PMID- 28619997
OWN - NLM
STAT- MEDLINE
DCOM- 20170808
LR  - 20191210
IS  - 1524-4636 (Electronic)
IS  - 1079-5642 (Print)
IS  - 1079-5642 (Linking)
VI  - 37
IP  - 8
DP  - 2017 Aug
TI  - Cardiotonic Steroids Stimulate Macrophage Inflammatory Responses Through a 
      Pathway Involving CD36, TLR4, and Na/K-ATPase.
PG  - 1462-1469
LID - 10.1161/ATVBAHA.117.309444 [doi]
AB  - OBJECTIVE: Circulating levels of cardiotonic steroids (CTS) are elevated in 
      various chronic inflammatory conditions, but the role of CTS in inflammation 
      remains largely unknown. We have previously shown that the CTS ouabain stimulates 
      proinflammatory responses in murine macrophages. In this study, we aim to explore 
      the mechanism how CTS induce proinflammatory responses in primary murine and 
      human macrophages. APPROACH AND RESULTS: Using both murine peritoneal macrophages 
      and human monocyte-derived macrophages, we demonstrated that ouabain activated 
      NF-kappaB (nuclear factor kappa-light-chain-enhancer of activated B cells), leading 
      to proinflammatory cytokine (eg, MCP-1 [monocyte chemotactic protein 1], TNF-alpha 
      [tumor necrosis factor-alpha], IL-1beta [interleukin-1beta], and IL-6) production. By 
      applying siRNA techniques and murine peritoneal macrophages isolated from 
      genetically modified mice, we showed that macrophages partially deficient in 
      Na/K-ATPase, the receptor for CTS, or fully deficient in the scavenger receptor 
      CD36 or TLR4 (Toll-like receptor) were resistant to ouabain-induced NF-kappaB 
      activation, suggesting an indispensable role of these 3 receptors in this 
      pathway. Mechanistically, this effect of ouabain was independent of the ion 
      transport function of the Na/K-ATPase. Instead, ouabain stimulated a signaling 
      complex, including Na/K-ATPase, CD36, and TLR4. Subsequently, TLR4 recruited 
      MyD88 adaptor protein for NF-kappaB activation. Furthermore, intraperitoneal 
      injection of ouabain into mice specifically recruited Ly6C(+)CCR2(+) monocyte 
      subtypes to the peritoneal cavities, indicating that the CTS ouabain triggers 
      inflammation in vivo. CONCLUSIONS: CTS activate NF-kappaB leading to proinflammatory 
      cytokine production in primary macrophages through a signaling complex, including 
      CD36, TLR4, and Na/K-ATPase. These findings warrant further studies on endogenous 
      CTS in chronic inflammatory diseases, such as atherosclerosis.
CI  - (c) 2017 American Heart Association, Inc.
FAU - Chen, Yiliang
AU  - Chen Y
AD  - From the Blood Research Institute, Blood Center of Wisconsin, Milwaukee (Y.C., 
      W.H., M.Y., G.X., W.C., R.L.S.); Department of Cell Biology, Neurobiology and 
      Anatomy (M.Y., R.L.S.) and Department of Medicine (R.L.S.), Medical College of 
      Wisconsin, Milwaukee; and Departments of Medicine, Pharmacology and Surgery, Joan 
      C. Edwards School of Medicine, Marshall University, Huntington, WV (Z.X.).
FAU - Huang, Wenxin
AU  - Huang W
AD  - From the Blood Research Institute, Blood Center of Wisconsin, Milwaukee (Y.C., 
      W.H., M.Y., G.X., W.C., R.L.S.); Department of Cell Biology, Neurobiology and 
      Anatomy (M.Y., R.L.S.) and Department of Medicine (R.L.S.), Medical College of 
      Wisconsin, Milwaukee; and Departments of Medicine, Pharmacology and Surgery, Joan 
      C. Edwards School of Medicine, Marshall University, Huntington, WV (Z.X.).
FAU - Yang, Moua
AU  - Yang M
AD  - From the Blood Research Institute, Blood Center of Wisconsin, Milwaukee (Y.C., 
      W.H., M.Y., G.X., W.C., R.L.S.); Department of Cell Biology, Neurobiology and 
      Anatomy (M.Y., R.L.S.) and Department of Medicine (R.L.S.), Medical College of 
      Wisconsin, Milwaukee; and Departments of Medicine, Pharmacology and Surgery, Joan 
      C. Edwards School of Medicine, Marshall University, Huntington, WV (Z.X.).
FAU - Xin, Gang
AU  - Xin G
AD  - From the Blood Research Institute, Blood Center of Wisconsin, Milwaukee (Y.C., 
      W.H., M.Y., G.X., W.C., R.L.S.); Department of Cell Biology, Neurobiology and 
      Anatomy (M.Y., R.L.S.) and Department of Medicine (R.L.S.), Medical College of 
      Wisconsin, Milwaukee; and Departments of Medicine, Pharmacology and Surgery, Joan 
