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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.

Objective: This study aimed to investigate the enzymatic anti-oxidative, hemolytic and cytotoxic activities of crude mucus proteins from the two earthworms including Eudrilus eugeniae (African night crawler) and Perionyx excavatus (Blue worm)., Materials and Methods: The bioactivities were determined by hemolytic activity, cytotoxic activity using HepG2 human hepatocellular carcinoma cells, L-929 mouse fibroblast and TK6 human lymphoblast cell lines and antioxidant activity., Results: The results indicated that the hemolytic activity of mucus proteins of P. excavatus was higher than that of E. eugeniae. The cytotoxic activity of the mucus proteins of E. eugeniae could inhibit the growth of HepG2 with the IC50 at 144.2 ± 0.18 µg/mL but showed no effect on L-929 and TK6. On the other hand, the crude proteins of P. excavatus decreased cell viability of both L-929 and HepG2 with their IC50 respectively were 6.87 ± 0.16 and 174.3 ± 0.19 µg/mL but they did not reduce the growth of TK6 cell line. The SOD-like activity of P. excavatus and E. eugeniae crude proteins were found with the IC50 at 149 µg/mL and 386.2 µg/mL, respectively. For GPx-like activity, crude proteins of P. excavatus exhibited significant (p ≤ 0.05) greater activity than those of E. eugeniae when tested at the concentration 100 µg/mL., Conclusions: This study revealed the bioactivities of crude mucus of earthworms which served as alternative natural proteins for the prophylaxis or treatment of free radical-related diseases as well as development of dietary supplements and cosmetics.