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.