Attenuation of inflammatory-mediated neurotoxicity by Saururus chinensis extract in LPS-induced BV-2 microglia cells via regulation of NF-κB signaling and anti-oxidant properties

BMC Complementary and Alternative Medicine

Dong-Kug Choi¹  Ji-Hong Lim²  Pyo-Jam Park¹  Jin-Woo Hwang¹  Shin-Young Park¹  Sushruta Koppula¹  Byung-Wook Kim¹ 

[1] Department of Biotechnology, Konkuk University, Chungju, Korea;Department of Biomedical chemistry, Konkuk University, Chungju, Korea

关键词: Neurodegenerative disease;    NF-κB;    LPS;    Quercetin;    Saururus chinensis;    Microglia;   

Others  :  1084636
DOI  :  10.1186/1472-6882-14-502

 received in 2014-03-13, accepted in 2014-09-19,  发布年份 2014

【 摘 要 】

Background

A Saururus chinensis Baill (SC) has been used by Native Americans, early colonists and practitioners of Korean traditional medicine for treating several diseases including cancer, rheumatoid arthritis and edema. The objective of this study was to evaluate the effects of SC extract in lipopolysaccharide (LPS)-stimulated neuroinflammatory responses in BV-2 microglial cells.

Methods

The effects of SC on the LPS–induced neuroinflammatory responses in BV-2 microglial cells were assessed by Western blotting, RT-PCR and immunofluorescence labeling techniques. DPPH and alkyl radical scavenging assay was performed to evaluate the anti-oxidant effects. Comparisons between groups were analyzed using one-way analysis of variance followed by Dunnett’s multiple comparisons test using GraphPad Prism V5.01 software.

Results

Pre-treatment with SC extract (1, 5 and 10 μg/mL) significantly (p < 0.001 at 10 μg/mL) and concentration dependently inhibited LPS-induced production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2) and suppressed the inflammatory cytokine levels such as tumor necrosis factor-alpha and interleukin (IL)-6 in BV-2 microglial cells (p < 0.001 at 10 μg/mL). Further, SC suppressed the nuclear factor-kappa B (NF-κB) activation by blocking the degradation of IκB-α. SC also exhibited profound anti-oxidant effects by scavenging 1, 1-diphenyl-2-picrylhydrazyl (DPPH) (IC50: 0.055 mg/mL) and alkyl radicals (IC50: 0.349 mg/mL). High performance liquid chromatography finger printing analysis of SC revealed quercetin (QCT) as one of the major constituents compared with reference standard. QCT also inhibited the excessive release of NO, and inhibited the increased expressional levels of IL-6, iNOS and COX-2 in LPS-stimulated BV-2 cells.

Conclusions

Our results indicated that SC inhibited the LPS-stimulated neuroinflammatory responses in BV-2 microglia via regulation of NF-κB signaling. The antioxidant active constituents of SC might be partly involved in delivering such effects. Based on the traditional claims and our present results SC can be potentially used in treating inflammatory-mediated neurodegenerative diseases.

