基于RAW264.7细胞炎症模型探讨秦皮乙素抗炎及潜在分子机制OA

Objective:To evaluate the anti-inflammatory activity of Esc in LPS-induced RAW264.7 cells and to investigate the underlying mechanisms involving STAT3 and S1PR1.Methods:RAW264.7 cells were pretreated with 1 μmol Esc for 20 h and then stimulated with 1 μg/ml LPS for 4 h to establish an inflammatory model.qRT-PCR,ELISA,and Western blot were used to detect the mRNA expression and secretion of TNF-α and IL-6,as well as the protein levels of p-STAT3 and S1PR1.Co-IP and IF were performed to assess STAT3 dimerization and p-STAT3 nu-clear translocation.Overexpression and siRNA experiments were used to explore the regulatory relationship between STAT3 and S1PR1 and their effects on inflammatory factors.Results:Esc significantly inhibited LPS-induced TNF-α and IL-6 mRNA expression and secretion,and decreased p-STAT3 protein levels without significantly changing S1PR1 protein abundance.Co-IP and IF showed that Esc suppressed STAT3 dimerization and p-STAT3 nuclear translocation.STAT3 and S1PR1 reciprocally regulated each other at the mRNA level,and overexpression of STAT3 or S1PR1 increased pro-inflammatory cytokine levels.Si-STAT3 or Si-S1PR1 markedly attenuated or abolished the inhibitory effects of Esc on TNF-α and IL-6.Conclusion:Esc effectively suppresses the LPS-induced inflammatory response in RAW264.7 cells,and its effect may be mainly associated with the inhibition of STAT 3 activation.Meanwhile,the influ-ence of Esc on S1PR1 may primarily involve functional regulation rather than changes in S1PR1 protein expression levels.