Anti-Neuroinflammatory effects of the extract of Achillea fragrantissima

BMC Complementary and Alternative Medicine

Rivka Ofir²  Miriam Rindner¹  Alona Telerman¹  Hilla Erlank¹  Sharon Mordechay¹  Anat Elmann¹ 

[1]Department of Food Quality and Safety, Volcani Center, Agricultural Research Organization, Bet Dagan, 50250, Israel

[2]Dead Sea & Arava Science Center and Department of Microbiology & Immunology, Ben-Gurion University of the Negev, Beer-Sheva, 84105, Israel

关键词: cyclooxygenase-2;    matrix metalloproteinase-9;    nitric oxide;    neuroinflammation;    microglial cells;    Achillea fragrantissima;   

Others  :  1232950
DOI  :  10.1186/1472-6882-11-98

 received in 2011-05-24, accepted in 2011-10-21,  发布年份 2011

【 摘 要 】

Background

The neuroinflammatory process plays a central role in the initiation and progression of neurodegenerative diseases such as Parkinson's and Alzheimer's diseases, and involves the activation of brain microglial cells. During the neuroinflammatory process, microglial cells release proinflammatory mediators such as cytokines, matrix metalloproteinases (MMP), Reactive oxygen species (ROS) and nitric oxide (NO). In the present study, extracts from 66 different desert plants were tested for their effect on lipopolysaccharide (LPS) - induced production of NO by primary microglial cells. The extract of Achillea fragrantissima (Af), which is a desert plant that has been used for many years in traditional medicine for the treatment of various diseases, was the most efficient extract, and was further studied for additional anti-neuroinflammatory effects in these cells.

Methods

In the present study, the ethanolic extract prepared from Af was tested for its anti-inflammatory effects on lipopolysaccharide (LPS)-activated primary cultures of brain microglial cells. The levels of the proinflammatory cytokines interleukin1β (IL-1β) and tumor necrosis factor-α (TNFα) secreted by the cells were determined by reverse transcriptase-PCR and Enzyme-linked immunosorbent assay (ELISA), respectively. NO levels secreted by the activate cells were measured using Griess reagent, ROS levels were measured by 2'7'-dichlorofluorescein diacetate (DCF-DA), MMP-9 activity was measured using gel zymography, and the protein levels of the proinflammatory enzymes cyclooxygenase-2 (COX-2) and induced nitric oxide synthase (iNOS) were measured by Western blot analysis. Cell viability was assessed using Lactate dehydrogenase (LDH) activity in the media conditioned by the cells or by the crystal violet cell staining.

Results

We have found that out of the 66 desert plants tested, the extract of Af was the most efficient extract and inhibited ~70% of the NO produced by the LPS-activated microglial cells, without affecting cell viability. In addition, this extract inhibited the LPS - elicited expression of the proinflammatory mediators IL-1β, TNFα, MMP-9, COX-2 and iNOS in these cells.

Conclusions

Thus, phytochemicals present in the Af extract could be beneficial in preventing/treating neurodegenerative diseases in which neuroinflammation is part of the pathophysiology.

