Background： Dendritic cells are professional antigen-presenting cells found in an immature state in epithelia and interstitial space, where they capture antigens such as pathogens or damaged tissue. Matrix metallopep...Background： Dendritic cells are professional antigen-presenting cells found in an immature state in epithelia and interstitial space, where they capture antigens such as pathogens or damaged tissue. Matrix metallopeptidase 13 （MMP- 13）, a member of the collagenase subfamily, is involved in many different cellular processes and is expressed in murine bone marrow-derived dendritic cells （DCs）. The function of MMP-13 in DCs is not well understood. Here, we investigated the effect of MMP-13 on DC maturation, apoptosis, and phagocytosis. Methods： Bone marrow-derived dendritic cells were obtained fiom C57BL/6 mice. One short-interfering RNA specific for MMP- 13 was used to transfect DCs. MMP-13-silenced DCs and control DCs were prepared, and apoptosis was measured using real-time polymerase chain reaction and Western blotting. MMP-13-silenced DCs and control DCs were analyzed for surface expression of CD80 and CD86 and phagocytosis capability using flow cytometry. Results： Compared to the control DCs, MMP- 13-silenced DCs increased expression of anti-apoptosis-related genes, BAGl （control group vs. MMP-13-silenced group： 4.08 ± 0.60 vs. 6.11 ± 0.87, P = 0.008）, BCL-2 （control group vs. MMP-13-silenced group： 7.54 ± 0.76 vs. 9.54 ± 1.29, P = 0.036）, and TP73 （control group vs. MMP- 13-silenced group： 4.33 ± 0.29 vs. 5.60 ±0.32, P = 0.001 ） and decreased apoptosis-related genes, CASPI （control group vs. MMP- 13-silenced group： 3.79±0.67 vs. 2.54±0.39, P - 0.019）, LTBR （control group vs. MMP- 13-silenced group： 9.23 ±1.25 vs. 6.24 ± 1.15, P = 0.012）, and CASP4 （control group vs. MMP-13-silenced group： 2.07 ± 0.56 vs. 0.35 ±0.35, P = 0.002）. Protein levels confirmed the same expression pattern. MMP- 13-silenced groups decreased expression of CD86 on DCs; however, there was no statistical difference in CD80 surface expression. Furthermore, MMP-13-silenced groups exhibited weaker phagocytosis capability. Conclusion： These results indicate that MMP-13 inhibition dampens DC maturation, apoptosis, and phagocytosis.展开更多
文摘Background： Dendritic cells are professional antigen-presenting cells found in an immature state in epithelia and interstitial space, where they capture antigens such as pathogens or damaged tissue. Matrix metallopeptidase 13 （MMP- 13）, a member of the collagenase subfamily, is involved in many different cellular processes and is expressed in murine bone marrow-derived dendritic cells （DCs）. The function of MMP-13 in DCs is not well understood. Here, we investigated the effect of MMP-13 on DC maturation, apoptosis, and phagocytosis. Methods： Bone marrow-derived dendritic cells were obtained fiom C57BL/6 mice. One short-interfering RNA specific for MMP- 13 was used to transfect DCs. MMP-13-silenced DCs and control DCs were prepared, and apoptosis was measured using real-time polymerase chain reaction and Western blotting. MMP-13-silenced DCs and control DCs were analyzed for surface expression of CD80 and CD86 and phagocytosis capability using flow cytometry. Results： Compared to the control DCs, MMP- 13-silenced DCs increased expression of anti-apoptosis-related genes, BAGl （control group vs. MMP-13-silenced group： 4.08 ± 0.60 vs. 6.11 ± 0.87, P = 0.008）, BCL-2 （control group vs. MMP-13-silenced group： 7.54 ± 0.76 vs. 9.54 ± 1.29, P = 0.036）, and TP73 （control group vs. MMP- 13-silenced group： 4.33 ± 0.29 vs. 5.60 ±0.32, P = 0.001 ） and decreased apoptosis-related genes, CASPI （control group vs. MMP- 13-silenced group： 3.79±0.67 vs. 2.54±0.39, P - 0.019）, LTBR （control group vs. MMP- 13-silenced group： 9.23 ±1.25 vs. 6.24 ± 1.15, P = 0.012）, and CASP4 （control group vs. MMP-13-silenced group： 2.07 ± 0.56 vs. 0.35 ±0.35, P = 0.002）. Protein levels confirmed the same expression pattern. MMP- 13-silenced groups decreased expression of CD86 on DCs; however, there was no statistical difference in CD80 surface expression. Furthermore, MMP-13-silenced groups exhibited weaker phagocytosis capability. Conclusion： These results indicate that MMP-13 inhibition dampens DC maturation, apoptosis, and phagocytosis.
