Effects of Rhizoma Dioscoreae Nipponicae total saponin -medicated serum on NF-κB p65 activity, STAT3 protein expression,and VEGF mRNA expression in IL-17- and TNF-α-induced rat synovial cell strain RSC-364

To evaluate the effects of Rhizoma Dioscreae Nipponicae(RDN) total saponin on angiogenesis of rheumatoid arthritis,we observed the effects of RDN total saponin-medicated serum on NF-κB p65 activity,STAT3 protein expression and VEGF mRNA expression in IL-17 and TNF-α-induced rat synovial cell strain RSC-364.Medicated serum of total saponin from RDN and Tripterygium(positive control group) were prepared.Then rat synovial cell strain RSC-364 was stimulated by IL-17(10 μg/L) + TNF-α(10 μg/L) in the presence or absence of RDN total saponin-medicated serum or Tripterygium-medicated serum.After twenty-four hours incubation,the NF-κB p65 DNA-binding activity in nuclear extract of rat synovial cell strain RSC-364 was detected by TransAM TM NF-κB p65 Transcription Factor Assay Kit.Western blot was used to observe the expression of STAT3 protein in each cells group,while VEGF mRNA expression in rat synovial cell strain RSC-364 in each group was detected by PrimeScript TM real-time quantitative PCR detection kit.Compared with the control group,the NF-κB p65 DNA-binding activity and the expressions of STAT3 protein and VEGF mRNA were all increased remarkable in rat synovial cell strain RSC-364 induced by IL-17+TNF-α model group(P0.01,P0.01,P0.05).While the NF-κB p65 DNA-binding activity and the expressions of STAT3 protein and VEGF mRNA in Tripterygium medicated serum group and RDN total saponin-medicated serum group were remarkably lower than those of IL-17+ TNF-α model group(P0.01,P0.01,P0.05).There was no significant difference between the two medicated serum groups(P0.05).In conclusion,RDN total saponin-medicated serum could remarkably lower NF-κB p65 DNA-binding activity and STAT3 expression,thus regulate VEGF secretion and restrain angiogenesis of rheumatoid arthritis.