| This study was designed to determine the protective effects of astaxanthin on the impaired cells and reveal its possible mechanisms.Hepatocyte L-02 cells were exposed to 100 mmol/L NaN3 with various concentrations of astaxanthin pre-incubated. MTT assay showed the cell viability was increased. Changes of mitochondrial membrane potential (MMP) and apoptosis ratio detected by JC-1 method and Annexin V-FITC/PI double stain method, respectively, were found the MMP was maintained and the late phase apoptosis ratio reduced. However, the level of reactive oxygen species determined by DCFH-DA method showed insignificant decrease. The beneficial concentration of astaxanthin in cell viability and MMP were not in a dose dependent manner and the most effective of which was 0.10 nmol/L (P<0.01).In addition, the protective effects of astaxanthin on frozen-thawed larimichthys crocea sperm were determined. The sperm life time, moving time and activation ratio of thawed sperm observed by microscopy showed the concentration of 1.43×10-4 mol/L astaxanthin 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). To study the potential non-antioxidant mechanisms of astaxanthin in our experiment, the prone transferring function of astaxanthin was detected by bilayer lipid membrane (BLM) method. Results showed that astaxanthin at the ratio of 2.0 % could transfer H+ efficiently. This revealed that appropriate concentration of astaxanthin might have some proton pressure and transfer H+, by which the MMP could be maintained.To sum up, the mechanisms of astaxanthin in protection of impaired cells might via its proton transferring function, by which improved the mitochondrial function and had the sequence biology effects.
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