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

被引:37
作者
Wang, Yong
Peng, Daizhi [1 ]
Huang, Wenhua
Zhou, Xin
Liu, Jin
Fang, Yongfei
机构
[1] Third Mil Med Univ, Southwest Hosp, Inst Burn Res, State Key Lab Trauma Burs & Combined Injury, Chongqing 400038, Peoples R China
[2] Third Mil Med Univ, Southwest Hosp, Dept Tradit Chinese Med, Chongqing 400038, Peoples R China
基金
中国国家自然科学基金;
关键词
burns; tumor necrosis factor; macrophage; protein kinase C; nuclear factor-kappa B;
D O I
10.1016/j.burns.2006.02.001
中图分类号
R4 [临床医学];
学科分类号
1002 ; 100602 ;
摘要
Aim: To explore the mechanism of altered tumor necrosis factor-alpha (TNF-alpha) expression by peritoneal macrophages (PM Phi) and Panax notoginseng saponins (PNS) modulation in light of NF-kappa B signal transduction in severely scalded mice. Methods: Eighteen percent total body surface area (TBSA) full-thickness scalded mice were used. PM Phi was collected at different time intervals (0, 2, 6, 12, 24 and 48 post-burn hour (PBH)) separately. The following parameters were measured: TNF-alpha mRNA and IL-10 mRNA expression (reverse transcription-polymerase chain reaction, RT-PCR), protein kinase C (PKC) activity (isotope incorporation analysis), NF-kappa B activity (electrophoretic mobility shift assay, EMSA), I kappa B-alpha expression (Western blot). Results: After scald, increased expression of TNF-alpha mRNA of PM Phi peaked at 12 PBH. Meanwhile, expression of IL-10 mRNA dropped to the lowest level at 12 PBH. NF-kappa B 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-alpha mRNA. Expression of I kappa B-alpha 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-kappa B inhibitor pyrrolidine dithiocarbamate (PDTC), PKC inhibitor H-7 and PNS, both TNF-alpha mRNA expression and NF-kappa B activity decreased significantly. Conclusions: These results indicate that abnormal expression of TNF-alpha mRNA of macrophages might be regulated by PKC-NF-kappa B signaling following severe burn. PNS might play an anti-inflammatory effect by inhibiting NF-kappa B activity and TNF-alpha mRNA expression. (c) 2006 Elsevier Ltd and ISBI. All rights reserved.
引用
收藏
页码:846 / 852
页数:7
相关论文
共 41 条
[1]  
Abraham E, 1996, New Horiz, V4, P184
[2]  
ALLGOWER M, 1995, BURNS S1, V21, P1
[3]   A RAPID MICROPREPARATION TECHNIQUE FOR EXTRACTION OF DNA-BINDING PROTEINS FROM LIMITING NUMBERS OF MAMMALIAN-CELLS [J].
ANDREWS, NC ;
FALLER, DV .
NUCLEIC ACIDS RESEARCH, 1991, 19 (09) :2499-2499
[4]   The NF-kappa B and I kappa B proteins: New discoveries and insights [J].
Baldwin, AS .
ANNUAL REVIEW OF IMMUNOLOGY, 1996, 14 :649-683
[5]   Systemic inflammatory response syndrome (SIRS), multiple organ dysfunction syndrome (MODS), multiple organ failure (MOF): Are we winning the battle? [J].
Baue, AE ;
Durham, R ;
Faist, E .
SHOCK, 1998, 10 (02) :79-89
[6]   CHARACTERIZATION OF PROTEIN KINASE-C ACTIVITY IN INTERFERON GAMMA TREATED MURINE PERITONEAL-MACROPHAGES [J].
BECTON, DL ;
ADAMS, DO ;
HAMILTON, TA .
JOURNAL OF CELLULAR PHYSIOLOGY, 1985, 125 (03) :485-491
[7]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[8]   Role of oxidants in NF-κB activation and TNF-α gene transcription induced by hypoxia and endotoxin [J].
Chandel, NS ;
Trzyna, WC ;
McClintock, DS ;
Schumacker, PT .
JOURNAL OF IMMUNOLOGY, 2000, 165 (02) :1013-1021
[9]   INTERLEUKIN-10 (IL-10) INHIBITS THE INDUCTION OF NITRIC-OXIDE SYNTHASE BY INTERFERON-GAMMA IN MURINE MACROPHAGES [J].
CUNHA, FQ ;
MONCADA, S ;
LIEW, FY .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1992, 182 (03) :1155-1159
[10]   A role for PKCζ in the LPS-induced translocation NF-κB p65 subunit in cultured myometrial cells [J].
Dallot, E ;
Méhats, C ;
Oger, S ;
Leroy, MJ ;
Breuiller-Fouché, M .
BIOCHIMIE, 2005, 87 (06) :513-521