ABSTRACT
Emodin, an anthraquinones component of Rheum palmatun, has been used for anti-inflammatory purposes. However, its underlying molecular effect(s) on target cells remain to be well clarified. our current study was aimed at investigating the regulatory effect of Emodin on lipopolysaccharide induced inflammatory responses in RAW 264.7 macrophages by gelatin zymography . It was found that treatment of LPS(5μg/ml) for 18 h elevates the expression of MMP-9 and MMP-2 proteins in RAW 264.7 cells. It was also found that co-treatment of RAW 264.7 with drug Emodin(6μg/ml) and LPS(5μg/ml) for the period of 18 h reduces the expression level of MMP-9 and MMP-2 protein. Our data suggest that Emodin plays roles in regulating the digestion of Extracellular matrix by down-regulating the expression of MMP-9 and MMP-2 proteins.
ABSTRACT
Objective:To study the effect of IL-24 on the differentiation and function of osteoclasts.Methods:Mature osteoclasts were isolated from long bones of neonate rats.Optimal multiplicity of infection(MOI)of AdCMV-EGFP was determined.Then osteo-clasts and RAW264.7 cells were transfected with AdCMV-EGFP and AdCMV-IL-24 respectively.The function of osteoblasts was studied by the observation of bone resorption lacunae,the differentiation of RAW264.7 cells was evaluated by the expression of osteo-clast related genes examined with real-time PCR.Results:Osteoclasts were TRAP positive with more than 2 neclei.MOI=400 was suitable for AdCMV-EGFP transfection.The resorption lacunae area in AdCMV-IL24 transfected cells was larger than that in AdCMV-EGFP transfected cells(P<0.05).Real-time PCR showed that under induced conditions,osteoclastic related genes NFATc1,CTSK and TRAP were changed in RAW264.7 cells transfected with AdCMV-IL-24.Conclusion:IL-24 may promote the differentiation and function of osteoclasts.
ABSTRACT
Objective:To establish an experimental model for intracellular antibacteria and endotoxin neutralization in vitro to detect the antibacterial and endotoxin neutralization activity of the muBPI_(25) protein.Methods: RAW264.7 cells were transfected with pcDNA3.1(+)muBPI_(36-259), and then were infected with intracellular bacterial of either G ~+/G~-to establish the experimental model of intracellalar antibacteria.The RAW264.7 cells were co-transfected with the pSecTag2B-muBPI_(36-259) and dual-luciferase reporter gene plasmids for establishment of the experimental model of endotoxin neutralization.Results:The experimental model of intracellular antibacteria confirmed that the muBPI_(25) protein could inhibit/kill Salmonella typhi.The experimental model of endotoxin neutralization indicated that the muBPI_(25) protein could neutralize endotoxin.Conelusion: We firstly demonstrate that murine BPI N-terminal functional fragment(muBPI_(25) protein)can inhibit/kill Salmonella typhi,and can neutralize, its lysating product, endotoxin.