ABSTRACT
Objective To investigate the effect of Dectin-1 on the release of inflammatory factors interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in rat tracheal epithelial cells (RTECs) stimulated with heat-treated Candida glabrata (C.glabrata).Methods RTECs cultivated in vitro were randomly divided into three groups,including control group(RTECs + sterile normal saline),fungal stimulation group(RTECs + heat-treated C.glabrata),and inhibitor intervention group(RTECs + laminarin + heat-treated C.glabrata),cells were harvested after incubation for 0,2,4,6 hours respectively,cell survival rate was determined by MTT method,expression of Dectin-1 was analyzed by Western Blot,expression of IL-10 and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA).Results Heat-treated C.glabrata destroyed cell structure and reduced cell survival rate.At the beginning of the culture (0 h),cell survival rate,expressions of Dectin-1,IL-10 and TNF-α among three groups were all not significantly different(all P＞0.05).After incubation for 2,4,6 hours,expressions of Dectin-1,IL-10 and TNF-α in fungal stimulation group and inhibitor intervention group were both significantly higher than control group;expressions of Dectin-1,IL-10,and TNF-α in inhibitor intervention group was lower than fungal stimulation group(all P＜0.05).The expressions of IL-10 in inhibitor intervention group at 0 h and 2 h was not significantly different,expressions of Dectin-1,IL-10,and TNF-α in fungal stimulation group and inhibitor intervention group at different incubation periods were significantly different(all P＜0.05).Conclusion Dectin-1 is an important receptor for RTECs to recognize the heat-treated C.glabrata,it induces the release of IL-10 and TNF-α,and mediate the occurrence of inflammation.
ABSTRACT
Objective To explore the effects of co‐cultured heat‐treated candida glabrata with rat tracheal epithelial (RTE) cells on the expression of Dectin‐1 and the production of IL‐6 and TNF‐α.Methods RTE cells in vitro were co‐cultured with heat‐treated candida glabrata bacteria liquid for 2 ,4 ,6 h ,while without co‐cultured RTE cells were used as control group .We observed the morphological changes of RTE cells ,detected the protein expressions of Dectin‐1 by Western blot ,used real‐time PCR to detecte the mRNA expressions of IL‐6 and TNF‐αand measured protein expression of IL‐6 and TNF‐αby ELISA .Results With the pass‐ing of time ,the RTE cells were damaged extensively and the expression of Dectin‐1 ,IL‐6 and TNF‐αbecame more and more signifi‐cant .Obviously ,there had significant difference in the expression of Dectin‐1 ,IL‐6 and TNF‐α between the co‐cultured 2 h group and the control group ,the co‐cultured 4 h group and the co‐cultured 2 h group ,the co‐cultured 6 h group and the co‐cultured 4 h group (P<0 .05) .Conclusion RTE cells have natural immune function .The Dectin‐1 involves in the recognition of heat‐treated Candida glabrata ,activating secretion of IL‐6 and TNF‐αand mediating inflammatory reaction .IL‐6 plays a negative regulation role .