cd2+) contaminated fish on cytotoxicity, oxidative stress and fatty acid and phospholipid class compositions. Secondly, spermatozoa of one threatened species of fish, sterlet (Acipenser ruthenus) were used as in vitro model for studying effect of potentially hazardous xenobiotic compounds' occurring in open waters. Sperm from sterlet were exposed for 2h to environmentally relevant concentrations of DQ (0-150
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Efekt xenobiotik na DNA integritu a fyziologii rybích spermií
LINHARTOVÁ, Pavla
Pollution of the aquatic environment with xenobiotics has become a serious health concern in recent years. In the present study the effect of DQ, TBBPA, BPA and VIN on sperm quality parameters, DNA integrity and oxidative stress indices in sterlet (Acipenser ruthenus) sperm and sperm from brook trout (Salvenilus fontinalis) were investigated. To do this, an in vitro spermatozoa motility assay was used by a computer-aided Motion-Analysis system. The sperm of sterlet (Acipenser ruthenus) was diluted to obtain the spermatozoa density of 5×108 cells×ml?1 and then exposed for 2 h to final concentrations of xenobiotics: DQ - 25, 50, 100 and 150 ?M, TBBPA - 0.5, 1.75, 2.5, 5, 7.5 and 10 ?g/l, BPA - 0.5, 1.75, 2.5, 5 and 10 ?g/l and Vin - 0.5, 1.75, 2.5, 5 and 10 ?g/l. Spermatozoa velocity and percentage of motile sperm were significantly decreased at each time post-activation compare to control. The level of DNA damage expressed as a % DNA in Tail and Olive Tail moment significantly increased when spermatozoa were exposed to higher concentrations of xenobiotics. The level of oxidative stress indices lipid peroxidation (LPO) and carbonyl derivatives of proteins (CP) and antioxidant activity of superoxide dismutase (SOD) increased significantly with increasing concentration of xenobiotics. On the other hand the intracellular ATP content in sperm samples had a significantly decreasing effect. In short, xenobiotics can induce reactive oxygen species (ROS) stress in fish spermatozoa, which could impair the sperm DNA integrity, quality and antioxidant defense system. The present study confirms that environmental concentrations of xenobiotics are capable to induce oxidative stress, leading to impaired sperm quality, DNA fragmentation and intracellular ATP content Obtained results also suggested that the use of fish spermatozoa in vitro assays may provide a novel and efficiently means for monitoring residual pharmaceutical in aquatic environment.
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