【 授权许可】

   
2011 Elmann et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Halliwell B: Oxidativ stress and neurodegeneration: where are we now? J Neurochem 2006, 97:1634-1658.
• [2]Ischiropoulos H, Beckman JS: Oxidativ stress and nitration in neurodegeneration: Cause, effect, or association? J Clin Invest 2003, 111:163-169.
• [3]Minghetti L: Role of inflammation in neurodegenerative diseases. Curr Opin Neurol 2005, 18:315-321.
• [4]Block ML, Hong JS: Microglia and inflammation-mediated neurodegeneration: multiple triggers with a common mechanism. Prog Neurobiol 2005, 76:77-98.
• [5]Minghetti L: Cyclooxygenase-2 (COX-2) in inflammatory and degenerative brain diseases. J Neuropathol Expl Neurol 2004, 63:901-910.
• [6]Calabrese V, Mancuso C, Calvani M, Rizzarelli E, Butterfield DA, Stella AM: Nitric oxide in the central nervous system: neuroprotection versus neurotoxicity. Nat Rev Neurosci 2007, 8:766-775.
• [7]Saha RN, Pahan K: Regulation of inducible nitric oxide synthase gene in glial cells. Antiox Redox Signaling 2006, 8:929-947.
• [8]Cuzzocrea S, Salvemini D: Molecular mechanisms involved in the reciprocal regulation of cyclooxygenase and nitric oxide synthase enzymes. Kidney Int 2007, 71:290-297.
• [9]Gosselin D, Rivest S: Role of IL-1 and TNF in the brain: Twenty years of progress on a Dr. Jekyll/Mr. Hyde duality of the innate immune system. Brain, Behav Immun 2007, 21:281-289.
• [10]Lim GP, Backstrom JR, Cullen MJ, Miller CA, Atkinson RD, Tokes ZA: Matrix metalloproteinases in the neocortex and spinal cord of amyotrophic lateral sclerosis patients. J Neurochem 1996, 67:251-259.
• [11]Deb S, Gottschall PE: Increased production of matrix metalloproteinases in enriched astrocyte and mixed hippocampal cultures treated with beta-amyloid peptides. J Neurochem 1996, 66:1641-1647.
• [12]Rosenberg GA: Matrix metalloproteinases in neuroinflammation. Glia 2002, 39:279-291.
• [13]Gasche Y, Fujimura M, Morita-Fujimura Y, Copin JC, Kawase M, Massengale J, Chan PH: Early appearance of activated matrix metalloproteinase-9 after focal cerebral ischemia in mice: a possible role in blood-brain barrier dysfunction. J Cereb Blood Flow Metab 1999, 19:1020-1028.
• [14]Morita-Fujimura Y, Fujimura M, Gasche Y, Copin JC, Chan PH: Overexpression of copper and zinc superoxide dismutase in transgenic mice prevents the induction and activation of matrix metalloproteinases after cold injury-induced brain trauma. J Cereb Blood Flow Metab 2000, 20:130-138.
• [15]Montaner J, Alvarez-Sabin J, Molina C, Angles A, Abilleira S, Arenillas J, Gonzalez MA, Monasterio J: Matrix metalloproteinase expression after human cardioembolic stroke: temporal profile and relation to neurological impairment. Stroke 2001, 32:1759-1766.
• [16]Kim GW, Gasche Y, Grzeschik S, Copin JC, Maier CM, Chan PH: Neurodegeneratio in striatum induced by the mitochondrial toxin 3-nitropropionic acid: Role of matrix metalloproteinase-9 in early blood-brain barrier disruption? J Neurosci 2003, 23:8733-8742.
• [17]Meli DN, Christen S, Leib SL: Matrix metalloproteinase-9 in pneumococcal meningitis: Activation via an oxidative pathway. J Infect Dis 2003, 187:1411-1415.
• [18]Leib SL, Clements JM, Lindberg RL, Heimgartner C, Loeffler JM, Pfister LA, Tauber MG, Leppert D: Inhibition of matrix metalloproteinases and tumour necrosis factor alpha converting enzyme as adjuvant therapy in pneumococcal meningitis. Brain 2001, 124:1734-1742.
• [19]Leppert D, Leib SL, Grygar C, Miller KM, Schaad UB, Hollander GA: Matrix metalloproteinase (MMP)-8 and MMP-9 in cerebrospinal fluid during bacterial meningitis: association with blood-brain barrier damage and neurological sequelae. Clin Infect Dis 2000, 31:80-84.
• [20]Leib SL, Leppert D, Clements J, Tauber MG: Matrix metalloproteinases contribute to brain damage in experimental pneumococcal meningitis. Infect Immun 2000, 68:615-620.
• [21]Asahi M, Asahi K, Jung JC, del Zoppo GJ, Fini ME, Lo EH: Role for matrix metalloproteinase 9 after focal cerebral ischemia: effects of gene knockout and enzyme inhibition with BB-94. J Cereb Blood Flow Metab 2000, 20:1681-1689.
• [22]Asahi M, Wang X, Mori T, Sumii T, Jung JC, Moskowitz MA, Fini ME, Lo EH: Effects of matrix metalloproteinase-9 gene knock-out on the proteolysis of blood-brain barrier and white matter components after cerebral ischemia. J Neurosci 2001, 21:7724-7732.
• [23]Wang X, Jung J, Asahi M, Chwang W, Russo L, Moskowitz MA, Dixon CE, Fini ME, Lo EH: Effects of matrix metalloproteinase-9 gene knock-out on morphological and motor outcomes after traumatic brain injury. J Neurosci 2000, 20:7037-7042.
• [24]Dubois B, Masure S, Hurtenbach U, Paemen L, Heremans H, van den Oord J, Sciot R, Meinhardt T, Hammerling G, Opdenakker G, Arnold B: Resistance of young gelatinase B-deficient mice to experimental autoimmune encephalomyelitis and necrotizing tail lesions. J Clin Invest 1999, 104:1507-1515.
