Effect of TNF-α and Yigu capsule drug-containing sera on osteoblasts apoptosis in coculture system

AIM:To observe the effect of TNF-α and Yigu capsule drug-containing sera on osteoblasts apoptosis in osteoblasts-osteoclasts coculture system.METHODS:(1)Twenty female Sprague-Dawley rats,10 months old,were randomly assigned into 2 groups,NS and Yigu capsule groups,to prepare blank sera and drug-containing sera.(2)The DNA gel electrophoresis method was used to detect apoptosis of osteoblasts treated with different concentration of TNF-α in order to determine the best dosage in the co-culture system.(3)The cells were divided into four groups,TNF-α group,normal group,TNF-α + blank serum group and TNF-α + drug-containing serum group.DNA gel electrophoresis,acridine orange staining and flow cytometry were used to observe osteoblast apoptosis in these groups.RESULTS:(1)After induced by TNF-α for 72 h at a concentration of 60 μg/L,relatively typical DNA ladder appeared in TNF-α group.(2)Only two DNA brands appeared and most of cells were well-proportioned stained in TNF-α + drug-containing serum group,the rate of osteoblasts apoptosis in TNF-α + drug-containing serum group(9.60%±0.26%)was obviously lower than that in the TNF-α group(26.90%±0.06%)and TNF-α + blank serum group(18.10%±0.06%).CONCLUSION:TNF-α induces osteoblast apoptosis in the co-culture system,and Yigu capsule drug-containing serum prevents osteoblast apoptosis induced by TNF-α.