PMID- 31100475 OWN - NLM STAT- MEDLINE DCOM- 20200615 LR - 20200615 IS - 1873-4596 (Electronic) IS - 0891-5849 (Linking) VI - 139 DP - 2019 Aug 1 TI - The extracellular role of Ref-1 as anti-inflammatory function in lipopolysaccharide-induced septic mice. PG - 16-23 LID - S0891-5849(19)30442-3 [pii] LID - 10.1016/j.freeradbiomed.2019.05.013 [doi] AB - Apurinic/apyrimidinic endonuclease/redox factor-1 (Ref-1), a multifunctional protein secreted from stimulated cells, has been identified as a new serological biomarker. Despite recent reports on the role of Ref-1 in inflammation, the biological function of secreted Ref-1 remains unknown, especially in vivo. This study aimed to evaluate the possible roles of secreted Ref-1 in lipopolysaccharide-induced systemic inflammation in vivo. We generated a secretory Ref-1 adenoviral vector system, AdPPT-LS-Ref-1, by conjugation of preprotrypsin leading sequence (PPT-LS) with full-length Ref-1 sequences. Expression of tumor necrosis factor-alpha (TNF-alpha)-induced vascular cell adhesion molecule-1 (VCAM-1) in endothelial cells and lipopolysaccharide (LPS)-induced cyclooxygenase-2 in Raw264.7 cells was inhibited by secretory Ref-1, and this inhibitory effect was abrogated following neutralization of Ref-1 with anti-Ref-1 antibody. Plasma Ref-1 levels following administration of AdPPT-LS-Ref-1 (2 x 10(9) ifu, i.p.) for 24 h were substantially higher than those recorded following administration of Adbetagal (84.6 +/- 7.2 ng/ml vs. 4.4 +/- 1.5 ng/ml). Treatment with LPS (10 mg/kg, i.v. for 6 h) markedly increased VCAM-1 expression, cathepsin or myeloperoxidase activity, which were significantly suppressed by treatment with AdPPT-LS-Ref-1. Furthermore, LPS-induced cytokines, such as TNF-alpha, interleukin (IL)-1beta, IL-6, and monocyte chemoattractant protein 1, were significantly inhibited in AdPPT-LS-Ref-1-treated mice. However, LPS-induced myeloperoxidase activities were not suppressed by treatment with the redox mutant of secretory Ref-1, AdPPT-LS-Ref-1(C65A/C93A), or wild-type AdRef-1. Collectively, these results suggest that secreted Ref-1 has anti-inflammatory properties and that its redox cysteine residue is associated with the anti-inflammatory activity in vivo. Furthermore, our findings indicate that secretory Ref-1 may be useful as a therapeutic biomolecule against systemic inflammation. CI - Copyright (c) 2019 Elsevier Inc. All rights reserved. FAU - Joo, Hee Kyoung AU - Joo HK AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. FAU - Lee, Yu Ran AU - Lee YR AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. FAU - Lee, Eun-Ok AU - Lee EO AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. FAU - Park, Myoung Soo AU - Park MS AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. FAU - Choi, Sunga AU - Choi S AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. FAU - Kim, Cuk-Seong AU - Kim CS AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. FAU - Park, Jin-Bong AU - Park JB AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. FAU - Jeon, Byeong Hwa AU - Jeon BH AD - Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon, South Korea. Electronic address: bhjeon@cnu.ac.kr. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190515 PL - United States TA - Free Radic Biol Med JT - Free radical biology & medicine JID - 8709159 RN - 0 (Anti-Inflammatory Agents, Non-Steroidal) RN - 0 (Ccl2 protein, mouse) RN - 0 (Chemokine CCL2) RN - 0 (IL1B protein, mouse) RN - 0 (Interleukin-1beta) RN - 0 (Interleukin-6) RN - 0 (Lipopolysaccharides) RN - 0 (Protein Precursors) RN - 0 (Tumor Necrosis Factor-alpha) RN - 0 (Vascular Cell Adhesion Molecule-1) RN - 0 (interleukin-6, mouse) RN - EC 1.11.1.7 (Peroxidase) RN - EC 1.14.99.- (Ptgs2 protein, mouse) RN - EC 1.14.99.1 (Cyclooxygenase 2) RN - EC 3.4.- (Cathepsins) RN - EC 3.4.21.4 (Trypsin) RN - EC 4.2.99.18 (Apex1 protein, mouse) RN - EC 4.2.99.18 (DNA-(Apurinic or Apyrimidinic Site) Lyase) SB - IM MH - Adenoviridae/genetics/metabolism MH - Animals MH - Anti-Inflammatory Agents, Non-Steroidal/metabolism/*pharmacology MH - Cathepsins/genetics/metabolism MH - Chemokine CCL2/genetics/metabolism MH - Cyclooxygenase 2/genetics/metabolism MH - DNA-(Apurinic or Apyrimidinic Site) Lyase/antagonists & inhibitors/*genetics/metabolism/*pharmacology MH - Gene Expression Regulation MH - Genetic Vectors/chemistry/metabolism MH - Human Umbilical Vein Endothelial Cells/cytology/drug effects/metabolism MH - Humans MH - Interleukin-1beta/genetics/metabolism MH - Interleukin-6/genetics/metabolism MH - Lipopolysaccharides/administration & dosage/*antagonists & inhibitors MH - Male MH - Mice MH - Mice, Inbred ICR MH - Peroxidase/genetics/metabolism MH - Protein Precursors/genetics/metabolism MH - RAW 264.7 Cells MH - Sepsis/chemically induced/genetics/pathology/*therapy MH - Trypsin/genetics/metabolism MH - Tumor Necrosis Factor-alpha/antagonists & inhibitors/genetics/metabolism/pharmacology MH - Vascular Cell Adhesion Molecule-1/genetics/metabolism EDAT- 2019/05/18 06:00 MHDA- 2020/06/17 06:00 CRDT- 2019/05/18 06:00 PHST- 2019/03/21 00:00 [received] PHST- 2019/05/11 00:00 [revised] PHST- 2019/05/11 00:00 [accepted] PHST- 2019/05/18 06:00 [pubmed] PHST- 2020/06/17 06:00 [medline] PHST- 2019/05/18 06:00 [entrez] AID - S0891-5849(19)30442-3 [pii] AID - 10.1016/j.freeradbiomed.2019.05.013 [doi] PST - ppublish SO - Free Radic Biol Med. 2019 Aug 1;139:16-23. doi: 10.1016/j.freeradbiomed.2019.05.013. Epub 2019 May 15.