Background: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 con...Background: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.Objective:To explore the effects of JP on macrophages’inflammatory activity and NOTCH1/NF-κB pathway.Methods: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 simul-taneously.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 expres-sion 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.Results: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.Conclusion: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 path-way.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.展开更多
基金This work was financially supported by the Graduate Science Research Fund of Zhejiang Chinese Medical University(Grant No:2020YKJ03)the Young and Middle-aged Science Innovation Research Fund of Zhejiang Chinese Medical University(Grant No:KC201941).
文摘Background: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.Objective:To explore the effects of JP on macrophages’inflammatory activity and NOTCH1/NF-κB pathway.Methods: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 simul-taneously.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 expres-sion 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.Results: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.Conclusion: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 path-way.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.