title: Effect of CD95 on inflammatory response in rheumatoid arthritis fibroblast-like synoviocytes.
abstract: OBJECTIVE
  Many CD95-expressing cells don't always undergo apoptosis after stimulation with CD95 ligation. The purpose of this paper is to investigate the role of expression of CD95 (Fas/Apo1) on inflammatory response in fibroblast-like synoviocytes (FLS) obtained from rheumatoid arthritis (RA) and to evaluate the role of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB or Akt) pathways within this process.
  
  
  METHODS
  The expression levels of CD95 were monitored by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). Apoptotic cells were detected by in situ apoptosis detection (TUNEL) assay. The RA-FLS were treated with agonistic anti-CD95 antibody or CD95 siRNA. Then the proliferation was detected by CCK-8, and mRNA level of inflammatory cytokines was detected by RT-PCR. After the RA-FLS were treated with agonistic anti-CD95 antibody, the total Akt and pAkt protein expression was analyzed by Western blot, and the changes mentioned above were observed while pre-incubated with the PI3K inhibitor LY294002.
  
  
  RESULTS
  A significant increase of CD95 antigen was found in RA compared with osteoarthritis (OA) samples, while apoptosis in RA synovial tissue was not obvious. Low concentrations of agonistic anti-CD95 antibody could promote RA-FLS growth and interleukin-6 (IL-6) mRNA expression, while high concentrations could induce apoptosis. And both of these phenomena could be inhibited by CD95 siRNA. Agonistic anti-CD95 antibody could stimulate the expression of pAkt, and PI3K specific inhibitor LY294002 could induce opposite changes.
  
  
  CONCLUSIONS
  Stimulation of CD95 could promote RA-FLS proliferation and inflammation, and activation of the PI3K/Akt signaling pathway might be the possible mechanism.
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Xiaoqiong
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Zhanfeng
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Anping
authors.initials: A
authors.lastname: Peng
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Min
authors.initials: M
authors.lastname: He
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Xu
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Sujing
authors.initials: S
authors.lastname: Shen
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China.
authors.forename: Junhua
authors.initials: J
authors.lastname: Zhuang
authors.affiliation: Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, PR China. Electronic address: huangxz020@163.com.
authors.forename: Xianzhang
authors.initials: X
authors.lastname: Huang
mesh_terms: D017209:Apoptosis / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D005347:Fibroblasts / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D007249:Inflammation / Q000276:immunology* / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000276:immunology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000276:immunology; D013583:Synovial Membrane / Q000276:immunology* / Q000378:metabolism; D019014:fas Receptor / Q000276:immunology* / Q000378:metabolism
chemical_list: C491901:FAS protein, human; D019014:fas Receptor; D051057:Proto-Oncogene Proteins c-akt
keywords: CD95; Inflammatory; Proliferation; Rheumatoid arthritis; Synoviocyte