Abstract
Rosmarinic acid (RA) is a natural polyphenolic compound with a well-documented neuroprotective effect mainly associated with its anti-inflammatory and antioxidant activities. Recently, our research group developed and optimized chitosan-coated RA nanoemulsions (RA CNE) intended to be used for nasal delivery as a new potential neuroprotective therapy. In this sense, the present study aimed to evaluate the protective and/or therapeutic potential of RA CNE in inflammation/oxidative stress induced by LPS (1 μg mL−1) in rat astrocyte primary cultures. In summary, pre-treatment with RA CNE before exposure to LPS (protective protocol) reduced significantly the LPS-induced alterations in astrocyte cell viability, proliferation, and cell death by necrosis, which was not observed in therapeutic protocol. RA CNE protective protocol also enhanced anti-oxidative status by ~ 50% by decreasing oxygen reactive species production and nitric oxide levels and preventing total thiol content decrease. Finally, our results demonstrate the protective effect of RA CNE in migratory activation and GFAP expression of reactive astrocytes. Overall, our findings indicate for the first time the RA CNE glioprotective potential, associated with an increase in cell viability and proliferation, a preventive effect on cellular death by necrosis, migratory ability and hypertrophic reactive astrocytes, and the reparation of astrocyte redox state.
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References
Acosta C, Anderson HD, Anderson CM (2017) Astrocyte dysfunction in Alzheimer disease. J Neurosci Res 95:2430–2447. https://doi.org/10.1002/jnr.24075
Aksenov MY, Markesbery WR (2001) Changes in thiol content and expression of glutathione redox system genes in the hippocampus and cerebellum in Alzheimer’s disease. Neurosci Lett 302:141–145. https://doi.org/10.1016/S0304-3940(01)01636-6
Ali S, LeBel C, Bondy S (1992) Reactive oxygen species formation as a biomarker of methylmercury and trimethyltin neurotoxicity. Neurotoxicology 13:637–648
Barnham KJ, Masters CL, Bush AI (2004) Neurodegenerative diseases and oxidative stress. Nat Rev 3:205–214. https://doi.org/10.1038/nrd1330
Bhat AH, Dar KB, Anees S et al (2015) Oxidative stress, mitochondrial dysfunction and neurodegenerative diseases; a mechanistic insight. Biomed Pharmacother 74:101–110. https://doi.org/10.1016/j.biopha.2015.07.025
Bhatt R, Mishra N, Bansal PK (2013) Phytochemical, pharmacological and pharmacokinetics effects of rosmarinic acid. J Pharm Sci Innov 2:28–34. https://doi.org/10.7897/2277-4572.02215
Bhullar KS, Rupasinghe HPV (2013) Polyphenols: multipotent therapeutic agents in neurodegenerative diseases. Oxid Med Cell Longev 2013:1–18. https://doi.org/10.1155/2013/891748
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Braidy N, Matin A, Rossi F et al (2014) Neuroprotective effects of rosmarinic acid on ciguatoxin in primary human neurons. Neurotox Res 25:226–234. https://doi.org/10.1007/s12640-013-9429-9
Casettari L, Illum L (2014) Chitosan in nasal delivery systems for therapeutic drugs. J Control Release 190:189–200. https://doi.org/10.1016/j.jconrel.2014.05.003
Costa P, Sarmento B, Gonçalves S, Romano A (2013) Protective effects of Lavandula viridis L’Hér extracts and rosmarinic acid against H2O2-induced oxidative damage in A172 human astrocyte cell line. Ind Crops Prod 50:361–365. https://doi.org/10.1016/j.indcrop.2013.07.054