• [25]Shabana MM, Mirhom YW, Genenah AA, Aboutabl EA, Amer HA: Study Into Wild Egyptian Plants Of Potential Medicinal Activity. Ninth communication: Hypoglycemic Activity Of Some Selected Plants In Normal Fasting And Alloxanized Rats. Arch Exp Veterinarmedizin 1990, 44:389-394.
• [26]Mustafa EH, Abu Zarga M, Abdalla S: Effects of cirsiliol, a flavone isolated from Achillea fragrantissima, on rat isolated ileum. Gen Pharmacol 1992, 23:555-560.
• [27]Yaniv Z, Dafni A, Friedman J, Palevitch D: Plants used for the treatment of diabetes in Israel. J Ethnopharmacol 1987, 19:145-151.
• [28]Hamdan I, Afifi FU: Studies on the in vitro and in vivo hypoglycemic activities of some medicinal plants used in treatment of diabetes in Jordanian traditional medicine. J Ethnopharmacol 2004, 93:117-121.
• [29]Elmann A, Mordechay S, Rindner M, Ravid U: Anti-neuroinflammatory effects of the essential oil from Pelargonium graveolens in microglial cells. J Func Foods 2009, 2:17-22.
• [30]Kueng W, Silber E, Eppenberger U: Quantification of cells cultured on 96-well plates. Anal Biochem 1989, 182:16-19.
• [31]Bowry VW, Stocker R: Tochoferol-mediated oxidation. The prooxidant effect of vitamin E on the radical-initiated oxidation of human low density lipoproteins. J Am Chem Soc 1993, 115:6029-6044.
• [32]Niki E, Saito M, Yoshikawa Y, Yamamoto Y, Kamiya Y: Oxidation of lipids XII. Inhibition of oxidation of soybean phosphatidylcholine and methyl linoleate in aqueous dispersions by uric acid. Bull Chem Soc Jpn 1986, 59:471-477.
• [33]Thomas MJ, Chen Q, Franklin C, Rudel LL: A comparison of the kinetics of low-density lipoprotein oxidation initiated by copper or by azobis (2-amidinopropane). Free Radic Biol Med 1997, 23:927-935.
• [34]Wolfe KL, Liu RH: Cellular antioxidant activity (CAA) assay for assessing antioxidants, foods, and dietary supplements. J Agric Food Chem 2007, 55:8896-8897.
• [35]Mollace V, Colasanti M, Muscoli C, Lauro G, Iannone M, Rotiroti D, Nistico G: The effect of nitric oxide on cytokine-induced release of PGE2 by human cultured astroglial cells. Br J Pharmacol 1998, 124:742-746.
• [36]Yong VW, Krekoski CA, Forsyth PA, Bell R, Edwards DR: Matrix metalloproteinases and diseases of the CNS. Trends Neurosci 1998, 21:75-80.
• [37]Gu Z, Kaul M, Yan B, Kridel SJ, Cui J, Strongin A, Smith JW, Liddington RC, Lipton SA: S-nitrosylation of matrix metalloproteinases: signaling pathway to neuronal cell death. Science (New York, N.Y.) 2002, 297:1186-1190.
• [38]Wang HH, Hsieh HL, Yang CM: Nitric oxide production by endothelin-1 enhances astrocytic migration via the tyrosine nitration of matrix metalloproteinase-9. J Cell Physiol 2011, 226:2244-2256.
• [39]Salvemini D, Seibert K, Masferrer JL, Settle SL, Currie MG, Needleman P: Nitric Oxide Activates the Cyclooxygenase Pathway in Inflammation. Am J Ther 1995, 2:616-619.
• [40]Marnett LJ, Wright TL, Crews BC, Tannenbaum SR, Morrow JD: Regulation of prostaglandin biosynthesis by nitric oxide is revealed by targeted deletion of inducible nitric-oxide synthase. J Biol Chem 2000, 275:13427-13430.
• [41]Wu CY, Hsieh HL, Jou MJ, Yang CM: Involvement of p42/p44 MAPK, p38 MAPK, JNK and nuclear factor-kappa B in interleukin-1beta-induced matrix metalloproteinase-9 expression in rat brain astrocytes. J Neurochem 2004, 90:1477-1488.
• [42]Liu SF, Malik B: NF-kB activation as a pathological mechanism of septic shock and inflammation. Am J Physiol Lung Cell Mol Physiol 2006, 290:L622-L645.
• [43]Fiebich BL, Butcher RD, Gebicke-Haerter PJ: Protein kinase C-mediated regulation of inducible nitric oxide synthase expression in cultured microglial cells. J Neuroimmunol 1998, 92:170-178.
• [44]Bhat NR, Zhang P, Lee JC, Hogan EL: Extracellular signal-regulated kinase and p38 subgroups of mitogen-activated protein kinases regulate inducible nitric oxide synthase and tumor necrosis factor-alpha gene expression in endotoxin-stimulated primary glial cultures. J Neurosci 1998, 18:1633-1641.
• [45]Sondergaard BC, Schultz N, Madsen SH, Bay-Jensen AC, Kassem M, Karsdal MA: MAPKs are essential upstream signaling pathways in proteolytic cartilage degradation--divergence in pathways leading to aggrecanase and MMP-mediated articular cartilage degradation. Osteoarthritis Cartilage 2010, 18:279-288.
• [46]Auer M, Pfister LA, Leppert D, Tauber MG, Leib SL: Effects of clinically used antioxidants in experimental pneumococcal meningitis. J Infect Dis 2000, 182:347-350.
• [47]Aid S, Bosetti F: Targeting cyclooxygenases-1 and -2 in neuroinflammation: Therapeutic implications. Biochimie 2011, 93:46-51.
• [48]Allan SM, Pinteaux E: The interleukin-1 system: an attractive and viable therapeutic target in neurodegenerative disease. Curr Drug Targets: CNS Neurol Disord 2003, 2:293-302.
• [49]Rothwell NJ, Luheshi GN: Interleukin 1 in the brain: biology, pathology and therapeutic target. Trends neurosci 2000, 23:618-625.