To study reverse effect of the oxidative damage on cartilage cells of velvet antler polypeptides (VAPS), and to investigate the main mechanism of VAPS to protect cartilage cells through antioxidant.
Fifteen Japanese white rabbits of 5-month-old were selected in this study. Animal model was established by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut, articular cartilage cell cultured in vitro. Cells in the sham operation group was the normal control group, osteoarthritis cartilage cells in the model groups were added VAPS 6.25, 12.5, 25 microg/ml respectively. A group of animals were sacrificed every week form the ninth weeks(two months) and the cartilage cells were isolated and cultured. For 8 weeks,the reactive oxygen species level in chondrocytes were detected by DCFH-DA, the content of NO, SOD and GSH-Px in cell culture supernatant were detected by Griess method.
DCFH-DA detection of intracellular reactive oxygen species was (5.46 +/- 0.46)in the control group, (12.08 +/- 0.74) in the model groups. The model group compared with the control group by t test with the P value less than < 0.001. DCFH-DA detection of intracellular reactive oxygen species was (9.81 +/- 0.59)in VAPS 6.25 microg/ ml group, (7.83 +/- 0.63) in the VAPS 12.5 microg/ml group, (6.89 +/- 0.71) in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.05). The content of NaNO2, SOD and GSH-Px in osteoarthritis model group was (5.60 +/- 0.45) microM, (38.56 +/- 12.53) U/ml and (151.90 +/- 25.60) U, as compared with control group there were statistically significant difference (P < 0.001, P < 0.05); The content of NaNO2 was (4.34 +/- 0.39), M in VAPS 6.25 microg/ml group, (3.67 +/- 0.36) microM in the VAPS 12.5 microg/ml group, (3.20 +/- 0.27) microM in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.01). The content of SOD was (49.91 +/- 5.77) U/ml in VAPS 6.25 microg/ml group, (54.05 +/- 5.27) U/ml in the VAPS 12.5 microg/ml group, (57.44 +/- 5.70) U/ml in the VAPS 25 microg/mL group, as compared with model group there was statistically significant (P < 0.05). The content of GSH-Px was (172.50 +/- 18.65) U in VAPS 6.25 microg/ml group, (202.10 +/- 21.60) U in the VAPS 12.5 microg/ml group, (315.80 +/- 10.50) U in the VAPS 25 microg/ml group, the VAPS 12.5 microg/mL group and VAPS 25 microg/ml group was compared with model group, there were statistically significant difference (P < 0.01).
The VAPS have antioxidative damage effect of osteoarthritis cartilage cells within a certain range and dose-dependent manner. It may be the main mechanism for velvet antler polypeptides to treat osteoarthritis.