title: 13-Methylberberine reduces HMGB1 release in LPS-activated RAW264.7 cells and increases the survival of septic mice through AMPK/P38 MAPK activation.
abstract: High mobility group box 1 (HMGB1), a late phase cytokine of sepsis, is viewed as a potential target for the treatment of sepsis. The authors considered that 13-methylberberine (13-MB) might reduce circulating HMGB1 levels and increase survival in a mouse model of sepsis by activating AMP-activated protein kinase (AMPK). Western blot analysis and vascular contraction testing were performed using RAW264.7 cells and rat thoracic aorta, respectively. The mechanisms responsible were investigated using various signal inhibitors and small interfering RNA techniques. 13-MB significantly reduced HMGB1 release by LPS-activated RAW264.7 cells, and this was prevented by silencing AMPK or p38, or by pretreating cells with p38 MAPKinase inhibitor, suggesting that the activations of p38 and AMPK were responsible for the observed reduction in HMGB1 release. As was expected, 13-MB increased the phosphorylations of p38 and AMPK. Interestingly, phosphorylations of p38 by 13-MB were inhibited by AMPKsiRNA, indicating that AMPK lies upstream of p38. Furthermore, 13-MB concentration-dependently inhibited IκB phosphorylation in LPS-activated RAW264.7 cells, and in aortic rings, co-treatment with 13-MB and LPS for 8h, in vitro, significantly restored the isometric contraction induced by phenylephrine. Importantly, 13-MB significantly increased the survival rate of LPS-induced endotoxemic mice. These results suggest 13-MB may be useful for treating diseases in which HMGB1 is viewed as a target.
authors.affiliation: Department of Pharmacology, School of Medicine Gyeongsang National University and Institute of Health Sciences, Jinju 660-751, Republic of Korea.
authors.forename: Ki Churl
authors.initials: KC
authors.lastname: Chang
authors.affiliation: Department of Pharmacology, School of Medicine Gyeongsang National University and Institute of Health Sciences, Jinju 660-751, Republic of Korea.
authors.forename: Young Shin
authors.initials: YS
authors.lastname: Ko
authors.affiliation: Department of Pharmacology, School of Medicine Gyeongsang National University and Institute of Health Sciences, Jinju 660-751, Republic of Korea.
authors.forename: Hye Jung
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Division of Bioscience, Dongguk University, Gyeongju 780-714, Republic of Korea.
authors.forename: Da-Yeong
authors.initials: DY
authors.lastname: Nam
authors.affiliation: Division of Bioscience, Dongguk University, Gyeongju 780-714, Republic of Korea. Electronic address: dulee@dongkuk.ac.kr.
authors.forename: Dong-Ung
authors.initials: DU
authors.lastname: Lee
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000502:physiology; D001599:Berberine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000627:therapeutic use; D024243:HMGB1 Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism; D066298:In Vitro Techniques; D008070:Lipopolysaccharides; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism; D000067996:RAW 264.7 Cells; D034741:RNA, Small Interfering / Q000235:genetics; D017207:Rats, Sprague-Dawley; D018805:Sepsis / Q000188:drug therapy*; D015398:Signal Transduction / Q000187:drug effects; D014661:Vasoconstriction / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D024243:HMGB1 Protein; C582808:HMGB1 protein, mouse; D008070:Lipopolysaccharides; D016328:NF-kappa B; D034741:RNA, Small Interfering; D001599:Berberine; D008156:Luciferases; D048051:p38 Mitogen-Activated Protein Kinases; D055372:AMP-Activated Protein Kinases
keywords: 13-methylberberine; AMPK; HMGB1; Inflammation; Sepsis; p38MAPK