| Objective:To investigate effects of Fuyuan capsule(FC) on synthesis of collagen type Ⅱ(COL2A1) and aggrecan(ACAN) by regulating the TGF-β signaling pathway in osteoarthritis(OA)chondrocytes.Methods:Interleukin 1 beta( IL-1beta) induced Human chondroblastoma cells( SW1353) were treated by serum containing FC of different concentrations. Then the optimum concentration and time of serum containing FC were choosed and used in a follow-up experiment. SW1353 were cultured in vitro and were randomly divided into blank control group, IL-1β group,combination group 1(treated with IL-1β and serum containing FC), combination group 2( treated with IL-1β and SB431542) and combination group 3( treated with IL-1β、SB431542 and serum containing FC).The optimal intervention concentration of serum containing FC was set at 15 % and optimal intervention time was set at 72 h. Cell growth activities were measured by CCK8 assays 。 Cell cycle and apoptosis rate were analyzed by flow cytometry. The COL2A1 and ACAN m RNA expression were examined by quantitative reverse transcription polymerase chain reaction(q RT-PCR). The protein leve of phosphorylated smad 3 was tested by Western blot.Results:The optimum concentration and time of serum containing FC on cell proliferation was 15 percent serum containing FC at 72 h, while inducing S-phase cell cycle arrest, enhanced cell proliferation and protected the cells against IL-1β- and/or SB431542-induced cell growth inhibition(P < 0.05). Furthermore, FC reversed the decreased expression of COL2A1 and ACAN induced by IL-1β and SB431542, and blocked the decreased phosphorylation of Smad 3 induced by IL-1β alone or in combination with SB431542(P < 0.05).Conclusion:Our results suggest that FC promotes COL2A1 and ACAN expression as well as Smad3 activation by activating TGF-β signal pathway but incompletely depending on it in IL-1β-induced chondrocytes, thus protecting the cartilage from injury. |
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