(+)-Vitisin A Inhibits Influenza A Virus-Induced RANTES Production in A549 Alveolar Epithelial Cells through Interference with Akt and STAT₁ Phosphorylation

Abstract

Airway epithelial cells are the initial sites of influenza virus infection. They participate in the airway inflammatory response by expressing various chemokines such as regulated on activation, normal T cell expressed and secreted (RANTES). In the present investigation, the effects of five stilbenes previously isolated from the roots of Vitis thunbergii on RANTES produced by influenza A virus (H1N1)-infected A549 alveolar epithelial cells were studied. We identified (+)-vitisin A, a tetramer of resveratrol, as a potent agent that inhibits RANTES secretion (EC₅₀: 0.27 μM). However, resveratrol exhibited a much smaller effect (EC₅₀: 28.37 μM). H1N1 infection increased the time-dependent phosphorylation of the transcription factor STAT₁ and of Akt (a downstream effector protein of PI3K). When the PI3K-Akt pathway was blocked by wortmannin, H1N1-stimulated STAT₁ phosphorylation and RANTES production were both abrogated, demonstrating that the PI3K-Akt pathway is necessary for STAT₁ activation and RANTES production in A549 cells. Furthermore, H1N1-stimulated phosphorylation of Akt and STAT₁ were also significantly attenuated by (+)-vitisin A. These results suggested that (+)-vitisin A might be a potent anti-inflammatory agent that inhibits influenza A virus-induced RANTES production by interfering with Akt- and STAT₁-related signal pathways.

Abbreviations

CCL:CC chemokine ligand

ERK:extracellular signal-regulated kinase

IFN-γ:interferon-γ

Jak:Janus kinase

MCP-1:monocyte chemoattractant protein-1

NF-κB:nuclear factor-κB

NP-40:nonylphenoxy(polyethoxy)ethanol

Pl3K-Akt:phosphatidylinositol-3-kinase (Pl3K)-Akt

PMSF:phenylmethylsulfonyl fluoride

RANTES:regulated on activation, normal T cell expressed and secreted

SDS:lauryl sulfate sodium salt

STAT:signal transducer and activator of transcription

Vitis thunbergii Sieb. & Zucc. - Vitis ficifolia Bge. - Vitaceae - influenza virus (H1N1) - A549 cells - RANTES - Akt - STAT₁

