Effects of medicated serum containingDuhuojisheng Decoction on the expression of cytochrome C, proCaspase-9 and proCaspase-3 in degenerative chondrocytes

Wang Wu-lian; Lin Yu; Zhang Yi-yuan; Xiao Li-li; Feng Er-you

doi:10.3969/j.issn.2095-4344.2015.33.005

Abstract：BACKGROUND:Previous studies have shown that apoptosis, a central feature of articular chondrocytes, plays a dominant role in cartilage damage, which is one of the pathological factors of articular cartilage degeneration. OBJECTIVE:To observe the effects of meditcated serum containingDuhuojisheng Decoction on the expression of cytochrome C, proCaspase-9 and proCaspase-3 in rat degenerative chondrocytes in vitro and to investigate the possible molecular biological mechanism ofDuhuojisheng Decoction in the treatment of knee osteoarthritis. METHODS:A cultivation system of degenerative chondrocytes in vitro was established. After treatment with meditcated serum containingDuhuojisheng Decoction or blank serum for 24 and 48 hours, the protein expression of cytochrome C, proCaspase-9 and proCaspase-3 was measured by western blot assay. RESULTS AND CONCLUSION: In the cytoplasm, the release of cytochrome C was reduced gradualy in both groups in a time-dependent manner, and the release amount of cytochrome C was significantly lower in the medicated serum group than the blank serum group (P < 0.05). In mitochondria, cytochrome C leakage was gradualy decreased in both groups, and it was decreased significantly in the medicated serum group compared with the blank serum group (P < 0.05). The protein expression of proCaspase-9 and proCaspase-3 was gradualy increased in both groups, especialy in the medicated serum group; the medicated serum containingDuhuojisheng Decoction could promote the protein expression of proCaspase-9 and proCaspase-3 in a time-dependent manner, and there was a significant difference at 24 and 48 hours (P< 0.01). These findings indicate that the medicated serum containingDuhuojisheng Decoction can inhibit the apoptosis of osteoarthritis chondrocytes through inhibiting the release of cytochrome C and the activation of Caspase-9 and Caspase-3.