Anti-inflammatory constituents isolated from Clerodendron trichotomum Tunberg Leaves (CTL) inhibits pro-inflammatory gene expression in LPS-stimulated RAW 264.7 macrophages by suppressing NF-kappaB activation.
Clerodendron trichotomum Tunberg Leaves (CTL) have been used for centuries in Chinese folk medicine for their anti-inflammatory properties. To investigate the molecular mechanism of anti-inflammation by CTL, we analyzed the regulation of TNF-alpha expression in RAW 264.7 cells, a key step in inflammation. The effect of CTL on the production and expression of tumor necrosis factor-alpha (TNF-alpha) was determined by enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR). CTL inhibited the production and expression of TNF-alpha in LPS-stimulated RAW 264.7 cells in a dose-dependent manner. In addition, activation of NF-kappaB, which controls TNF-alpha expression, was inhibited in LPS-stimulated RAW 264.7 cells by CTL in a dose-dependent manner, as demonstrated by an electro phoretic mobility shift assay (EMSA). Furthermore, CTL inhibited activation of NF-kappaB through inhibition IkappaB degradation, as demonstrated by an western blot analysis of IkappaB-alpha. These results suggest that CTL inhibits the expression of the pro-inflammation gene through the inhibition of NF-kappaB dependent pathway in RAW 264.7 cells.
College of Pharmacy, Chung-Ang University, Seoul 156-756, Korea.
Min-A
MA
Park
Hong-Jin
HJ
Kim
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000302:isolation & purification / Q000494:pharmacology*; D015153:Blotting, Western; D002460:Cell Line; D032409:Clerodendrum / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay; D004797:Enzyme-Linked Immunosorbent Assay; D007249:Inflammation / Q000235:genetics*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000819:agonists*; D018515:Plant Leaves / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D000894:Anti-Inflammatory Agents, Non-Steroidal; D008070:Lipopolysaccharides; D016328:NF-kappa B; D013778:Tetrazolium Salts; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha; C022616:thiazolyl blue