References

• 1 Folkerts G, Busse W W, Nijkamp F P, Sorkness R, Gern J E. Virus-induced airway hyperresponsiveness and asthma.  Am J Respir Crit Care Med. 1998;  157 1708-20
• 2 Matsukura S, Kokubu F, Noda H, Tokunaga H, Adachi M. Expression of IL-6, IL-8, and RANTES on human bronchial epithelial cells, NCI-H292, induced by influenza virus A.  J Allergy Clin Immunol. 1996;  98 1080-7
• 3 Matsukura S, Kokubu F, Kubo H, Tomita T, Tokunaga H, Kadokura M. et al . Expression of RANTES by normal airway epithelial cells after influenza virus infection.  Am J Respir Cell Mol Biol. 1998;  18 255-64
• 4 Sprenger H, Meyer R G, Kaufmann A, Bussfeld D, Rischkowsky E, Gemsa D. Selective induction of monocyte and not neutrophil-attracting chemokines after influenza A virus infection.  J Exp Med. 1996;  184 1191-6
• 5 Adachi M, Matsukura S, Tokunaga H, Kokubu F. Expression of cytokines on human bronchial epithelial cells induced by influenza virus A.  Int Arch Allergy Immunol. 1997;  113 307-11
• 6 Mark N K, Leung C Y, Wei X Y, Shen X L, Wong R NS, Leung K N. et al . Inhibition of RANTES expression by indirubin in influenza virus-infected human bronchial epithelial cells.  Biochem Pharmacol. 2004;  67 167-74
• 7 Melchjorsen J, Sørensen L N, Paludan S R. Expression and function of chemokines during viral infections: from molecular mechanisms to in vivo function.  J Leukoc Biol. 2003;  74 331-43
• 8 Schindler C, Darnell Jr J E. Transcriptional response to polypeptide ligands: the JAK-STAT pathway.  Annu Rev Biochem. 1995;  64 621-51
• 9 Casola A, Burger N, Liu T, Jamaluddin M, Brasier A R, Garofalo R P. Oxidant tone regulates RANTES gene expression in airway epithelial cells infected with respiratory syncytial virus. Role in viral-induced interferon regulatory factor activation.  J Biol Chem. 2001;  276 19 715-22
• 10 Darnell J E, Kerr I M, Stark G R. Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins.  Science. 1994;  264 1415-21
• 11 Shin Y K, Liu O, Tikoo S K, Babiuk L A, Zhou Y. Effect of the phosphatidylinositol 3-kinase/Akt pathway on influenza A virus propagation.  J Gen Virol. 2007;  88 942-50
• 12 Bian Z M, Elner S G, Yoshida A, Elner V M. Differential involvement of phosphoinositide 3-kinase/Akt in human RPE MCP-1 and IL-8 expression.  Invest Ophthalmol Visc Sci. 2004;  45 1887-96
• 13 Chiu N Y, Chang K H. The illustrated medicinal plants of Taiwan, Vol. 4. Taipei; SMC Publisher 1995: 147-9
• 14 Li W W, Li B G, Chen Y Z. Flexuosol A, a new tetrastilbene from Vitis flexuosa.  J Nat Prod. 1998;  61 646-7
• 15 Huang Y L, Tsai W J, Shen C C, Chen C C. Resveratrol derivatives from the roots of Vitis thunbergii. .  J Nat Prod. 2005;  68 217-20
• 16 Gao X, Xu Y X, Janakiraman N, Chapman R A, Gautam S C. Immunomodulatory activity of resveratrol: suppression of lymphocyte proliferation, development of cell-mediated cytotoxicity, and cytokine production.  Biochem Pharmacol. 2001;  62 1299-308
• 17 Ferrero M E, Bertelli A AE, Fulgenzi A, Pellegatta F, Corsi M M, Bonfrate M. et al . Activity in vitro of resveratrol on granulocyte and monocyte adhesion to endothelium.  Am J Clin Nutr. 1998;  68 1208-14
• 18 Tao H, Wu C, Zhou Y, Gong M, Zhang X, Iribarren P. et al . The grape component resveratrol interferes with the function of chemoattractant receptors on phagocytic leukocytes.  Cell Mol Immunol. 2004;  1 50-6
• 19 Ko H C, Wei B L, Chiou W F. Dual regulatory effect of plant extracts of Forsythia suspense on RANTES and MCP-1 secretion in influenza A virus-infected human bronchial epithelial cells.  J Ethnopharmacol. 2005;  102 418-23
• 20 Don M J, Liao J F, Lin L L, Chiou W F. Cryptotanshinone inhibits chemotactic migration in macrophages through negative regulation of PI3K signaling pathway.  Br J Pharmacol. 2007;  151 38-46
• 21 Huang K S, Li R L, Wang Y H, Lin M. Three new stilbene trimers from the lianas of Gnetum hainanense.  Planta Med. 2001;  67 61-4
• 22 Huang K S, Zhou S, Lin M, Wang Y H. An isorhapontigenin tetramer and a novel stilbene dimer from Gnetum hainanense.  Planta Med. 2002;  68 916-20
• 23 Huang K S, Lin M, Cheng G F. Anti-inflammatory tetramers of resveratrol from the roots of Vitis amurensis and the conformations of the seven-membrane ring in some oligostilbenes.  Phytochemistry. 2001;  58 357-62
• 24 Garcia-Alonso M, Rimbach G, Rivas-Gonzalo J C, De Pascual-Teresa S. Antioxidant and cellular activities of anthocyanins and their corresponding vitisins A - studies in platelets, monocytes, and human endothelial cells.  J Agric Food Chem. 2004;  52 3378-84
• 25 Huang K S, Lin M, Cheng G F. Anti-inflammatory tetramers of resveratrol from the roots of Vitis amurensis and the conformations of the seven-membrane ring in some oligostilbenes.  Phytochemistry. 2001;  58 357-62
• 26 Chan M CW, Cheung C Y, Chui W H, Tsao S W, Nicholls J M, Chan Y O. et al . Proinflammatory cytokine responses induced by influenza A (H5N1) viruses in primary human alveolar and bronchial epithelial cells.  Respir Res. 2005;  6 135-47
• 27 Veckman V, Österlund P, Fagerlund R, Melen K, Matikainen S, Julkunen I. TNF-α and IFN-α enhance influenza-A-virus-induced chemokine gene expression in human A549 lung epithelial cells.  Virology. 2006;  345 96-104