摘要 To investigate whether lactic acid could inhibit the LPS-activation of NF-κB p65 in rat intestinal mucosa microvascular endothelial cells (RIMMVECs), RIMMVECs, cultured in vitro, were pretreated with different concentrations of lactic acid and then exposed to lipopolysaccharide (LPS). Cells and cell culture media were then collected at different time intervals.Production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) was examined at the protein level by enzyme-linked immunosorbent assay. The influence of lactic acid on the LPS-activation of NF-κB was examined at mRNA and protein levels by real-time quantitative PCR and Western blot analysis, respectively. TNF-α and IL-6 protein levels were significantly decreased after pretreatment with lactic acid compared with cells exposed to LPS only. After pretreatment with 7.5, 5.0, and 2.5 μL mL-1 lactic acid, NF-κB mRNA levels were increased by 1.51-, 2.62- and 3.00-fold, respectively, compared with levels in control cells without LPS treatment. Western blot analysis indicated that the level of NF-κB p65 in the lactic acidpretreated group was significantly lower than that in the group treated with LPS only (positive control) and was unchanged compared with the group without LPS treatment (blank control). These results suggest that lactic acid may inhibit LPSactivation of NF-κB, leading to the down-regulation of TNF-α and IL-6.
Abstract To investigate whether lactic acid could inhibit the LPS-activation of NF-κB p65 in rat intestinal mucosa microvascular endothelial cells (RIMMVECs), RIMMVECs, cultured in vitro, were pretreated with different concentrations of lactic acid and then exposed to lipopolysaccharide (LPS). Cells and cell culture media were then collected at different time intervals.Production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) was examined at the protein level by enzyme-linked immunosorbent assay. The influence of lactic acid on the LPS-activation of NF-κB was examined at mRNA and protein levels by real-time quantitative PCR and Western blot analysis, respectively. TNF-α and IL-6 protein levels were significantly decreased after pretreatment with lactic acid compared with cells exposed to LPS only. After pretreatment with 7.5, 5.0, and 2.5 μL mL-1 lactic acid, NF-κB mRNA levels were increased by 1.51-, 2.62- and 3.00-fold, respectively, compared with levels in control cells without LPS treatment. Western blot analysis indicated that the level of NF-κB p65 in the lactic acidpretreated group was significantly lower than that in the group treated with LPS only (positive control) and was unchanged compared with the group without LPS treatment (blank control). These results suggest that lactic acid may inhibit LPSactivation of NF-κB, leading to the down-regulation of TNF-α and IL-6.
LIU Jing, XUE Jiu-zhou, ZHU Zhi-ning, HU Ge, REN Xiao-ming.
2011.
Lactic Acid Inhibits NF-κB Activation by Lipopolysaccharide in Rat Intestinal Mucosa Microvascular Endothelial Cells. Journal of Integrative Agriculture, 10(6): 954-959.
Calzado M A, Bacher S, Schmitz M L. 2007. NF-κB inhibitors for the treatment of inflammatory diseases and cancer.
Current Medicinal Chemistry, 14, 367-376.
Camandola S, Mattson M P. 2007. NF-κB as a therapeutic target in neurodegenerative diseases. Expert Opinion on Therapeutic Targets, 11, 123-132.
Choi J, Enis D R, Koh K P, Shiao S L, Pober J S. 2004. T lymphocyte-endothelial cell interactions. Annual Review of Immunology, 22, 683-689.
Collins T, Read M A, Neish A S, Whitley M Z, Cordle S R, Donald R, Read M A, Hawiger J. 1993. Lipopolysaccharide induces phosphorylation of MAD3 and activation of c-Rel and related NF-κB proteins in human monocytic THP-1 cells. Journal of Biological Chemistry, 268, 11803- 11810.
Feldmann M, Maini R N. 2001. Anti-TNF alpha therapy of rheumatoid arthritis: what have we learned. Annual Review of Immunology, 19, 163-196.
Hindryckx P, de Vos M, Jacques P, Ferdinande L, Peeters H, Olievier K, Bogaert S, Brinkman B, Vandenabeele P, Elewaut D, et al. 2010. Hydroxylase inhibition abrogates TNF-α- induced intestinal epithelial damage by hypoxia-inducible factor-1-dependent repression of FADD. The Journal of Immunology, 185, 6306-6316.
Kaileh M, Sen R. 2010. Role of NF-κB in the anti-inflammatory effects of Tocotrienols. Journal of the American College of Nutrition, 29, 334s-339s.
Karin M. 2006. Nuclear factor-κB in cancer development andprogression. Nature, 441, 431-436.
Kerekes G, Szekanecz Z, Dér H, Sandor Z, Lakos G, Muszbek L, Csipó I, Sipka S, Seres I, Paragh G, et al. 2008. Endothelial dysfunction and atherosclerosis in rheumatoid arthritis: a multiparametric analysis using imaging techniques and laboratory markers of inflammation and autoimmunity. The Journal of Rheumatology, 35, 398-406.
Kishimoto T. 2010. IL-6: from its discovery to clinical applications. International Immunology, 22, 347-352.
Kishimoto T, Ishizaka K. 1971. Regulation of antibody response in vitro. Suppression of secondary response by antiimmunoglobulin heavy chains. The Journal of Immunology, 107, 1567-1570.
O’Sullivan B, Thompson A, Thomas R. 2007. NF-κB as a therapeutic target in autoimmune disease. Expert Opinion on Therapeutic Targets, 11, 111-122.
Pahl H L. 1999. Activators and target genes of Rel’NF-κB transcription factors. Oncogene, 18, 6853-6866.
Senftleben U, Karin M. 2002. The IKK/NF-κB pathway. Critical Care Medicine, 30, S105-111.
Stankova J, D’Orleans-Juste P, Rola-Pleszczynski M. 1996. ET-1 induces IL-6 gene expression in human umbilical vein endothelial cells: synergistic effect of IL-1. American Journal of Physiology - Cell Physiology, 271, C1073-C1078.
Sun D, Lytle C, O´Donnell M E. 1997. IL-6 secreted by astroglial cells regulates Na-K-Cl cotransport in brain microvessel endothelial cells. American Journal of Physiology - Cell Physiology, 272, C1829-C1835.
Tabruyn S P, Mémet S, Avé P, Verhaeghe C, Mayo K H, Struman I, Martial J A, Griffioen A W. 2009. NF-κB activation in endothelial cells is critical for the activity of angiostatic agents. Molecular Cancer Therapeutics, 8, 2645-2654.
Tak P P, Firestein G S. 2001. NF-κB: a key role in inflammatory disseases. The Journal of Clinical Investigation, 107, 7-11. Wulczyn F G, Krappmann D, Scheidereit C. 1996. The NF-κBRel and IκB gene families: mediators of immune response and inflammation. Journal of Molecular Medicine, 74, 749- 769.
Yorek M A, Dunlap J A, Thomas M J, Cammarata P R, Zhou C, Lowe W L. 1998. Effect of TNF-a on SMIT mRNA levels and myo-inositol accumulation in cultured endothelial cells. American Journal of Physiology - Cell Physiology, 274, 58- 71.
Zingarelli B, Sheehan M, Wong H R. 1997. Nuclear factor-κB as a therapeutic expression by PPM-18, a novel antiinflammatory agent, in vitro and in vivo. Biochemical Journal, 328, 363-369.
