title: Berberine alleviates ox-LDL induced inflammatory factors by up-regulation of autophagy via AMPK/mTOR signaling pathway.
abstract: BACKGROUND
  Inflammation induced by oxidized low-density lipoprotein (ox-LDL) plays an important role in the pathogenesis of atherosclerosis. Recently, roles of autophagy against inflammation in the process of atherosclerosis have drawn increasing attention. Here, we tested the possible molecular mechanisms by which berberine confers an anti-inflammatory effect in macrophages by upregulation of autophagy.
  
  
  METHODS
  J774A.1 macrophages were incubated with various doses of ox-LDL for various times. We evaluated the inflammatory factors and autophagy proteins (LC3II/LC3I, and SQSTM1/p62) to ascertain the optimal dose and time. Ox-LDL-induced inflammatory factors and autophagy in J774A.1 cells were tested by the AimPlex multiplex assay, Western blotting, confocal microscopy, and transmission electron microscopy in the presence of berberine or chloroquine (CQ). Adenosine 5'-monophosphate-activated protein kinase (AMPK) inhibitor compound C was used to evaluate the AMPK/mTOR signaling pathway.
  
  
  RESULTS
  Berberine dose- and time-dependently reduced ox-LDL-induced inflammation and increased the ratio of LC3II/LC3I, and SQSTM1/p62 in J774A.1 cells. CQ significantly attenuated the berberine-induced autophagy and anti-inflammation. In addition, berberine increased the ratio of p-AMPK/AMPK and decreased the ratio of p-mTOR/mTOR. AMPK inhibitor compound C abolished berberine-induced autophagy and promoted p-mTOR/mTOR expression in J774A.1 cells.
  
  
  CONCLUSIONS
  Berberine treatment inhibits inflammation in J774A.1 cells by inducing autophagy, which is mediated through activation of the AMPK/mTOR signaling pathway. Importantly, this study provides new insight into berberine's molecular mechanism and its therapeutic potential in the treatment of atherosclerosis.
authors.affiliation: Department of Tianjin, University of Traditional Chinese Medicine, Tianjin, 300193, China. fanxdfanxd@126.com.|Department of Experimental Research Center, Xiyuan Hospital of China Academy of Chinese Medical Sciences, 1 Xiyuan Caochang, Hai Dian District, Beijing, 100091, China. fanxdfanxd@126.com.
authors.forename: Xiaodi
authors.initials: X
authors.lastname: Fan
authors.affiliation: Institute of Basic Theory, China Academy of Chinese Medical Sciences, Beijing, 100700, China. maggie.wj@126.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Experimental Research Center, Xiyuan Hospital of China Academy of Chinese Medical Sciences, 1 Xiyuan Caochang, Hai Dian District, Beijing, 100091, China. houjchoujc@126.com.
authors.forename: Jincai
authors.initials: J
authors.lastname: Hou
authors.affiliation: Department of Experimental Research Center, Xiyuan Hospital of China Academy of Chinese Medical Sciences, 1 Xiyuan Caochang, Hai Dian District, Beijing, 100091, China. lincrlincr@126.com.
authors.forename: Chengren
authors.initials: C
authors.lastname: Lin
authors.affiliation: Centre for Complementary Medicine Research, School of Science and Health, University of Western Sydney, Parramatta, NSW, 2751, Australia. wang_liru2009@126.com.
authors.forename: Alan
authors.initials: A
authors.lastname: Bensoussan
authors.affiliation: Centre for Complementary Medicine Research, School of Science and Health, University of Western Sydney, Parramatta, NSW, 2751, Australia. maggiemaggie1979@163.com.
authors.forename: Dennis
authors.initials: D
authors.lastname: Chang
authors.affiliation: Department of Experimental Research Center, Xiyuan Hospital of China Academy of Chinese Medical Sciences, 1 Xiyuan Caochang, Hai Dian District, Beijing, 100091, China. ljxunljxun@163.com.
authors.forename: Jianxun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Anesthesiology and Critical Care Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA. wangbwangb@126.com.
authors.forename: Bing
authors.initials: B
authors.lastname: Wang
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D000818:Animals; D001343:Autophagy / Q000187:drug effects*; D001599:Berberine / Q000494:pharmacology*; D002460:Cell Line; D054405:Chemokine CCL3 / Q000378:metabolism; D018946:Chemokine CCL5 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015151:Immunoblotting; D018836:Inflammation Mediators / Q000378:metabolism*; D016753:Interleukin-10 / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D013997:Time Factors; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: C503460:Ccl5 protein, mouse; D054405:Chemokine CCL3; D018946:Chemokine CCL5; D018836:Inflammation Mediators; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D001599:Berberine; D016753:Interleukin-10; D058570:TOR Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases