Glioprotective Effect of Chitosan-Coated Rosmarinic Acid Nanoemulsions Against Lipopolysaccharide-Induced Inflammation and Oxidative Stress in Rat Astrocyte Primary Cultures
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.
KeywordsRosmarinic acid Chitosan-coated nanoemulsions Lipopolysaccharide Astrocytes Glioprotective effect
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.
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.
Compliance with Ethical Standards
Conflicts of interest
The authors declare no conflict of interest.
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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