Jieduquyuziyin Prescription Suppresses the Inflammatory Activity of Macrophages via NOTCH1/NF-κB Pathway

Abstract

Jieduquyuziyin prescription (JP) is a traditional Chinese medicine (TCM) formula, which has been applied to the treatment of systemic lupus erythematosus (SLE) for decades, and its efficacy and safety have been confirmed in clinical practice. However, little is known about its molecular mechanism. To explore the effects of JP on macrophages' inflammatory activity and NOTCH1/NF-κB pathway. The JP-treated serum was prepared to determine its optimal concentration. Given the fact that active components in rats' serum might affect the results, control serum without JP components was prepared simultaneously. Lipopolysaccharide (LPS) was used to activate RAW264.7, and the cells were interfered with DAPT (NOTCH1 blocker), control serum, and JP-treated serum, respectively. After the above intervention, the expression of NOTCH1 and RBPJ, the nuclear translocation of NF-κB, and the extracellular release of IL6, TNFα, and NO, was evaluated by real-time reverse transcription-polymerase chain reaction (RT-PCR), western blotting (WB), enzyme-linked immunosorbent assay (ELISA), and Griess method. Both DAPT and JP-treated serum could significantly suppress the expression of NOTCH1 and RBPJ induced by LPS, as well as the nuclear translocation of NF-κB, leading to the decreased release of IL6, TNFα, and NO, while control serum had little effect on macrophage activity and NOTCH1/NF-κB pathway. These results demonstrated the effects of JP on macrophage activation and pro-inflammatory response and suggested that the molecular mechanism of JP might attribute to the inhibition of the NOTCH1/NF-κB pathway. Besides, previous studies suggested that paeoniflorin and ferulic acid are two major effective components in JP. In subsequent experiments, we would further explore the effects of these two components on MRL/lpr mice and macrophage activity.