Effect of Bugu granules in a drug-containing serum on chondrocyte apoptosis and the Trx2 signaling pathway.

Traditional Chinese medicine for invigorating the kidney and promoting blood circulation is commonly prescribed for the treatment of osteoarthritis associated with kidney deficiency and blood stasis. However, the specific mechanisms of these medicines are still unclear. The present study aimed to evaluate the protective effects of Bugu granules against sodium nitroprusside-induced chondrocyte apoptosis and elucidate the underlying molecular mechanisms. Drug-containing serum was prepared by administering rats with Bugu granules and harvesting the serum. Chondrocytes were exposed to different dilutions of serum, and apoptosis assessed by flow cytometry after staining with annexin V­FITC/PI. Flow cytometry showed that chondrocyte apoptosis increased significantly after incubation with 2 mol/L sodium nitroprusside for 24 h (t = -48.221, P = 0.000), and the apoptotic rate of chondrocytes decreased with increasing concentrations of drug-containing serum (F = 33.965, P = 0.000). Cellular levels of Trx2, ASK1, caspase­3, and reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay. The cellular content of Trx2 increased gradually with increasing concentrations of drug-containing serum (F = 2610.593, P = 0.000), while that of ASK1 (F = 2473.545, P = 0.000), caspase­3 (F = 209.921, P = 0.000), and ROS (F = 1666.435, P = 0.000) all decreased significantly. The mRNA expression levels were analyzed by RT-qPCR, which revealed that expression levels of Trx2 and caspase­3 mRNA increased and decreased significantly, respectively, following exposure to Bugu granules in the drug-containing serum (F = 6.974, P = 0.003 and F = 3.691, P = 0.191; respectively), but the expression of ASK1 mRNA was not significantly different between treatment groups (F = 1.784, P = 0.191). Taken together, these results support the hypothesis that the Trx2 signaling pathway is activated by Bugu granules, which in turn inhibits chondrocyte apoptosis. This may play a role in preventing the development of osteoarthritis.