Role of 5 Saponins in Secretion of Cytokines by PRRSV-induced Endothelial Cells

Abstract

The aim of the research presented here was to investigate the immunoloregulatory effects of 5 saponins: Saikosaponins A (SSA) and D (SSD) from Bupleurum chinese DC (Umbelliferae), Panax Notoginseng Saponin (PNS) and Notoginsenoside R1 from Panax notoginseng (Araliaceae) (SR1), and Anemoside B4 from Pulsatilla chinensis Regel (Ranunculaceae) (AB4). To achieve this, endothelial cells were challenged with 10⁵ TCID₅₀/mL PRRSV for 24 h then treated respectively with 5 saponins at 3 concentrations (1, 5 and 10 μg/mL). The cells were incubated at 37°C in a cell incubator for 24 h. The supernatants were collected and analyzed the levels of interleukin-4 (IL-4), IL-10, IL-2, and γ-interferon (IFN-γ) by ELISA kits. The results revealed that PNS and SR1 inhibited the production of IL-4; PNS, SR1 and SSD inhibited the secretion of IL-10; SSA, SSD and AB4 up-regulated IL-2 expression; SSA and SSD increased the level of IFN-γ. All these changes were significant. Taken together, the data suggested that these 5 saponins might effectively regulate immune responses via changes in the levels of these select cytokines.

