title: [Triptolide inhibits the inflammatory response of monocytes from rheumatoid arthritis patients by regulating miR-155].
abstract: OBJECTIVE
  To explore the anti-inflammatory effect of triptolide (TPT) by regulating miR-155 in monocytes pre-stimulated by lipopolysaccharide (LPS) from rheumatoid arthritis (RA) patients.
  
  
  METHODS
  Monocytes were isolated by CD14⁺ magnetic beads from peripheral blood mononuclear cells (PBMCs) of RA and stimulated by LPS for 24 hours. The levels of tumor-necrosis factor-α (TNF-α) and interleukin 6 (IL-6) in monocytes were detected by ELISA and the expression of miR-155 was measured by real-time quantitative PCR (qRT-PCR) in monocytes before and after the treatment of TPT at different concentrations. MiR-155 mimic and negative control were respectively transfected into the LPS-stimulated monocytes by Lipofectamine(TM)2000. Twenty-four hours later, the monocytes were treated with or without TPT for another 24 hours. TNF-α and IL-6 expressions in the cell culture supernatants were detected by ELISA and the expressions of suppressor of cytokine signaling-1 (SOCS1) and Src homology 2 domain-containing inositol 5-phosphatase 1 (SHIP-1) were tested by Western blotting.
  
  
  RESULTS
  TPT suppressed the expressions of TNF-α, IL-6 and miR-155 in LPS-stimulated peripheral blood monocytes from RA patients. Over-expression of miR-155 significantly reversed the down-regulation of TNF-α and IL-6 by TPT in monocytes. TPT up-regulated the expressions of SOCS1 and SHIP-1 in monocytes, but over-expressed miR-155 antagonized the effect of TPT on SHIP-1 while the expression of SOCS1 was not affected.
  
  
  CONCLUSIONS
  TPT suppressed the expression of miR-155 and up-regulated the release of SHIP-1, thus inhibiting the inflammatory response in the LPS-stimulated monocytes of RA patients.
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
authors.forename: Anping
authors.initials: A
authors.lastname: Peng
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
authors.forename: Xianzhang
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
authors.forename: Ruiping
authors.initials: R
authors.lastname: Liu
authors.affiliation: Department of Gynecology, Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
authors.forename: Junhua
authors.initials: J
authors.lastname: Zhuang
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D001172:Arthritis, Rheumatoid / Q000097:blood* / Q000235:genetics / Q000473:pathology; D015153:Blotting, Western; D002478:Cells, Cultured; D004224:Diterpenes / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D004852:Epoxy Compounds / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D000072181:Inositol Polyphosphate 5-Phosphatases; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D035683:MicroRNAs / Q000235:genetics*; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010616:Phenanthrenes / Q000494:pharmacology*; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000378:metabolism; D029757:Tripterygium / Q000737:chemistry; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D004224:Diterpenes; D004852:Epoxy Compounds; D015850:Interleukin-6; D008070:Lipopolysaccharides; C524942:MIRN155 microRNA, human; D035683:MicroRNAs; D010616:Phenanthrenes; C108944:SOCS1 protein, human; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D014409:Tumor Necrosis Factor-alpha; C001899:triptolide; D010744:Phosphoric Monoester Hydrolases; D000072181:Inositol Polyphosphate 5-Phosphatases; C000606430:INPP5D protein, human; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases