An increasing number of studies have focused on the phenomenon that
mitochondrial DNA (
mtDNA) activates innate immunity responses. However, the specific role of
mtDNA in inflammatory
lung disease remains elusive. This study was designed to examine the proinflammatory effects of
mtDNA in lungs and to investigate the putative mechanisms. C57BL/6 mice were challenged intratracheally with
mtDNA with or without pretreatment with
chloroquine. Changes in pulmonary histopathology,
cytokine concentrations, and phosphorylation levels of
p38 MAPK were assayed at four time points. In in vitro experiments, THP-1 macrophages were pretreated or not pretreated with
chloroquine, TLR9
siRNA,
p38 MAPK siRNA, or
SB203580 and then incubated with
mtDNA. The levels of
cytokines and p-p38 MAPK were detected by ELISA and Western blot, respectively. The intratracheal administration of
mtDNA induced infiltration of inflammatory cells, production of proinflammatory
cytokines (including IL-1β, IL-6, and TNF-α), and activation of
p38 MAPK. The
chloroquine pretreatment resulted in an abatement of
mtDNA-induced local
lung inflammation. In vitro experiments showed that the exposure of THP-1 macrophages to
mtDNA also led to a significant upregulation of IL-1β,
IL-6, and TNF-α and the activation of
p38 MAPK. And these responses were inhibited either by
chloroquine and TLR9
siRNA or by
SB203580 and
p38 MAPK siRNA pretreatment. The intratracheal administration of
mtDNA induced a local inflammatory response in the mouse lung that depended on the interactions of
mtDNA with TLR9 and may be correlated with infiltrating macrophages that could be activated by
mtDNA exposure via the TLR9-p38 MAPK signal transduction pathway.