Dual effect of 3, 4-dihydroxyacetophenone on LPS-induced apoptosis in RAW264. 7 cells by modulating the production of TNF-α

Abstract

To explore the pharmacological effect of 3,4-dihydroxyacetophenone (DHAP) on the apoptosis of RAW264. 7 macrophage cells and the mechanism, RAW264. 7 macrophage cells were treated with 100 or 500 mg/L lipopolysaccharide (LPS), with or without 10⁻⁵ mol/LDHAP for 24 h. Trypan blue dye exclusion assay was used to assess cell viability. Cell apoptosis was morphological studied and flow cytometric assay was used. Tumor necrosis factor-α (TNF-α) level was measured by ELISA methods. IϰB protein was determined by Western blotting. Our results showed that in 100 mg/L LPS-stimulated macrophages, DHAP enhanced the cell apoptosis while in 500 mg/L LPS-stimulated macrophages, DHAP significantly inhibited the cell apoptosis. In both groups, DHAP increased the level of IκB but decreased the level of TNF-α. It is concluded that DHAP has dual effect on the apoptosis of RAW 264. 7 cells treated with different concentrations of LPS. This effect may be due to the inhibition of activation of NF-κB and autocrine production of TNFα. Our study suggests that DHAP may have anti-inflammatory effect on LPS-activated macrophages.

Keywords

3,4-dihydroxyacetophenone / apoptosis / macrophage / TNF-α / NF-κB / inflammation

References

[1] WuX R, LiY, YangD S, et al. . The utero-placental circulation, eugenics and the prevention and treatment of high risk pregnancies. J Tongji Med Univ, 1994, 14(1): 1-1

[2] WuP, HuangY P, YeD Y. Progress in studies on mechanism of promoting circulation and removing blood stasis of 3,4-hydroxyacetophenone. Chin Trad Herb Drugs (Chinese), 2001, 32(3): 277-277

[3] YangD S, WuX R, MaT Y. Effects of 3,4-dihydroxyacetophenone on the biosynthesis of TXA2 and PGI2 in human placental villus and umbilical artery segmentsin vitro. Prostaglandins, 1989, 38(4): 497-497

[4] HuangY P, MaT Y. Effect of 3,4-dihydroxyacetophenone treatment on intrauterine growth retardation. Chin J Obstet Gynecol (Chinese), 1993, 28(6): 333-333

[5] TsuruoT, NaitoM, TomidaA, et al. . Molecular targeting therapy of cancer: drug resistance, apoptosis and survival signal. Cancer Sci, 2003, 94(1): 15-15

[6] RichardM P. Apoptosis as a therapeutic tool in rheumatoid arthritis. Nat Rev Immunol, 2002, 2527-527

[7] HortelanoS, CastrilloA, AlvarezA M, et al. . Contribution of cyclopentenone prostaglandins to the resolution of inflammation through the potentiation of apoptosis in activated macrophages. J Immunol, 2000, 165(11): 6525-6525

[8] JordiX, MònicaC, AnnabelF V, et al. . LPS induces apoptosis in macrophages mostly through the autocrine production of TNF-α. Blood, 2000, 95(12): 3823-3823

[9] MarshC B, WewersM D. The pathogenesis of sepsis. Factors that modulate the response to gram-negative bacterial infection. Clin Chest Med, 1996, 17183-183

[10] KarinM, LinA. NF-kappaB at the crossroads of life and death. Nat Immunol, 2002, 3221-221

[11] SavillJ. Apoptosis in resolution of inflammation. J Leukoc Biol, 1997, 61(4): 375-375

[12] HaslettC. Granulocyte apoptosis and inflammatory disease. Br Med Bull, 1997, 53669-669

[13] vonK A, CallsenD, BruneB. NF-kappaB and AP-1 activation by nitric oxide attenuated apoptotic cell death in RAW 264.7 macrophages. Mol Biol Cell, 1999, 10(2): 361-361

[14] LawrenceT, GilroyD W, Colville-NashP R, et al. . Possible new role for NF-κB in the resolution of inflammation. Nat Med, 2001, 71291-1291

[15] NavarreW W, ZychlinskyA. Pathogen-induced apoptosis of macrophages: a common end for different pathogenic strategies. Cell Microbiol, 2000, 2(4): 265-265

[16] SoniaE, MartaB, CristinaB, et al. . Oxidized phospholipids inhibit cyclooxygenase-2 in human macrophages via nuclear factor-κB/IκB-and ERK2-dependent mechanisms. Cardiovasc Res, 2002, 55406-406