BACKGROUND:For knee osteoarthritis,synovium inflammation and abnormal proliferation and activation of fibroblast-like synoviocytes caused by insuffcient apoptosis are important driving factors for disease progression.Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway plays a key role in regulating inflammatory response and apoptosis of synovial cells.As a traditional Mongolian medicine component,Agiophyllum Oligo Saccharides has anti-inflammatory and immunomodulatory functions,but its specific molecular mechanism has not been fully revealed.OBJECTIVE:To explore the effects and mechanism of Agiophyllum Oligo Saccharides on lipopolysaccharide-induced inflammation and apoptosis of mouse synovial cells.METHODS:The mouse synovial cells were randomly divided into a control group,a model group,an Agiophyllum Oligo Saccharides group and a Bay 11-7082 group.Osteoarthritic synoviocyte model was prepared using 1.0 μg/mL lipopolysaccharide intervention for 36 hour.The Agiophyllum Oligo Saccharides group was treated with 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours;and the Bay 11-7082 group was firstly incubated for 4 hours with 1 μmol/L NF-κB pathway inhibitor Bay 11-7082,and then 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours.Western blot was used to detect TLR4,MyD88,NF-κBp65,p-NF-κBp65,and IκBα protein expression.The level of tumor necrosis factor-α and interleukin-1β in the supernatant were analyzed by ELISA.To detect the expressions of Bax and Bcl-2,western blot and immunofluorescence analysis were performed.The intracellular reactive oxygen species level was detected by DCFH-DA.RESULTS AND CONCLUSION:(1)Compared with the control group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 increased(P<0.05),whereas the protein expressions of IκBα and Bax decreased(P<0.05)in the model group.(2)Compared with the model group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 decreased(P<0.05),while the expressions of IκBα and BAX increased(P<0.05)in the Agiophyllum Oligo Saccharides group.(3)The levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species and the protein expression of Bcl-2 in the Bay 11-7082 group were higher than those in the Agiophyllum Oligo Saccharides group(P<0.05),but still lower than those in the model group(P<0.05),whereas the expression of Bax protein was lower than that in the Agiophyllum Oligo Saccharides group(P<0.05),but still higher than that in the model group(P<0.05).(4)Compared with the model group and the Agiophyllum Oligo Saccharides group,the protein expressions of TLR4,MyD88 and p-NF-κBp65/NF-κBp65 decreased(P<0.05),whereas the protein expressions of IκBα increased(P<0.05)in the Bay 11-7082 group.To conclude,Agiophyllum Oligo Saccharides could inhibit inflammation of mouse synovial cells and promote apoptosis of synovial cells with inflammation,and the mechanisms may be correlated with regulating the TLR4/NF-κB signaling pathway.

BACKGROUND:For knee osteoarthritis,synovium inflammation and abnormal proliferation and activation of fibroblast-like synoviocytes caused by insuffcient apoptosis are important driving factors for disease progression.Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway plays a key role in regulating inflammatory response and apoptosis of synovial cells.As a traditional Mongolian medicine component,Agiophyllum Oligo Saccharides has anti-inflammatory and immunomodulatory functions,but its specific molecular mechanism has not been fully revealed.OBJECTIVE:To explore the effects and mechanism of Agiophyllum Oligo Saccharides on lipopolysaccharide-induced inflammation and apoptosis of mouse synovial cells.METHODS:The mouse synovial cells were randomly divided into a control group,a model group,an Agiophyllum Oligo Saccharides group and a Bay 11-7082 group.Osteoarthritic synoviocyte model was prepared using 1.0 μg/mL lipopolysaccharide intervention for 36 hour.The Agiophyllum Oligo Saccharides group was treated with 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours;and the Bay 11-7082 group was firstly incubated for 4 hours with 1 μmol/L NF-κB pathway inhibitor Bay 11-7082,and then 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours.Western blot was used to detect TLR4,MyD88,NF-κBp65,p-NF-κBp65,and IκBα protein expression.The level of tumor necrosis factor-α and interleukin-1β in the supernatant were analyzed by ELISA.To detect the expressions of Bax and Bcl-2,western blot and immunofluorescence analysis were performed.The intracellular reactive oxygen species level was detected by DCFH-DA.RESULTS AND CONCLUSION:(1)Compared with the control group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 increased(P<0.05),whereas the protein expressions of IκBα and Bax decreased(P<0.05)in the model group.(2)Compared with the model group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 decreased(P<0.05),while the expressions of IκBα and BAX increased(P<0.05)in the Agiophyllum Oligo Saccharides group.(3)The levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species and the protein expression of Bcl-2 in the Bay 11-7082 group were higher than those in the Agiophyllum Oligo Saccharides group(P<0.05),but still lower than those in the model group(P<0.05),whereas the expression of Bax protein was lower than that in the Agiophyllum Oligo Saccharides group(P<0.05),but still higher than that in the model group(P<0.05).(4)Compared with the model group and the Agiophyllum Oligo Saccharides group,the protein expressions of TLR4,MyD88 and p-NF-κBp65/NF-κBp65 decreased(P<0.05),whereas the protein expressions of IκBα increased(P<0.05)in the Bay 11-7082 group.To conclude,Agiophyllum Oligo Saccharides could inhibit inflammation of mouse synovial cells and promote apoptosis of synovial cells with inflammation,and the mechanisms may be correlated with regulating the TLR4/NF-κB signaling pathway.