ABSTRACT
Introduction@#When human body encounters external pathogens primary/innate immunity cells are activated by recognizing them and secondary/adaptive immunity is activated consecutively. In our previous study, we revealed that there is a synergistic action between TLR9 and IFN-γ signaling in the endothelial cells. @*Purpose@#To determine the role of negative and positive regulator proteins on the IFN-γ/TLR9 signaling pathway. @*Methods@#In this study, murine endothelial cell (END-D) culture was used. END-D cells pre-treated with TLR9 ligand CpG DNA and then stimulated with IFN-γ. The negative (SHP-2, SOCS1, PIAS1) and positive (p38) regulator protein expression was detected by Western blotting. @*Results and Conclusion@#Treatment by TLR9 ligand CpG DNA and IFN-γ increased positive regulator p38 phosphorylation in 0.5 hour. CpG DNA inhibited IFN-γ negative regulator PIAS1 protein expression in 6 hour and SOCS1 and SHP-2 expression could not affect in 4 hour.
ABSTRACT
Objective To observe the effect of oxymatrine(OM) on the expressions of p-STAT1 and PIAS1 signaling molecules at protein and mRNA levels in the proliferation of the human mesangial cells(HMC) induced by lipopolysaccharide(LPS) and explore the relationship between them.Methods HMCs were primarily cultured from a 4-month-old aborted human fetus(with informed consent and approved by the Ethics Committee of Lanzhou University),and then divided into 3 groups,that is,control group,LPS group(10 ng/ml) and OM group(LPS 10 ng/ml and OM 320 mg/L).After cultured for 12,24 and 48 h respectively,HMC proliferation were analyzed by methyl thiazolyl tetrazolium(MTT) assay and type Ⅳ collagen in the supernatants were detected by ELISA.At the same time points,the cells lysates were collected for the mRNA and protein expressions of p-STAT1 and PIAS1 by real-time quantitative RT-PCR and Western blot analysis.Results The cell proliferation of LPS group was faster and the type Ⅳ collagen protein was increased more than the control group(P