Effect Of Jieduquyuziyin Prescription On Expression Of IRAK1 In Peritoneal Macrophages Of MRL/lpr Lupus Mice

Objective To investigate the effect of Jieduquyuziyin prescription on the abnormal activation of IRAK1 signaling pathway in peritoneal macrophages of MRL/lpr mice,and to investigate the effect of Jieduquyuziyin Recipe on IRAK1-NF-?B signaling pathway.It provides good theoretical support for the treatment of systemic lupus erythematosus.Methods The drug-containing serum was prepared,and the peritoneal macrophages of MRL/lpr lupus mice were isolated in vitro,and the activity of Jieduquyuziyin prescription on cells was detected by CCK8 method.The cells were subjected to gene silencing and gene overexpression of lentiviral transfection,and the cells were randomly divided into blank serum group,LPS group,LPS plus Chinese medicine serum group,Control shRNA group,IRAK1 shRNA group,IRAK1 shRNA plus Chinese medicine serum group,LV-Control group,LV-IRAK1 group and LV-IRAK1 plus Chinese medicine serum group.Immunofluorescence staining was used to detect the effect of Jieduquyuziyin recipe on the expression of IRAK1 in cells.The content of TNF-? and IL-6 in cell supernatant was determined by ELISA.The expression of IRAK1,NF-?B,TNF-? and IL-6 mRNA was detected by RT-qPCR.The expression levels of IRAK1,p-IRAK1,TRAF6,IKB?,p-IKB?,IKK?+IKK?,NF-?B and p-NF-?B were detected by Western-blot method.Results Jieduquyuziyin prescription serum contains 2.5% as the optimal concentration.LPS can up-regulate the expression of IRAK1 in cells by inducing activation of IRAK1-NF-?B signaling pathway,and increase TNF-? and IL-6 in cell supernatant.And also can increase expression of cellular IRAK1,NF-?B,TNF-? and IL-6 mRNA as well as IRAK1,p-IRAK1,TRAF6,IKB?,p-IKB?,IKK?+IKK?,NF-?B and p-NF-?B protein levels.The difference was statistically significant(P<0.05),and the serum of Jieduquyuziyin recipe could reduce these expression levels.In the interference of shRNA gene silencing and overexpression of lentivirus,shRNA can effectively inhibit the activation of IRAK1 signaling pathway and its downstream expression.Jieduquyuziyin prescription serum can further reduce the difference and the difference was statistically significant(P<0.05).Overexpression of lentivirus can activate the abnormal expression of IRAK1 signaling pathway,and significantly increase the expression level of IRAK1-NF-?B signaling pathway.Jieduquyuziyin drug-containing serum can reduce the expression level of IRAK1-NF-?B signaling pathway(P<0.05).Conclusion Jieduquyuziyin recipe can inhibit the secretion of cytokines TNF-? and IL-6,the expression of IRAK1,NF-?B,TNF-? and IL-6 mRNA as well as IRAK1,p-IRAK1,TRAF6,IKB?,p-IKB?,IKK?+IKK?,NF-?B and p-NF-?B protein levels.Jieduquyuziyin recipe may inhibit the activation of peritoneal macrophages in MRL/lpr mice by down-regulating the IRAK1-NF-?B signaling pathway,so IRAK1 may be a potential target for the treatment of SLE.