Effect of Jieduquyuziyin-prescription-treated Rat Serum on IRAK1 Signaling Pathway Expression of Mice Monocyte-macrophage Cells / 中国药学杂志

ABSTRACT

OBJECTIVE: To study the effect of serum containing Jieduquyuziyin-prescription (JP) on signal pathway of interleukin-1 receptor-associated kinase 1 (IRAK1) in mononuclear macrophages of mice stimulated by lipopolysaccharide (LPS), and to explore the effect of Jieduquyuziyin-prescription on IRAK1 and NF-κB inflammatory signaling pathways, which providing a good theoretical support for its anti-inflammatory clinical medication. METHODS: In this study, mice mononuclear macrophages cultured in vitro were randomly divided into blank group, LPS group, JP serum group, blank serum group, LPS plus JP serum group, LPS plus blank serum group, IRAK1 inhibitor group, inhibitor plus LPS group, inhibitor plus JP serum group and inhibitor plus blank serum group. After intervention for 24 h, the activity of JP on macrophages was tested by CCK8 method. The IRAK1 expression in macrophages was tested by immunofluorescence chemical staining. The content of TNF-α in the supernatant of the cells was detected by ELISA. The mRNA expressions of IRAK1, NF-κB, TNF-α and IL-6 were detected by RT-PCR. The protein expressions of IRAK1, p-IRAK1 and NF-κB were detected by Western-blot. The LC-MS was used to detect the active ingredients in JP serum. RESULTS: The results show that 2.5% of JP serum is the optimal concentration. Jieduquyuziyin-prescription could down-regulate the expression of TNF-α and IL-6 and inhibit the expression of IRAK1 and activate NF-κB(P<0.05). Paeoniflorin and ferulic acid were detected in the JP serum. CONCLUSION: Jieduquyuziyin-prescription can inhibit the expression of IRAK1 and NF-κB in mouse monocyte-macrophage cells after LPS stimulation and provide a good theoretical support for its anti-inflammatory clinical medication.