Doxycycline alleviates paraquat-induced acute lung injury by inhibiting neutrophil-derived matrix metalloproteinase 9

Paraquat (PQ), a highly toxic herbicide, selectively accumulates in the lungs and causes pulmonary damage through oxidative and inflammatory processes after intentional or accidental poisoning. The resulting acute lung injury (ALI) is characterized by neutrophil infiltration and extensive inflammation with rapid respiratory failure. However, effective therapies are lacking. We tested the hypothesis that suppressing neutrophil-derived matrix metalloproteinase 9 (MMP9) would ameliorate the inflammatory milieu and alleviate PQ-induced ALI. Lung injury was assessed in mice intratracheally injected with PQ aerosol by measuring the lung static compliance, cell count and neutrophil percentage of the bronchoalveolar lavage fluid (BALF) and lung, alveolar-capillary permeability, and histopathological lung injury scores. MMP9/2 activity was assessed by gelatin zymography, and the location of neutrophils and MMP9 in the lung was evaluated by immunofluorescence costaining. In the neutrophil depletion experiment, mice received anti-Ly6G antibody intraperitoneally; for the MMP inhibition experiment, an MMP inhibitor, doxycycline (DOX), was administered by gavage. In PQ-induced ALI, the activity of neutrophil-derived MMP9 but not MMP2 increased significantly. Neutrophil depletion reduced the inflammatory burden, improved pulmonary edema, and reduced the PQ-induced overexpression of MMP9. Consistently, oral delivery of DOX to mice decreased the overexpression of MMP9 that was activated by PQ and phenocopied the resolution of PQ-induced ALI observed after neutrophil depletion. Taken together, our results show for the first time that DOX is involved in the resolution of PQ-induced ALI via a mechanism involving reducing the activity of neutrophil-derived MMP9. We speculate that DOX may represent a novel therapeutic strategy for PQ-induced ALI.