虾青素对超低温冷冻大黄鱼精子具有细胞膜保护作用

Membrane Protection Effect of Astaxanthin on Cryopreserved Sperms of Larimichthys crocea

为研究虾青素对冷冻损伤的大黄鱼精子细胞的保护作用,探讨其内在机制,我们将大黄鱼精子稀释于含10%二甲基亚砜和各浓度虾青素的冷冻稀释液中,冷冻并解冻后,显微镜观察发现,1.43×10-4mol/L的虾青素使大黄鱼精子寿命、运动时间和激活率分别增加13.8%、36.0%和5.5%,接近于鲜精;单细胞凝胶电泳显示,核DNA受损程度明显减少,彗星拖尾变短;流式细胞术结果显示,质膜和线粒体明显受到保护,线粒体质量提高了19.27%(P<0.05)。通过双层类脂膜法测定不同浓度虾青素对H+的传递能力,发现2%的虾青素能较高效率地传递H+。结果表明适当浓度的虾青素能通过传递H+维持线粒体功能,从而保护冷冻损伤的大黄鱼精细胞。

This study was designed to determine the protective effects of astaxanthin on the frozen-thawed impaired larimichthys crocea sperm and reveal its possible mechanisms. The sperm life time, moving time and activation ratio of thawed sperm observed by microscopy showed the astaxanthin of 1.43x10^-4 mol/L concentration increased the sperm activity. Single cell gel electrophoresis technique （SCGE） showed damaged nuclei DNA reduced and the comet tail shortened significantly （P〈0.05）. Flow cytometry showed the ratio of both intact plasma membrane and normal mitochondrial of thawed sperm increased by 19.27% （P〈 0.05）. The proton transferring function of astaxanthin was detected by bilayer lipid membrane （BLM） method. Astaxanthin at the ratio of 2% could transfer H＋ efficiently. This revealed that appropriate concentration of astaxanthin might exert its protection on impaired ceils via its proton transferring function.