Effects Of Oxidized Low-density Lipoprotein On The Expression Of CD47and MicroRNA-155of DC2.4Cells Of Mice

ObjectiveTo observe the influence of oxidized low-density lipoprotein (ox-LDL) on themorphology、phenotypic maturation and the expression of membrane protein CD47and microRNA-155of the murine dendritic cell line DC2.4cells，and investigate thepotential role of CD47and microRNA-155in the maturation and migration of DCunder the condition of atherosclerotic (AS)。Methods1、DC2.4were seeded in6-well plates, cultured for3days with RPMI1640completemedium which containing10%FBS,then in favor of non-serum medium and dividedinto2groups:the experimental group which treatment by ox-LDL-(100ug/mL), andthe control group by phosphate buffered saline (PBS).Culture changes in cellmorphology were observed under an inverted microscope; DC2.4cells were collectedafter12hours and24hours,and the expression of cell phenotype (MHC-II, CD80andCD86) were analyzed by flow cytometry.2、DC2.4were seeded in6-well plates, cultured for3days with RPMI1640completemedium which containing10%FBS,then in favor of non-serum medium and dividedinto2groups:the experimental group which treatment by ox-LDL(100ug/mL), andthe control group by phosphate buffered saline (PBS). DC2.4cells were collected after12hours and24hours, and the expression of CD47were analyzed by Westernblot method after the protein were extracted; and the expression of microRNA-155were analyzed by RT-PCR method fafter the RNA were extracted.Results1.1Observation on morphology of DC2.4: In control group, cells were round oroval, small, translucent, surrounding a small burr-like or spine-like protuberances;After12or24hours’ treatment by ox-LDL,the cells were observed an increasing insize,had gradual cytoplasm,in irregular multi-type state, with typical dendriticprocesses.1.2Phenotypic changes in DC2.4: Collected the suspension cells and detected themolecules expressed on the surface by Flow cytometry analysis.The results showed thatthe expression of MHC-II, CD80and CD86were compared between the three groups hadstatistically significant differences (P <0.05),ox-LDL groups higher than PBS group.2.1The expression of CD47were significant different between the three groups (P<0.05),ox-LDL groups lower than PBS group.2.2The expression of microRNA-155were significant different between the threegroups (P <0.05),ox-LDL groups higher than PBS group.Conclusions1.The phenotype expression of MHC-II, CD86and CD80in DC2.4after ox-LDLtreatment are significantly higer, ox-LDL can induce the mature of DC2.4.2. The phenotype expression of CD47are significantly inhibited and microRNA-155are significantly increased in DC2.4after ox-LDL treatment.