      C. Edwards School of Medicine, Marshall University, Huntington, WV (Z.X.).
FAU - Cui, Weiguo
AU  - Cui W
AD  - From the Blood Research Institute, Blood Center of Wisconsin, Milwaukee (Y.C., 
      W.H., M.Y., G.X., W.C., R.L.S.); Department of Cell Biology, Neurobiology and 
      Anatomy (M.Y., R.L.S.) and Department of Medicine (R.L.S.), Medical College of 
      Wisconsin, Milwaukee; and Departments of Medicine, Pharmacology and Surgery, Joan 
      C. Edwards School of Medicine, Marshall University, Huntington, WV (Z.X.).
FAU - Xie, Zijian
AU  - Xie Z
AD  - From the Blood Research Institute, Blood Center of Wisconsin, Milwaukee (Y.C., 
      W.H., M.Y., G.X., W.C., R.L.S.); Department of Cell Biology, Neurobiology and 
      Anatomy (M.Y., R.L.S.) and Department of Medicine (R.L.S.), Medical College of 
      Wisconsin, Milwaukee; and Departments of Medicine, Pharmacology and Surgery, Joan 
      C. Edwards School of Medicine, Marshall University, Huntington, WV (Z.X.).
FAU - Silverstein, Roy L
AU  - Silverstein RL
AD  - From the Blood Research Institute, Blood Center of Wisconsin, Milwaukee (Y.C., 
      W.H., M.Y., G.X., W.C., R.L.S.); Department of Cell Biology, Neurobiology and 
      Anatomy (M.Y., R.L.S.) and Department of Medicine (R.L.S.), Medical College of 
      Wisconsin, Milwaukee; and Departments of Medicine, Pharmacology and Surgery, Joan 
      C. Edwards School of Medicine, Marshall University, Huntington, WV (Z.X.). 
      rsilverstein@mcw.edu.
LA  - eng
GR  - P01 HL087018/HL/NHLBI NIH HHS/United States
GR  - R01 AI125741/AI/NIAID NIH HHS/United States
GR  - R01 HL109015/HL/NHLBI NIH HHS/United States
GR  - R01 HL111614/HL/NHLBI NIH HHS/United States
PT  - Journal Article
DEP - 20170615
PL  - United States
TA  - Arterioscler Thromb Vasc Biol
JT  - Arteriosclerosis, thrombosis, and vascular biology
JID - 9505803
RN  - 0 (CD36 Antigens)
RN  - 0 (Cardiotonic Agents)
RN  - 0 (Cytokines)
RN  - 0 (Inflammation Mediators)
RN  - 0 (MYD88 protein, human)
RN  - 0 (Myd88 protein, mouse)
RN  - 0 (Myeloid Differentiation Factor 88)
RN  - 0 (NF-kappa B)
RN  - 0 (TLR4 protein, human)
RN  - 0 (Tlr4 protein, mouse)
RN  - 0 (Toll-Like Receptor 4)
RN  - 5ACL011P69 (Ouabain)
RN  - EC 3.6.1.- (ATP1A1 protein, human)
RN  - EC 7.2.2.13 (Atp1a1 protein, mouse)
RN  - EC 7.2.2.13 (Sodium-Potassium-Exchanging ATPase)
SB  - IM
MH  - Animals
MH  - CD36 Antigens/deficiency/genetics/*metabolism
MH  - Cardiotonic Agents/*toxicity
MH  - Cells, Cultured
MH  - Cytokines/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Enzyme Activation
MH  - Female
MH  - Inflammation/*chemically induced/enzymology/genetics
MH  - Inflammation Mediators/*metabolism
MH  - Macrophages, Peritoneal/*drug effects/enzymology
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Knockout
MH  - Myeloid Differentiation Factor 88/metabolism
MH  - NF-kappa B/genetics/metabolism
MH  - Ouabain/*toxicity
MH  - RNA Interference
MH  - Signal Transduction/drug effects
MH  - Sodium-Potassium-Exchanging ATPase/deficiency/genetics/*metabolism
MH  - Time Factors
MH  - Toll-Like Receptor 4/*metabolism
MH  - Transfection
PMC - PMC5532064
MID - NIHMS881990
OTO - NOTNLM
OT  - atherosclerosis
OT  - cytokines
OT  - inflammation
OT  - ion pumps
OT  - macrophages
EDAT- 2017/06/18 06:00
MHDA- 2017/08/09 06:00
PMCR- 2018/08/01
CRDT- 2017/06/17 06:00
PHST- 2017/01/04 00:00 [received]
PHST- 2017/06/01 00:00 [accepted]
PHST- 2017/06/18 06:00 [pubmed]
PHST- 2017/08/09 06:00 [medline]
PHST- 2017/06/17 06:00 [entrez]
PHST- 2018/08/01 00:00 [pmc-release]
AID - ATVBAHA.117.309444 [pii]
AID - 10.1161/ATVBAHA.117.309444 [doi]
PST - ppublish
SO  - Arterioscler Thromb Vasc Biol. 2017 Aug;37(8):1462-1469. doi: 
      10.1161/ATVBAHA.117.309444. Epub 2017 Jun 15.