de Andrade JM, dos Santos Passos C, Rubio MAK et al (2016) Combining in vitro and in silico approaches to evaluate the multifunctional profile of rosmarinic acid from Blechnum brasiliense on targets related to neurodegeneration. Chem Biol Interact 254:135–145. https://doi.org/10.1016/j.cbi.2016.06.005
Du T, Li L, Song N et al (2010) Rosmarinic acid antagonized 1-methyl-4-phenylpyridinium (MPP +)-induced neurotoxicity in MES23.5 dopaminergic cells. Int J Toxicol 29:625–633. https://doi.org/10.1177/1091581810383705
Dudhani AR, Kosaraju SL (2010) Bioadhesive chitosan nanoparticles: preparation and characterization. Carbohydr Polym 81:243–251. https://doi.org/10.1016/j.carbpol.2010.02.026
Esterbauer H, Cheeseman KH (1990) Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol 186:407–421. https://doi.org/10.1016/0076-6879(90)86134-H
Fachel FNS, Medeiros-Neves B, Dal Prá M et al (2018a) Box-Behnken design optimization of mucoadhesive chitosan-coated nanoemulsions for rosmarinic acid nasal delivery—in vitro studies. Carbohydr Polym 199:572–582. https://doi.org/10.1016/j.carbpol.2018.07.054
Fachel FNS, Nemitz MC, Medeiros-Neves B et al (2018b) A novel, simplified and stability-indicating high-throughput ultra-fast liquid chromatography method for the determination of rosmarinic acid in nanoemulsions, porcine skin and nasal mucosa. J Chromatogr B 1083:233–241. https://doi.org/10.1016/j.jchromb.2018.03.020
Fachel FNS, Schuh RS, Veras KS et al (2019) An overview of the neuroprotective potential of rosmarinic acid and its association with nanotechnology-based delivery systems: a novel approach to treating neurodegenerative disorders. Neurochem Int 122:47–58. https://doi.org/10.1016/j.neuint.2018.11.003
Fallarini S, Miglio G, Paoletti T et al (2009) Clovamide and rosmarinic acid induce neuroprotective effects in in vitro models of neuronal death. Br J Pharmacol 157:1072–1084. https://doi.org/10.1111/j.1476-5381.2009.00213.x
Fernandes C, Oliveira C, Benfeito S et al (2014) Nanotechnology and antioxidant therapy: an emerging approach for neurodegenerative diseases. Curr Med Chem 21:4311–4327. https://doi.org/10.2174/0929867321666140915141836
Fischer R, Maier O (2015) Interrelation of oxidative stress and inflammation in neurodegenerative disease: role of TNF. Oxid Med Cell Longev 2015:1–18
Ghaffari H, Venkataramana M, Ghassam BJ et al (2014) Rosmarinic acid mediated neuroprotective effects against H2O2-induced neuronal cell damage in N2A Cells. Life Sci 113:7–13. https://doi.org/10.1016/j.lfs.2014.07.010
Grassin-Delyle S, Buenestado A, Naline E et al (2012) Intranasal drug delivery: an efficient and non-invasive route for systemic administration—focus on opioids. Pharmacol Ther 134:366–379. https://doi.org/10.1016/j.pharmthera.2012.03.003
Guirado R, Carceller H, Castillo-Gómez E et al (2018) Automated analysis of images for molecular quantification in immunohistochemistry. Heliyon 4:e00669. https://doi.org/10.1016/j.heliyon.2018.e00669
Kim G-D, Park YS, Jin Y-H, Park C-S (2015) Production and applications of rosmarinic acid and structurally related compounds. Appl Microbiol Biotechnol 99:2083–2092. https://doi.org/10.1007/s00253-015-6395-6
Kumar M, Misra A, Babbar AK et al (2008) Intranasal nanoemulsion based brain targeting drug delivery system of risperidone. Int J Pharm 358:285–291. https://doi.org/10.1016/j.ijpharm.2008.03.029
Kuo YC, Rajesh R (2017) Targeted delivery of rosmarinic acid across the blood–brain barrier for neuronal rescue using polyacrylamide-chitosan-poly(lactide-co-glycolide) nanoparticles with surface cross-reacting material 197 and apolipoprotein E. Int J Pharm 528:228–241. https://doi.org/10.1016/j.ijpharm.2017.05.039
