Triptolide, an active compound identified in a traditional Chinese herb, induces apoptosis of rheumatoid synovial fibroblasts.
BACKGROUND
Extracts of Tripterygium wilfordii Hook F (TWHF), a traditional Chinese herb, have been reported to show efficacy in patients with rheumatoid arthritis (RA). Since RA is not only characterized by inflammation but also by synovial proliferation in the joints, we examined whether triptolide (a constituent of TWHF) could influence the proliferation of rheumatoid synovial fibroblasts (RSF) by induction of apoptosis.


RESULTS
RSF were obtained from RA patients during surgery and were treated with triptolide under various conditions. The viability and proliferation of RSF were measured by the 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) assay and by 5-bromo-2'-deoxyuridine incorporation, respectively. Apoptosis was identified by detection of DNA fragmentation using an enzyme-linked immunosorbent assay and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL). The role of caspases in apoptosis of RSF was analyzed by measuring caspase-3 activity. Activation of the peroxisome proliferator-activated receptor (PPAR) gamma was assessed by a luciferase reporter gene assay using RSF transfected with a plasmid containing the peroxisome proliferator response element. Triptolide decreased viability, inhibited proliferation, and induced apoptosis of RSF in a concentration-dependent manner at very low (nM) concentrations. Caspase-3 activity was increased by treatment with triptolide and was suppressed by caspase inhibitors. Although PPARgamma activation was induced by 15-deoxy-Delta12,14-prostaglandin J2, triptolide did not induce it under the same experimental conditions. An extract of TWHF also induced DNA fragmentation in RSF.


CONCLUSIONS
The mechanism of action remains to be studied; however, triptolide may possibly have a disease-modifying effect in patients with RA.
Kusunoki
Natsuko
N
Institute of Medical Science, St, Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki-shi, Kanagawa 216-8512, Japan. kusunoki@marianna-u.ac.jp
Yamazaki
Ryuta
R
Kitasato
Hidero
H
Beppu
Moroe
M
Aoki
Haruhito
H
Kawai
Shinichi
S
D017209:Apoptosis; D001172:Arthritis, Rheumatoid / Q000188:drug therapy / Q000473:pathology*; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004224:Diterpenes / Q000494:pharmacology* / Q000627:therapeutic use; D004365:Drugs, Chinese Herbal; D004852:Epoxy Compounds; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D010616:Phenanthrenes / Q000494:pharmacology* / Q000627:therapeutic use; D013583:Synovial Membrane / Q000187:drug effects*; D029757:Tripterygium / Q000737:chemistry
D004224:Diterpenes; D004365:Drugs, Chinese Herbal; D004852:Epoxy Compounds; D010616:Phenanthrenes; C001899:triptolide; D020169:Caspases