Mechanism of altered TNF-α expression by macrophage and the modulatory effect of Panax notoginseng saponins in scald mice

• Published in: Burns Vol. 32; no. 7; pp. 846 - 852
• Main Authors: Wang, Yong, Peng, Daizhi, Huang, Wenhua, Zhou, Xin, Liu, Jin, Fang, Yongfei
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.11.2006; Elsevier Science
• Subjects: Animals; Biological and medical sciences; Blotting, Western; Burns; Burns - metabolism; Electrophoresis; Female; Macrophage; Macrophages - metabolism; Male; Medical sciences; Mice; NF-kappa B - metabolism; Nuclear factor-κB; Panax notoginseng; Protein kinase C; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Saponins - pharmacology; Traumas. Diseases due to physical agents; Tumor necrosis factor; Tumor Necrosis Factor-alpha - metabolism; Protein kinase C; Burns; Nuclear factor-κB; Tumor necrosis factor; Macrophage; Skin disease; Enzyme; Transferases; Cytokine; Rodentia; Scald; Mechanism; Burn; Panax notoginseng; Vertebrata; Mammalia; Mouse; Dicotyledones; Animal; Angiospermae; Spermatophyta; Nuclear factor-KB; Araliaceae; Tumor necrosis factor α
• ISSN: 0305-4179; 1879-1409
• DOI: 10.1016/j.burns.2006.02.001

To explore the mechanism of altered tumor necrosis factor-alpha (TNF-α) expression by peritoneal macrophages (PMΦ) and Panax notoginseng saponins (PNS) modulation in light of NF-κB signal transduction in severely scalded mice. Eighteen percent total body surface area (TBSA) full-thickness scalded mice were used. PMΦ was collected at different time intervals (0, 2, 6, 12, 24 and 48 post-burn hour (PBH)) separately. The following parameters were measured: TNF-α mRNA and IL-10 mRNA expression (reverse transcription-polymerase chain reaction, RT-PCR), protein kinase C (PKC) activity (isotope incorporation analysis), NF-κB activity (electrophoretic mobility shift assay, EMSA), IκB-α expression (Western blot). After scald, increased expression of TNF-α mRNA of PMΦ peaked at 12 PBH. Meanwhile, expression of IL-10 mRNA dropped to the lowest level at 12 PBH. NF-κB activity was markedly activated and reached its peak at 2 PBH. Membrane PKC activity was up-regulated after scald and showed a positive correlation with the change of TNF-α mRNA. Expression of IκB-α first decreased at 2 PBH and then increased to high level at 24 PBH. When 12 PBH was chosen as the time point for in vitro intervention with the application of specific NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC), PKC inhibitor H-7 and PNS, both TNF-α mRNA expression and NF-κB activity decreased significantly. These results indicate that abnormal expression of TNF-α mRNA of macrophages might be regulated by PKC-NF-κB signaling following severe burn. PNS might play an anti-inflammatory effect by inhibiting NF-κB activity and TNF-α mRNA expression.