Lee HJ, Cho H-S, Park E et al (2008) Rosmarinic acid protects human dopaminergic neuronal cells against hydrogen peroxide-induced apoptosis. Toxicology 250:109–115. https://doi.org/10.1016/j.tox.2008.06.010
Lee AY, Wu TT, Hwang BR et al (2016) The neuro-protective effect of the methanolic extract of Perilla frutescens var. japonica and rosmarinic acid against H2O2—induced oxidative stress in C6 glial cells. Biomol Ther 24:338–345. https://doi.org/10.4062/biomolther.2015.135
Li S, Xie X, Li D et al (2018) Simultaneous determination and tissue distribution studies of four phenolic acids in rat tissue by UFLC–MS/MS after intravenous administration of salvianolic acid for injection. Biomed Chromatogr 32:1–9. https://doi.org/10.1002/bmc.4128
Liddelow SA, Barres BA (2017) Reactive astrocytes: production, function, and therapeutic potential. Immunity 46:957–967. https://doi.org/10.1016/j.immuni.2017.06.006
Liu Z, Zhou T, Ziegler AC et al (2017) Oxidative stress in neurodegenerative diseases: from molecular mechanisms to clinical applications. Oxid Med Cell Longev 2017:1–11. https://doi.org/10.1155/2017/2525967
Lochhead JJ, Thorne RG (2012) Intranasal delivery of biologics to the central nervous system. Adv Drug Deliv Rev 64:614–628. https://doi.org/10.1016/j.addr.2011.11.002
Maragakis NJ, Rothstein JD (2006) Mechanisms of disease: astrocytes in neurodegenerative disease. Nat Clin Pract Neurol 2:679–689
Mistry A, Stolnik S, Illum L (2009) Nanoparticles for direct nose-to-brain delivery of drugs. Int J Pharm 379:146–157. https://doi.org/10.1016/j.ijpharm.2009.06.019
Mujawar N, Ghatage S, Navale S et al (2014) Nasal drug delivery: problem solution and its application. J Curr Pharm Res 4:1231–1245
Noworyta-Sokolowska K, Górska A, Golembiowska K (2013) LPS-induced oxidative stress and inflammatory reaction in the rat striatum. Pharmacol Rep 65:863–869
Pacheco SM, Azambuja JH, de Carvalho TR et al (2018) Glioprotective effects of lingonberry extract against altered cellular viability, acetylcholinesterase activity, and oxidative stress in lipopolysaccharide-treated astrocytes. Cell Mol Neurobiol 38:1107–1121. https://doi.org/10.1007/s10571-018-0581-x
Pérez-Hernández J, Zaldívar-Machorro VJ, Villanueva-Porras D et al (2016) A potential alternative against neurodegenerative diseases. Oxid Med Cell Longev 2016:8378613. https://doi.org/10.1155/2016/8378613
Petersen M, Abdullah Y, Benner J et al (2009) Evolution of rosmarinic acid biosynthesis. Phytochemistry 70(15–16):1663–1679
Prego C, García M, Torres D, Alonso MJ (2005) Transmucosal macromolecular drug delivery. J Control Release 101:151–162. https://doi.org/10.1016/j.jconrel.2004.07.030
Ren P, Jiang H, Li R et al (2009) Rosmarinic acid inhibits 6-OHDA-induced neurotoxicity by anti-oxidation in MES23.5 cells. J Mol Neurosci 39:220–225. https://doi.org/10.1007/s12031-009-9182-y
Shang A-J, Yang Y, Wang H-Y et al (2017) Spinal cord injury effectively ameliorated by neuroprotective effects of rosmarinic acid. Nutr Neurosci 20:172–179. https://doi.org/10.1080/1028415X.2015.1103460
Shinde RL, Jindal AB, Devarajan PV (2011) Microemulsions and nanoemulsions for targeted drug delivery to the brain. Curr Nanosci 7:119–133. https://doi.org/10.2174/157341311794480282
Sofroniew MV (2015) Astrocyte barriers to neurotoxic inflammation. Nat Rev Neurosci 16:249–263. https://doi.org/10.1038/nrn3898