【 授权许可】

   
2014 Kim et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Kim YS, Joh TH: Microglia, major player in the brain inflammation: their roles in the pathogenesis of Parkinson’s disease. Exp Mol Med 2006, 38(4):333-347.
• [2]Koning N, Bo L, Hoek RM, Huitinga I: Downregulation of macrophage inhibitory molecules in multiple sclerosis lesions. Ann Neurol 2007, 62(5):504-514.
• [3]Krause DL, Muller N: Neuroinflammation, microglia and implications for anti-inflammatory treatment in Alzheimer’s disease. Int J Alzheimers Dis 2010, 2010:732806.
• [4]Moller T: Neuroinflammation in Huntington’s disease. J Neural Transm 2010, 117(8):1001-1008.
• [5]Graeber MB, Streit WJ: Microglia: biology and pathology. Acta Neuropathol 2010, 119(1):89-105.
• [6]Chung BS, Shin MG: Dictionary of Korean Folk Medicine. Seoul: Young Lim Publishing Co., Ltd.; 1990.
• [7]Quan Z, Lee YJ, Yang JH, Lu Y, Li Y, Lee YK, Jin M, Kim JY, Choi JH, Son JK, Chang HW: Ethanol extracts of Saururus chinensis suppress ovalbumin-sensitization airway inflammation. J Ethnopharmacol 2010, 132(1):143-149.
• [8]Lee E, Haa K, Yook JM, Jin MH, Seo CS, Son KH, Kim HP, Bae KH, Kang SS, Son JK, Chang HW: Anti-asthmatic activity of an ethanol extract from Saururus chinensis. Biol Pharm Bull 2006, 29(2):211-215.
• [9]Jung MH, Song MC, Bae K, Kim HS, Kim SH, Sung SH, Ye SK, Lee KH, Yun YP, Kim TJ: Sauchinone attenuates oxidative stress-induced skeletal muscle myoblast damage through the down-regulation of ceramide. Biol Pharm Bull 2011, 34(4):575-579.
• [10]Cho HY, Cho CW, Song YS: Antioxidative and anti-inflammatory effects of Saururus chinensis methanol extract in RAW 264.7 macrophages. J Med Food 2005, 8(2):190-197.
• [11]Han HJ, Li M, Son JK, Seo CS, Song SW, Kwak SH, Bae HB: Sauchinone, a lignan from Saururus chinensis, attenuates neutrophil pro-inflammatory activity and acute lung injury. Int Immunopharmacol 2013, 17(2):471-477.
• [12]Yoo HJ, Kang HJ, Jung HJ, Kim K, Lim CJ, Park EH: Anti-inflammatory, anti-angiogenic and anti-nociceptive activities of Saururus chinensis extract. J Ethnopharmacol 2008, 120(2):282-286.
• [13]Choi MS, Kim EC, Lee HS, Kim SK, Choi HM, Park JH, Han JB, An HJ, Um JY, Kim HM, Han AR, Hong MC, Bae H, Min BI: Inhibitory effects of Saururus chinensis (LOUR.) BAILL on the development of atopic dermatitis-like skin lesions in NC/Nga mice. Biol Pharm Bull 2008, 31(1):51-56.
• [14]Lee YJ, Kim J, Yi JM, Oh SM, Kim NS, Kim H, Oh DS, Bang OS, Lee J: Anti-proliferative neolignans from Saururus chinensis against human cancer cell lines. Biol Pharm Bull 2012, 35(8):1361-1366.
• [15]Sung SH, Kim YC: Hepatoprotective diastereomeric lignans from Saururus chinensis herbs. J Nat Prod 2000, 63(7):1019-1021.
• [16]Kim BW, Koppula S, Hong SS, Jeon SB, Kwon JH, Hwang BY, Park EJ, Choi DK: Regulation of microglia activity by glaucocalyxin-A: attenuation of lipopolysaccharide-stimulated neuroinflammation through NF-kappaB and p38 MAPK signaling pathways. PLoS One 2013, 8(2):e55792.
• [17]Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR: Analysis of nitrate, nitrite, and [15 N]nitrate in biological fluids. Anal Biochem 1982, 126(1):131-138.
• [18]Kim BW, Koppula S, Kim JW, Lim HW, Hwang JW, Kim IS, Park PJ, Choi DK: Modulation of LPS-stimulated neuroinflammation in BV-2 microglia by Gastrodia elata: 4-hydroxybenzyl alcohol is the bioactive candidate. J Ethnopharmacol 2012, 139(2):549-557.
• [19]Nanjo F, Goto K, Seto R, Suzuki M, Sakai M, Hara Y: Scavenging effects of tea catechins and their derivatives on 1,1-diphenyl-2-picrylhydrazyl radical. Free Radic Biol Med 1996, 21(6):895-902.
