Triptolide is a potent suppressant of C3, CD40 and B7h expression in activated human proximal tubular epithelial cells.
BACKGROUND
Previous studies have shown that triptolide possesses potent immunosuppressive and anti-inflammatory properties. Increasing recognition of the importance of the proximal tubular epithelial cells (PTEC) in renal disease and renal transplantation raises the question of whether triptolide suppresses the pro-inflammatory activity of PTEC.


METHODS
Cultured human PTEC were exposed to tumor necrosis factor-alpha (TNF-alpha) and immunosuppressant (triptolide or CsA or FK506) for 24 hours, followed by RT-PCR, ELISA, flow cytometry and Western blotting analysis for complement C3, CD40, B7h expression.


RESULTS
TNF-alpha up-regulated C3, CD40 and B7h production by PTEC. This up-regulation was inhibited by all three immunosuppressants with different intensity. Firstly, triptolide (4 to 8 ng/mL), CsA (4000 to 6000 ng/mL) and FK506 (2000 ng/mL) inhibited up-regulation of C3 mRNA, but CsA and FK506 had less of an effect than triptolide. Secondly, triptolide (4 to 8 ng/mL) completely inhibited C3 expression at both mRNA and protein levels. In contrast, CsA and FK506 had only slight effects on C3 expression at the protein level. Thirdly, triptolide (4 to 8 ng/mL), CsA (500 to 2500 ng/mL) and FK506 (1250 ng/mL) inhibited up-regulation of CD40 and B7h mRNA, the effect on B7h and CD40 mRNA expression by CsA and FK506 being greater than that on C3 mRNA expression.


CONCLUSIONS
Triptolide effectively inhibited up-regulation of C3, CD40 and B7h on PTEC. Triptolide was more effective than CsA and FK506 at inhibiting C3 expression. This suggests that triptolide, at non-cytotoxic concentrations, has the potential to reduce the inflammatory and immunostimulatory properties of PTEC, in addition to any of the previously reported actions on T cell or B cell function.
Hong
Yuzhi
Y
Department of Nephrology and Transplantation, Guy's Hospital, London SE1 9RT, England, United Kingdom.
Zhou
Wuding
W
Li
Ke
K
Sacks
Steven H
SH
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology* / Q000633:toxicity; D015703:Antigens, CD; D000970:Antineoplastic Agents / Q000494:pharmacology; D019013:CD40 Antigens / Q000235:genetics*; D003176:Complement C3 / Q000235:genetics* / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology / Q000633:toxicity; D004224:Diterpenes / Q000494:pharmacology* / Q000633:toxicity; D004852:Epoxy Compounds; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology / Q000633:toxicity; D060945:Inducible T-Cell Co-Stimulator Ligand; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000502:physiology*; D010616:Phenanthrenes; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D016559:Tacrolimus / Q000494:pharmacology / Q000633:toxicity; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D015703:Antigens, CD; D000970:Antineoplastic Agents; D019013:CD40 Antigens; D003176:Complement C3; D004224:Diterpenes; D004852:Epoxy Compounds; C498514:ICOSLG protein, human; D007166:Immunosuppressive Agents; D060945:Inducible T-Cell Co-Stimulator Ligand; D010616:Phenanthrenes; D011506:Proteins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C001899:triptolide; D016572:Cyclosporine; D016559:Tacrolimus