Sofroniew MV, Vinters HV (2010) Astrocytes: biology and pathology. Acta Neuropathol 119:7–35. https://doi.org/10.1007/s00401-009-0619-8
Solanki I, Parihar P, Parihar MS (2016) Neurodegenerative diseases: from available treatments to prospective herbal therapy. Neurochem Int 95:100–108. https://doi.org/10.1016/j.neuint.2015.11.001
Sood S, Jain K, Gowthamarajan K (2014a) Optimization of curcumin nanoemulsion for intranasal delivery using design of experiment and its toxicity assessment. Colloids Surf B 113:330–337. https://doi.org/10.1016/j.colsurfb.2013.09.030
Sood S, Jain K, Gowthamarajan K (2014b) Intranasal therapeutic strategies for management of Alzheimer’s disease. J Drug Target 22:279–294. https://doi.org/10.3109/1061186X.2013.876644
Stuehr DJ, Nathan CF (1989) Nitric oxide. J Exp Med 169:1543–1555
Tezel G, Hernandez MR, Wax MB (2001) In vitro evaluation of reactive astrocyte migration, a component of tissue remodeling in glaucomatous optic nerve head. Glia 34:178–189. https://doi.org/10.1002/glia.1052
Wang X, Chi N, Tang X (2008) Preparation of estradiol chitosan nanoparticles for improving nasal absorption and brain targeting. Eur J Pharm Biopharm 70:735–740. https://doi.org/10.1016/j.ejpb.2008.07.005
Wang J, Li G, Rui T et al (2017) Pharmacokinetics of rosmarinic acid in rats by LC-MS/MS: absolute bioavailability and dose proportionality. RSC Adv 7:9057–9063. https://doi.org/10.1039/C6RA28237G
Zdařilová A, Svobodová A, Šimánek V, Ulrichová J (2009) Prunella vulgaris extract and rosmarinic acid suppress lipopolysaccharide-induced alteration in human gingival fibroblasts. Toxicol Vitr 23:386–392. https://doi.org/10.1016/j.tiv.2008.12.021
Acknowledgements
The authors are gratefully acknowledged to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq—processos 459619/2014-4; 310846/2014-5; 422298/2016-6), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS—processos 16/2551-0000265-7; 16/2551-0000473-0; 17/2551–0001273-9), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES—Finance Code 001) for the financial grants. F.N.S.F. and J.H.A. would like to thank CAPES, CNPq (GD processo 142202/2018-5), and UFCSPA for the doctoral Grant. V.L.B., L.S.K., A.T.H., E.B., and H.F.T. are recipients of CNPq research fellowship.
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FNSF, MDP, JHA, and ME performed the experiments, acquisition of data, analysis and interpretation of data, and wrote the paper. VLB, LSK, ATH, AGB, HFT, and EB made substantial contributions to the conception and design of the study, and critically revised the manuscript. All authors were involved in drafting the manuscript it critically for important intellectual content. All authors gave final approval of the version to be published and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.
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All experiments described in this paper were approved by the Ethics Committee on Animal Use of Federal University of Health Sciences of Porto Alegre, Brazil (protocol number 220/2017).
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Fachel, F.N.S., Dal Prá, M., Azambuja, J.H. et al. Glioprotective Effect of Chitosan-Coated Rosmarinic Acid Nanoemulsions Against Lipopolysaccharide-Induced Inflammation and Oxidative Stress in Rat Astrocyte Primary Cultures. Cell Mol Neurobiol 40, 123–139 (2020). https://doi.org/10.1007/s10571-019-00727-y
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DOI: https://doi.org/10.1007/s10571-019-00727-y