Overexpression of HMBOX1 mediates NF-κB/CCL2 signaling pathway to inhibit the infiltration and activation of pulmonary macrophages induced by COPD
This study was designed to explore whether overexpression of HMBOX1 inhibits COPD-induced infiltration and pulmonary macrophage activation by regulating NF-κB/CCL2 signaling pathway.Forty Wistar rats were randomly divided into control group,chronic obstructive pulmonary disease group(COPD group),COPD+control overexpression group(COPD+ov-NC group)and chronic COPD+HMBOX1 overexpression group(COPD+ov-HMBOX1 group),with 10 rats in each group.The COPD model was established by continuous cigarette incense and intermittent intratracheal injection of lipopolysaccharide,wihle the HMBOX1 overexpression treatmet was carried out by intratracheal instillation of HMBOX1 overexpressing adenovirus.Western blot was used to detect the expression of HMBOX1,p-NF-κB and NF-κB proteins in lung tissue of rats;RT-qPCR was used to detect the mRNA expression of HMBOX1 and CCL2 in rat lung tissue;HE staining was used to observe the pathological changes of lung tissue in rats;ELISA was applied to detect the levels of TNF-α,MIP-2,IL-1β and IL-10 in serum and BALF of rats.Furthermore,the ratio of CD11b+F4/80+cells in lung tissue of rats was detected by immunofluorescence,while the ratio of F4/80+MHC Ⅱ+cells and F4/80+CD80+cells in lung tissue of rats was detected by flow cytometry.In control group,the alveolar structure of rats was intact,and no inflammatory cell infiltration was found.In COPD group and COPD+ov-NC group,a large number of inflammatory cells infiltrated into the lung tissue and alveolar structure was damaged.In COPD+ov-HMBOX1 group,there were fewer inflammatory cells infiltrated in lung tissue,and the damage of alveolar structure was alleviated.Compared with the control group,the mRNA and protein expression of HMBOX1 in lung tissue,the levels of IL-10 in serum and BALF,the levels of TNF-α,MIP-2 and IL-1β in serum and BALF,CD11b+F4/80+cells,F4/80+MHC Ⅱ+cells and F4/80+CD80+cells in lung tissue of rats in COPD and COPD+ov-NC groups were significantly decreased.HMBOX1 overexpression could revers the changes mentioned above in the two COPD groups.Taken together,overexpression of HMBOX1 can alleviate COPD-induced airway inflammation and lung injury,and its mechanism may be related to inhibiting infiltration and abnormal activation of macrophages in lung tissue mediated by activation of NF-κB/CCL2 signaling pathway.
COPDHMBOX1Macrophage infiltrationMacrophage activationNF-κB/CCL2 signal pathway
万方数据
维普
