Cigarette smoking is the primary risk factor for
COPD which is characterized by excessive
inflammation and airflow obstruction of the lung. While
inflammation is causally related to initiation and progression of
COPD, the mitochondrial mechanisms that underlie the associated inflammatory responses are poorly understood. In this context, we have studied the role played by
Mitogen activated
protein (
MAP) kinase kinase 3 (MKK3), a dual-specificity
protein kinase, in cigarette
smoke induced-
inflammation and
mitochondrial dysfunction. Serum pro-inflammatory
cytokines were significantly elevated in WT but not in MKK3-/- mice exposed to Cigarette
smoke (CS) for 2 months. To study the cellular mechanisms of
inflammation, bone marrow derived macrophages (BMDMs), wild type (WT) and MKK3-/-, were exposed to cigarette
smoke extract (CSE) and inflammatory
cytokine production and mitochondrial function assessed. The levels of IL-1β,
IL-6, and TNFα were increased along with higher
reactive oxygen species (ROS) and P-NFκB after CSE treatment in WT but not in MKK3-/- BMDMs. CSE treatment adversely affected basal mitochondrial respiration,
ATP production, maximum respiratory capacity, and spare respiratory capacity in WT BMDMs only. Mitophagy, clearance of dysfunctional mitochondria, was up regulated in CS exposed WT mice lung tissue and CSE exposed WT BMDMs, respectively. The proteomic analysis of BMDMs by iTRAQ (isobaric tags for relative and absolute quantitation) showed up regulation of
mitochondrial dysfunction associated
proteins in WT and higher OXPHOS (Oxidative phosphorylation) and
IL-10 signaling
proteins in MKK3-/- BMDMs after CSE exposure, confirming the critical role of mitochondrial homeostasis. Interestingly, we found increased levels of p-MKK3 by immunohistochemistry in
COPD patient lung tissues that could be responsible for insufficient mitophagy and
disease progression. This study identifies MKK3 as a negative regulator of mitochondrial function and inflammatory responses to CS and suggests that MKK3 could be a therapeutic target.