title: The pentacyclic triterpene Lupeol switches M1 macrophages to M2 and ameliorates experimental inflammatory bowel disease.
abstract: BACKGROUND
  Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn's disease, is a chronic inflammatory disease in the lower gastrointestinal tract. Mounting evidence suggests that the predominance of the classically activated (M1) macrophages versus the alternatively activated (M2) macrophages plays a role in the progression of IBD. Thus, agents able to shift pro-inflammatory M1 macrophages to anti-inflammatory M2 macrophages may be beneficial to IBD. The pentacyclic triterpene Lup-20(29)-en-3β-ol (Lupeol), a potent anti-inflammatory natural product, has been shown to inhibit pro-inflammatory cytokine production, suggesting it is potentially able to modulate macrophage polarization, thereby beneficial to IBD.
  
  
  METHODS
  CD4(+) monocytes were differentiated to M1 or M2 macrophages, which were cocultured with epithelial cell lines, T84 and Caco-2, in the absence or presence of Lupeol (10μM). Experimental colitis was induced with dextran sodium sulfate (DSS), with or without oral administration of Lupeol (50mg/kg, q.d.). Cytokines were measured with Luminex kits. M1/M2 genes were measured with real-time polymerase chain reaction. Macrophage phenotypes were defined by measuring M1 and M2 markers with confocal microscopy. Proteins were measured with Western blotting, while cell surface markers were measured with confocal microscopy or flow cytometry. Histology was evaluated with H&E staining.
  
  
  RESULTS
  Treatment of M1 macrophages with Lupeol resulted in a marked decrease in the production of pro-inflammatory cytokines, including IL-12, IL6, IL-1β and TNFα, and a marked increase in the production of IL-10, an anti-inflammatory cytokine. This was associated with a down-regulation of CD86, a typical marker of M1 macrophages, and an up-regulation of CD206, a typical M2 macrophage marker. IRF5, a transcription factor that is critically involved in M1 polarization, was down-regulated in M1 macrophages after being incubated with Lupeol, associated with a marked decrease in the phosphorylation of p38 mitogen activated protein kinase. Coculture of epithelial cells with M1 macrophages resulted in down-regulation of the tight junction protein ZO-1 and disruption of epithelial integrity, which were blocked by Lupeol treatment of the M1 macrophages. Moreover, oral administration of Lupeol to dextran sulfate sodium (DSS)-induced colitis mice resulted in mitigated intestinal inflammation and increased survival from lethal colitis, associated with decreased expression of M1-related genes and increased expression of M2-related genes.
  
  
  CONCLUSIONS
  Lupeol ameliorates experimental inflammatory bowel disease through, at least in part, inhibiting M1 and promoting M2 macrophages.
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: zhuyeshan1965@163.com.
authors.forename: Yeshan
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Department of Acupuncture, North China University of Science and Technology School of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: 15531558238@163.com.
authors.forename: Xueqing
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Traditional Chinese Medicine, Yutian Hospital, Tangshan, Hebei Province, China. Electronic address: 741094776@qq.com.
authors.forename: Jianquan
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: tongjunchen@sina.com.
authors.forename: Tongjun
authors.initials: T
authors.lastname: Chen
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: shizhimin0613@163.com.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Shi
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: eimiaona001@sina.com.
authors.forename: Miaona
authors.initials: M
authors.lastname: Lei
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: zhangyj1312@126.com.
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: 854951455@qq.com.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Bai
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: liyifang02@163.com.
authors.forename: Yifang
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Digestive Internal Medicine, Tangshan Hospital of Traditional Chinese Medicine, Tangshan, Hebei Province, China. Electronic address: 1tsfuyou@126.com.
authors.forename: Xuan
authors.initials: X
authors.lastname: Fei
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003092:Colitis / Q000139:chemically induced / Q000188:drug therapy*; D016207:Cytokines / Q000378:metabolism; D016264:Dextran Sulfate / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000188:drug therapy* / Q000276:immunology; D050835:Interferon Regulatory Factors / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D053978:Pentacyclic Triterpenes / Q000008:administration & dosage*; D058408:Th1-Th2 Balance / Q000187:drug effects; D062826:Zonula Occludens-1 Protein / Q000235:genetics / Q000378:metabolism; D000094042:Lupanes
chemical_list: D016207:Cytokines; D016264:Dextran Sulfate; D050835:Interferon Regulatory Factors; D053978:Pentacyclic Triterpenes; D062826:Zonula Occludens-1 Protein; C498178:Irf5 protein, mouse; C569043:Tjp1 protein, mouse; C010480:lupeol; D000094042:Lupanes
keywords: Cytokines; Epithelial cells; Inflammatory bowel disease; Lupeol; Macrophages; Phenotypes