Basic Investigation| Volume 355, ISSUE 5, P497-505, May 2018

Chronic Alcohol Ingestion Impairs Rat Alveolar Macrophage Phagocytosis via Disruption of RAGE Signaling



      Alcohol significantly impairs antioxidant defenses and innate immune function in the lung and increases matrix metalloproteinase 9 (MMP-9) activity. The receptor for advanced glycation end products (RAGE) is a well-characterized marker of lung injury that is cleaved by MMP-9 into soluble RAGE and has not yet been examined in the alcoholic lung. We hypothesized that chronic alcohol ingestion would impair RAGE signaling via MMP-9 in the alveolar macrophage and thereby impair innate immune function.

      Materials and Methods

      Primary alveolar macrophages were isolated from control-fed or alcohol-fed rats. Real-time polymerase chain reaction (qRT-PCR), Western blotting, and enzyme-linked immunosorbent assays were performed to evaluate RAGE expression. Silencing of MMP-9 ribonucleic acid (RNA) in a rat alveolar macrophage cell line was confirmed by qRT-PCR, and immunofluorescence (IF) was used to assess the association between alcohol, MMP-9, and RAGE. Phagocytosis was assessed using flow cytometry. Sulforaphane and glutathione were used to assess the relationship between oxidative stress and RAGE.


      RAGE messenger RNA expression was significantly increased in the alveolar macrophages of alcohol-fed rats, but IF showed that membrane-bound RAGE protein expression was decreased. Lavage fluid demonstrated increased levels of soluble RAGE (sRAGE). Decreasing MMP-9 expression using si-MMP-9 abrogated the effects of alcohol on RAGE protein. Phagocytic function was suppressed by direct RAGE inhibition, and the impairment was reversed by antioxidant treatment.


      Chronic alcohol ingestion reduces RAGE protein expression and increases the amount of sRAGE in alveolar lavage fluid, likely via cleavage by MMP-9. In addition, it impairs phagocytic function. Antioxidants restore membrane-bound RAGE and phagocytic function.

      Key Indexing Terms

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to The American Journal of the Medical Sciences
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Mehta A.J.
        • Guidot D.M.
        Alcohol abuse, the alveolar macrophage and pneumonia.
        Am J Med Sci. 2012; 343: 244-247
        • Moss M.
        • Steinberg K.P.
        • Guidot D.M.
        • et al.
        The effect of chronic alcohol abuse on the incidence of ARDS and the severity of the multiple organ dysfunction syndrome in adults with septic shock: an interim and multivariate analysis.
        Chest. 1999; 116: 97S-98SS
        • Happel K.I.
        • Nelson S.
        Alcohol, immunosuppression, and the lung.
        Proc Am Thorac Soc. 2005; 2: 428-432
        • Joshi P.C.
        • Guidot D.M.
        The alcoholic lung: epidemiology, pathophysiology, and potential therapies.
        Am J Physiol Lung Cell Mol Physiol. 2007; 292: L813-L823
        • Mehta A.J.
        • Joshi P.C.
        • Fan X.
        • et al.
        Zinc supplementation restores PU.1 and Nrf2 nuclear binding in alveolar macrophages and improves redox balance and bacterial clearance in the lungs of alcohol-fed rats.
        Alcohol Clin Exp Res. 2011; 35: 1519-1528
        • Curry-McCoy T.V.
        • Venado A.
        • Guidot D.M.
        • et al.
        Alcohol ingestion disrupts alveolar epithelial barrier function by activation of macrophage-derived transforming growth factor beta1.
        Respir Res. 2013; 14
        • Brown L.A.
        • Harris F.L.
        • Bechara R.
        • et al.
        Effect of chronic ethanol ingestion on alveolar type II cell: glutathione and inflammatory mediator-induced apoptosis.
        Alcohol Clin Exp Res. 2001; 25: 1078-1085
        • Reynaert N.L.
        • Gopal P.
        • Rutten E.P.
        • et al.
        Advanced glycation end products and their receptor in age-related, non-communicable chronic inflammatory diseases; overview of clinical evidence and potential contributions to disease.
        Int J Biochem Cell Biol. 2016;
        • Jin X.
        • Yao T.
        • Zhou Z.
        • et al.
        Advanced glycation end products enhance macrophages polarization into M1 phenotype through activating RAGE/NF-kappaB pathway.
        Biomed Res Int. 2015; 2015: 732450
        • Jabaudon M.
        • Futier E.
        • Roszyk L.
        • et al.
        Soluble form of the receptor for advanced glycation end products is a marker of acute lung injury but not of severe sepsis in critically ill patients.
        Crit Care Med. 2011; 39: 480-488
        • Fan X.
        • Joshi P.C.
        • Koval M.
        • et al.
        Chronic alcohol ingestion exacerbates lung epithelial barrier dysfunction in HIV-1 transgenic rats.
        Alcohol Clin Exp Res. 2011; 35: 1866-1875
        • Chen Y.
        • Huang X.J.
        • Yu N.
        • et al.
        HMGB1 Contributes to the expression of P-glycoprotein in mouse epileptic brain through toll-like receptor 4 and receptor for advanced glycation end products.
        PLoS One. 2015; 10: e0140918
        • Yeligar S.M.
        • Harris F.L.
        • Hart C.M.
        • et al.
        Glutathione attenuates ethanol-induced alveolar macrophage oxidative stress and dysfunction by downregulating NADPH oxidases.
        Am J Physiol Lung Cell Mol Physiol. 2014; 306: L429-L441
        • Jensen J.S.
        • Fan X.
        • Guidot D.M.
        Alcohol causes alveolar epithelial oxidative stress by inhibiting the nuclear factor (erythroid-derived 2)-like 2-antioxidant response element signaling pathway.
        Am J Respir Cell Mol Biol. 2013; 48: 511-517
        • Lois M.
        • Brown L.A.
        • Moss I.M.
        • et al.
        Ethanol ingestion increases activation of matrix metalloproteinases in rat lungs during acute endotoxemia.
        Am J Respir Crit Care Med. 1999; 160: 1354-1360
        • Mehta A.J.
        • Yeligar S.M.
        • Elon L.
        • et al.
        Alcoholism causes alveolar macrophage zinc deficiency and immune dysfunction.
        Am J Respir Crit Care Med. 2013; 188: 716-723
        • Yeligar S.M.
        • Mehta A.J.
        • Harris F.L.
        • et al.
        Peroxisome proliferator-activated receptor gamma regulates chronic alcohol-induced alveolar macrophage dysfunction.
        Am J Respir Cell Mol Biol. 2016; 55: 35-46
        • Joshi P.C.
        • Applewhite L.
        • Ritzenthaler J.D.
        • et al.
        Chronic ethanol ingestion in rats decreases granulocyte-macrophage colony-stimulating factor receptor expression and downstream signaling in the alveolar macrophage.
        J Immunol. 2005; 175: 6837-6845
        • Deane R.
        • Singh I.
        • Sagare A.P.
        • et al.
        A multimodal RAGE-specific inhibitor reduces amyloid beta-mediated brain disorder in a mouse model of Alzheimer disease.
        J Clin Invest. 2012; 122: 1377-1392
        • Uchida T.
        • Shirasawa M.
        • Ware L.B.
        • et al.
        Receptor for advanced glycation end-products is a marker of type I cell injury in acute lung injury.
        Am J Respir Crit Care Med. 2006; 173: 1008-1015