Key words

IL-2 - IL-4 - IL-10 - IFNγ - PRRSV - saponins

• References

• 1 Cho JG, Dee SA. Porcine reproductive and respiratory syndrome virus. Theriogenology 2006; 66: 655-662
• 2 Wensvoort G, Terpstra C, Pol JM et al. Mystery swine disease in The Netherlands: the isolation of Lelystad virus. The Veterinary quarterly 1991; 13: 121-130
• 3 Russell P, Atkinson K, Krishnan L. Recurrent reproductive failure due to severe placental villitis of unknown etiology. The Journal of reproductive medicine 1980; 24: 93-98
• 4 Wang X, Liu Z. Prevention and treatment of viral respiratory infections by traditional Chinese herbs. Chinese medical journal 2014; 127: 1344-1350
• 5 Na-Bangchang K, Karbwang J. Traditional Herbal Medicine for the Control of Tropical Diseases. Tropical medicine and health 2014; 42 (2 Suppl) 3-13
• 6 Tundis R, Bonesi M, Deguin B et al. Cytotoxic activity and inhibitory effect on nitric oxide production of triterpene saponins from the roots of Physospermum verticillatum (Waldst & Kit) (Apiaceae). Bioorganic & medicinal chemistry 2009; 17: 4542-4547
• 7 Wong VK, Zhou H, Cheung SS et al. Mechanistic study of saikosaponin-d (Ssd) on suppression of murine T lymphocyte activation. Journal of cellular biochemistry 2009; 107: 303-315
• 8 Chen H, Yin J, Deng Y et al. The protective effects of ginsenoside Rg1 against hypertension target-organ damage in spontaneously hypertensive rats. BMC complementary and alternative medicine 2012; 12: 53
• 9 Sengupta S, Toh SA, Sellers LA et al. Modulating angiogenesis: the yin and the yang in ginseng. Circulation 2004; 110: 1219-1225
• 10 Sun B, Xiao J, Sun XB et al. Notoginsenoside R1 attenuates cardiac dysfunction in endotoxemic mice: an insight into oestrogen receptor activation and PI3K/Akt signalling. British journal of pharmacology 2013; 168: 1758-1770
• 11 Liu M, Zhao X, Xiao L et al. Cytotoxicity of the compounds isolated from Pulsatilla chinensis saponins and apoptosis induced by 23-hydroxybetulinic acid. Pharmaceutical biology 2014; 1-9
• 12 Pate M, Damarla V, Chi DS et al. Endothelial cell biology: role in the inflammatory response. Adv Clin Chem 2010; 52: 109-130
• 13 Hu Y, Chen X, Duan H et al. Pulsatilla decoction and its active ingredients inhibit secretion of NO, ET-1, TNF-alpha, and IL-1 alpha in LPS-induced rat intestinal microvascular endothelial cells. Cell biochemistry and function 2009; 27: 284-288
• 14 Lehle K, Straub RH, Morawietz H et al. Relevance of disease- and organ-specific endothelial cells for in vitro research. Cell Biol Int 2010; 34: 1231-1238
• 15 Wang J, Chen X, Zhou Z et al. The inhibitory effect of 3 beta-Hydroxy-12-oleanen-27-oic acid on growth and motility of human hepatoma HepG2 cells through JNK and Akt signaling pathway. Evid Based Complement Alternat Med 2013; 2013: 685159
• 16 Zhou Y, Bai J, Li Y et al. Suppression of immune responses in pigs by nonstructural protein 1 of porcine reproductive and respiratory syndrome virus. Canadian journal of veterinary research=Revue canadienne de recherche veterinaire 2012; 76: 255-260
• 17 Renukaradhya GJ, Alekseev K, Jung K et al. Porcine reproductive and respiratory syndrome virus-induced immunosuppression exacerbates the inflammatory response to porcine respiratory coronavirus in pigs. Viral immunology 2010; 23: 457-466
• 18 Hocke AC, Ermert M, Althoff A et al. Regulation of interleukin IL-4, IL-13, IL-10, and their downstream components in lipopolysaccharide-exposed rat lungs. Comparison of the constitutive expression between rats and humans. Cytokine 2006; 33: 199-211
• 19 Verma S, Nakaoke R, Dohgu S et al. Release of cytokines by brain endothelial cells: A polarized response to lipopolysaccharide. Brain Behav Immun 2006; 20: 449-455
• 20 Hu G, Xue J, Duan H et al. IFN-gamma induces IFN-alpha and IFN-beta expressions in cultured rat intestinal mucosa microvascular endothelial cells. Immunopharmacol Immunotoxicol 2010; 32: 656-662
• 21 Sprague AH, Khalil RA. Inflammatory cytokines in vascular dysfunction and vascular disease. Biochem Pharmacol 2009; 78: 539-552
• 22 Nelms K, Keegan AD, Zamorano J et al. The IL-4 receptor: signaling mechanisms and biologic functions. Annual review of immunology 1999; 17: 701-738
• 23 Shimoda K, van Deursen J, Sangster MY et al. Lack of IL-4-induced Th2 response and IgE class switching in mice with disrupted Stat6 gene. Nature 1996; 380: 630-633
• 24 Ouyang W, Ranganath SH, Weindel K et al. Inhibition of Th1 development mediated by GATA-3 through an IL-4-independent mechanism. Immunity 1998; 9: 745-755
• 25 Garcia-Nicolas O, Quereda JJ, Gomez-Laguna J et al. Cytokines transcript levels in lung and lymphoid organs during genotype 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection. Veterinary immunology and immunopathology 2014; 160: 26-40
• 26 Charerntantanakul W, Yamkanchoo S, Kasinrerk W. Plasmids expressing porcine interferon gamma up-regulate pro-inflammatory cytokine and co-stimulatory molecule expression which are suppressed by porcine reproductive and respiratory syndrome virus. Veterinary immunology and immunopathology 2013; 153: 107-117
• 27 Muraille E, Leo O. Revisiting the Th1/Th2 paradigm. Scandinavian journal of immunology 1998; 47: 1-9
• 28 Levy Y, Brouet JC. Interleukin-10 prevents spontaneous death of germinal center B cells by induction of the bcl-2 protein. The Journal of clinical investigation 1994; 93: 424-428
• 29 Ning N, Dang X, Bai C et al. Panax notoginsenoside produces neuroprotective effects in rat model of acute spinal cord ischemia-reperfusion injury. Journal of ethnopharmacology 2012; 139: 504-512
• 30 Peng XD, Dai LL, Huang CQ et al. Relationship between anti-fibrotic effect of Panax notoginseng saponins and serum cytokines in rat hepatic fibrosis. Biochemical and biophysical research communications 2009; 388: 31-34
• 31 Kato M, Pu MY, Isobe K et al. Characterization of the immunoregulatory action of saikosaponin-d. Cellular immunology 1994; 159: 15-25
• 32 Kato M, Pu MY, Isobe K et al. Cell type-oriented differential modulatory actions of saikosaponin-d on growth responses and DNA fragmentation of lymphocytes triggered by receptor-mediated and receptor-bypassed pathways. Immunopharmacology 1995; 29: 207-213
• 33 Diaz I, Darwich L, Pappaterra G et al. Immune responses of pigs after experimental infection with a European strain of Porcine reproductive and respiratory syndrome virus. The Journal of general virology 2005; 86 (Pt 7) 1943-1951
• 34 Kienzle N, Buttigieg K, Groves P et al. A clonal culture system demonstrates that IL-4 induces a subpopulation of noncytolytic T cells with low CD8, perforin, and granzyme expression. Journal of immunology 2002; 168: 1672-1681
• 35 Karupiah G. Type 1 and type 2 cytokines in antiviral defense. Vet Immunol Immunopathol 1998; 63: 105-109