Dissection of the role of paeoniflorin in the traditional Chinese medicinal formula Si-Ni-San against contact dermatitis in mice

Aims: The roles of specific active ingredients in Chinese medicinal formulas have not been clearly elucidated. In this Study, we selectively deleted and replenished paeoniflorin from Si-Ni-San, a traditional Chinese prescription, and aimed to identify the molecular basis of how paeoniflorin exerted its effect in Si-Ni-San. Main methods: Contact dermatitis was induced in mice with picryl chloride. Paeoniflorin was selectively deleted from Si-Ni-San by an immunoaffinity column. Quantitative real-time PCR, western blot, and enzyme-linked immunosorbent assay were used in this study. Key findings: Both Si-Ni-San and paeoniflorin significantly reduced ear swelling in mice while the paeoniflorin-deleted Si-Ni-San (Si-Ni-San(PF-)) showed little ameliorative effect. In lipopolysaccharide. evoked macrophages, Si-Ni-San and paeoniflorin markedly inhibited tumor necrosis factor-alpha production, cyclooxygenase-2 activity, as well as extracellular signal-regulated kinase 1/2 phosphorylation while Si-Ni-San(PF-) exhibited no or slight inhibitory effect. Furthermore, the inhibitory effect on the production of tumor necrosis factor-alpha reappeared when different proportions of paeoniflorin were replenished in Si-Ni-San(PF-). In addition, the expression of macrophage migration inhibitory factor in T cells, rather than macrophages, was significantly inhibited by Si-Ni-San, but not Si-Ni-San(PF-). Our data indicate paeoniflorin is the principal component of Si-Ni-San, exerting negative regulation on the function of macrophages in contact dermatitis. Significance: The present study suggests that dissecting the role of specific constituents in medicinal formulas through selective deletion and replenishment may be a useful strategy in recognizing and validating an active ingredient in traditional Chinese medicine. (C) 2009 Elsevier Inc. All rights reserved.