• [20]Wang W, Ji P, Dow KE: Corticotropin-releasing hormone induces proliferation and TNF-alpha release in cultured rat microglia via MAP kinase signalling pathways. J Neurochem 2003, 84(1):189-195.
• [21]Hofmann U, Heuer S, Meder K, Boehler J, Lange V, Quaschning T, Ertl G, Bonz A: The proinflammatory cytokines TNF-alpha and IL-1 beta impair economy of contraction in human myocardium. Cytokine 2007, 39(3):157-162.
• [22]Siebenlist U, Franzoso G, Brown K: Structure, regulation and function of NF-kappa B. Annu Rev Cell Biol 1994, 10:405-455.
• [23]Nomura Y: NF-kappaB activation and IkappaB alpha dynamism involved in iNOS and chemokine induction in astroglial cells. Life Sci 2001, 68(15):1695-1701.
• [24]Li B, Lee DS, Choi HG, Kim KS, Kang DG, Lee HS, Jeong GS, Kim YC: Sauchinone suppresses pro-inflammatory mediators by inducing heme oxygenase-1 in RAW264.7 macrophages. Biol Pharm Bull 2011, 34(10):1566-1571.
• [25]Lee AK, Sung SH, Kim YC, Kim SG: Inhibition of lipopolysaccharide-inducible nitric oxide synthase, TNF-alpha and COX-2 expression by sauchinone effects on I-kappaBalpha phosphorylation, C/EBP and AP-1 activation. Br J Pharmacol 2003, 139(1):11-20.
• [26]Jang EY, Yang CH, Han MH, Choi YH, Hwang M: Sauchinone suppresses lipopolysaccharide-induced inflammatory responses through Akt signaling in BV2 cells. Int Immunopharmacol 2012, 14(2):188-194.
• [27]Lu Y, Li Y, Seo CS, Murakami M, Son JK, Chang HW: Saucerneol D inhibits eicosanoid generation and degranulation through suppression of Syk kinase in mast cells. Food Chem Toxicol 2012, 50(12):4382-4388.
• [28]Lu Y, Suh SJ, Kwak CH, Kwon KM, Seo CS, Li Y, Jin Y, Li X, Hwang SL, Kwon O, Chang YC, Park YG, Park SS, Son JK, Kim CH, Chang HW: Saucerneol F, a new lignan, inhibits iNOS expression via MAPKs, NF-kappaB and AP-1 inactivation in LPS-induced RAW264.7 cells. Int Immunopharmacol 2012, 12(1):175-181.
• [29]Vlad SC, Miller DR, Kowall NW, Felson DT: Protective effects of NSAIDs on the development of Alzheimer disease. Neurology 2008, 70(19):1672-1677.
• [30]Hernan MA, Logroscino G, Garcia Rodriguez LA: Nonsteroidal anti-inflammatory drugs and the incidence of Parkinson disease. Neurology 2006, 66(7):1097-1099.
• [31]Farooqui AA, Horrocks LA, Farooqui T: Modulation of inflammation in brain: a matter of fat. J Neurochem 2007, 101(3):577-599.
• [32]Bauer MK, Lieb K, Schulze-Osthoff K, Berger M, Gebicke-Haerter PJ, Bauer J, Fiebich BL: Expression and regulation of cyclooxygenase-2 in rat microglia. Eur J Biochem 1997, 243(3):726-731.
• [33]Lau LM, Wolter JK, Lau JT, Cheng LS, Smith KM, Hansford LM, Zhang L, Baruchel S, Robinson F, Irwin MS: Cyclooxygenase inhibitors differentially modulate p73 isoforms in neuroblastoma. Oncogene 2009, 28(19):2024-2033.
• [34]Minghetti L, Polazzi E, Nicolini A, Greco A, Levi G: Possible role of microglial prostanoids and free radicals in neuroprotection and neurodegeneration. Adv Exp Med Biol 1999, 468:109-119.
• [35]Teismann P, Vila M, Choi DK, Tieu K, Wu DC, Jackson-Lewis V, Przedborski S: COX-2 and neurodegeneration in Parkinson’s disease. Ann N Y Acad Sci 2003, 991:272-277.
• [36]Hunot S, Hartmann A, Hirsch EC: The inflammatory response in the Parkinson brain. Clinical Neuroscience Research 2001, 1(6):434-443.
• [37]Knott C, Stern G, Wilkin GP: Inflammatory regulators in Parkinson’s disease: iNOS, lipocortin-1, and cyclooxygenases-1 and -2. Mol Cell Neurosci 2000, 16(6):724-739.
• [38]González-Scarano FBG: Microglia as mediators of inflammatory and degenerative diseases. Annu Rev Neurosci 1999, 22:219-240.
• [39]Blasi E, Barluzzi R, Bocchini V, Mazzolla R, Bistoni F: Immortalization of murine microglial cells by a v-raf/v-myc carrying retrovirus. J Neuroimmunol 1990, 27(2–3):229-237.
