Effects Of CD38Gene Disruption On Migration And Release Of Inflammatory Factors In Macrophages Of Mouse

Objective：Inflammation plays an important role in the development of a variety of diseases,especially chronic disease, such as cardiovascular disease, cancer, diabetes and so on.There are many factors contributing to inflammation. Among all the influencingfactors, the activation of macrophages and the increasing expressions of variouscytokines such as MCP-1, TNF-α, IL-1β was considered to be much more important.CD38is a widespread multifunctional ectoenzyme that catalyzes the synthesis andhydrolysis of cyclic ADP-ribose (cADPR) from NAD+to ADP-ribose to adjust theconcentration of NAD+in mammals. Our previous experimental results showed thatthe disruption of CD38promoted atherosclerosis induced by high fat diet in mice andcould result in a significant invasion of macrophages in the plaque tissues. The aim ofthis proposal is to investigate the mechanism of CD38participating in the migrationand inflammatory response in macrophages through observing the cell migration andstudying the gene expression of inflammation related transcription factorFOXO1/NF-кB and their downstream target genes. Our data should provide aninsight for elucidating the exact function and molecular mechanism of CD38on theinflammation related signal pathway and a novel perspective of dealing with theinflammatory diseases.Method1. Primary culture of macrophages. The bone marrow-derived mononuclear cellswere collected and the macrophages were identified after cultured with7days andthen the macrophages would be used for follow-up experiments.2. The migration of macrophages. Wound-healing assay and macrophagemigration assay were performed in an effort to understand the migration of CD38-/-BMMs.3. Gene expression and inflammatory cytokine secretion of macrophages.FOXO1nuclear translocation was examined by immunofluorescence. Elisarray assay was performed in order to detect cytokine secretion of M1such as TNF-α, IL-1β andM2such as IL10, IFN, while the expression of MCP-1、TLR4、NF-κB were analyzedby Western Blotting.Results1. Mature macrophage-specifc marker F4/80expression was examined byimmunofluorescence. And F4/80-positive cells in WT and CD38-/-were higher90%for follow-up experiments.2. Defciency of CD38in BMMs resulted in increased migration.3. Defciency of CD38in BMMs resulted in increased FOXO1nucleartranslocation and MCP-1、TLR4、NF-κB expression. Cytokine secretion of M1suchas TNF-α, IL-1β were significantly increased, but there were no difference in M2cytokine secretion, such as IL10、IFN.4. Defciency of CD38in BMMs resulted in increased expression of Ccr2.ConclusionDefciency of CD38promoted migration and release of inflammatory factors inmacrophages. The activation of the FOXO1/CCR2pathway resulted in the increasedmigration of macrophages, whereas the TLR4/NF-κB pathway might be involved inthe increased cytokine secretions.