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Cellular enzymatic anti-oxidants of fractionated mucus proteins from Eudrilus eugeniae (African night crawler) and Perionyx excavatus (Blue worm) in MC3T3.

OBJECTIVE
To investigate SOD-liked, GPx-liked and cellular anti-oxidation activities of fractionated mucus proteins from Perionyx excavatus (Pe) and Eudrilus eugeniae (Ee) in the MC3T3 osteoblast precursor cell line.


MATERIALS AND METHODS
The crude mucus proteins were extracted from Pe and Ee and fractionated using the anion exchange chromatography method with FPLC. The fractionated proteins were studied to determine their SOD-liked and GPx-liked activities. The most efficient fractions were studied for cellular anti-oxidation activities in the MC3T3 cell line including scavenging, protecting and repairing conditions.


RESULTS
The results showed that the highest SOD-liked activities of EeANX1 were found at 4.17 µg/ml and GPx-liked activities of EeANX4 were found at 5.26 µg/mL. EeANX1 had no cytotoxic effect on MC3T3 at 500 µg/mL, and the IC50 of EeANX4 was over 300 µg/mL. When both EeANX1 and EeANX4 were investigated for cellular anti-oxidation activity in MC3T3 cells at 20 µg/mL, the cellular superoxide and total ROS production of were significantly (p<0.05) lower than those of 250 µM pyocyanin (ROS inducer) in ROS scavenging, protecting and repairing conditions. The 20 µg/mL of EeANX1 and EeANX4 demonstrated increased SOD and GPx activities in MC3T3. Investigation of the scavenging, protection and repairing conditions of MC3T3 treated with 20 µg/mL each of EaANX1, EaANX4 and 250 µM pyocyanin demonstrated that the proteins had significantly lower SOD activities and higher GPx activities than the 250 µM pyocyanin.


CONCLUSIONS
The EeANX1 and EeANX4 fractions demonstrate SOD-liked and GPx-liked activities, as well as cellular anti-oxidation activities. These fractions could be developed as a natural anti-oxidant. This research could provide benefit to the study of cellular anticancer.