• [40]Baeuerle PA, Henkel T: Function and activation of NF-kappa B in the immune system. Annu Rev Immunol 1994, 12:141-179.
• [41]Baldwin AS: The transcription factor NF-kappa B and human disease. Journal of Clinical Investigation 2001, 107(1):3-6.
• [42]Delhase M, Li NX, Karin M: Signalling pathways - Kinase regulation in inflammatory response. Nature 2000, 406(6794):367-368.
• [43]Karin M, Ben-Neriah Y: Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. Annu Rev Immunol 2000, 18:621-663.
• [44]Hemmer K, Fransen L, Vanderstichele H, Vanmechelen E, Heuschling P: An in vitro model for the study of microglia-induced neurodegeneration: involvement of nitric oxide and tumor necrosis factor-alpha. Neurochem Int 2001, 38(7):557-565.
• [45]Reis K, Halldin J, Fernaeus S, Pettersson C, Land T: NADPH oxidase inhibitor diphenyliodonium abolishes lipopolysaccharide-induced down-regulation of transferrin receptor expression in N2a and BV-2 cells. Journal of Neuroscience Research 2006, 84(5):1047-1052.
• [46]Bureau G, Longpre F, Martinoli MG: Resveratrol and quercetin, two natural polyphenols, reduce apoptotic neuronal cell death induced by neuroinflammation. J Neurosci Res 2008, 86(2):403-410.
• [47]Casado R, Landa A, Calvo J, Garcia-Mina JM, Marston A, Hostettmann K, Calvo MI: Anti-inflammatory, antioxidant and antifungal activity of Chuquiraga spinosa. Pharm Biol 2011, 49(6):620-626.
• [48]Geraghty DP, Ahuja KD, Pittaway J, Shing C, Jacobson GA, Jager N, Jurkovic S, Narkowicz C, Saunders CI, Ball M, Pinkard A, Vennavaram RR, Adams MJ: In vitro antioxidant, antiplatelet and anti-inflammatory activity of Carpobrotus rossii (pigface) extract. J Ethnopharmacol 2011, 134(1):97-103.
• [49]Jung JY, Lee KY, Lee MY, Jung D, Cho ES, Son HY: Antioxidant and antiasthmatic effects of saucerneol D in a mouse model of airway inflammation. Int Immunopharmacol 2011, 11(6):698-705.
• [50]Li B, Lee YJ, Kim YC, Yoon JJ, Lee SM, Lee YP, Kang DG, Lee HS: Sauchinone from Saururus chinensis protects vascular inflammation by heme oxygenase-1 induction in human umbilical vein endothelial cells. Phytomedicine 2014, 21(2):101-108.
• [51]Park HC, Bae HB, Jeong CW, Lee SH, Jeung HJ, Kwak SH: Effect of manassantin B, a lignan isolated from Saururus chinensis, on lipopolysaccharide-induced interleukin-1beta in RAW 264.7 cells. Korean J Anesthesiol 2012, 62(2):161-165.
• [52]Caddeo C, Diez-Sales O, Pons R, Fernandez-Busquets X, Fadda AM, Manconi M: Topical Anti-Inflammatory Potential of Quercetin in Lipid-Based Nanosystems: In Vivo and In Vitro Evaluation. Pharm Res 2014, 31(4):959-968.
• [53]Woo KW, Moon E, Park SY, Kim SY, Lee KR: Flavonoid glycosides from the leaves of Allium victorialis var. platyphyllum and their anti-neuroinflammatory effects. Bioorg Med Chem Lett 2012, 22(24):7465-7470.
• [54]Bournival J, Plouffe M, Renaud J, Provencher C, Martinoli MG: Quercetin and Sesamin Protect Dopaminergic Cells from MPP+-Induced Neuroinflammation in a Microglial (N9)-Neuronal (PC12) Coculture System. Oxid Med Cell Longev 2012, 2012:921941.
• [55]Kao TK, Ou YC, Raung SL, Lai CY, Liao SL, Chen CJ: Inhibition of nitric oxide production by quercetin in endotoxin/cytokine-stimulated microglia. Life Sci 2010, 86(9–10):315-321.
• [56]Chen JC, Ho FM, Pei-Dawn Lee C, Chen CP, Jeng KC, Hsu HB, Lee ST, Wen Tung W, Lin WW: Inhibition of iNOS gene expression by quercetin is mediated by the inhibition of IkappaB kinase, nuclear factor-kappa B and STAT1, and depends on heme oxygenase-1 induction in mouse BV-2 microglia. Eur J Pharmacol 2005, 521(1–3):9-20.