RESUMO
Age-related macular degeneration (AMD) is the leading cause of low vision and blindness for which there is currently no cure. Increased matrix metalloproteinase-9 (MMP-9) was found in AMD and potently contributes to its pathogenesis. Resident microglia also promote the processes of chronic neuroinflammation, accelerating the progression of AMD. The present study investigates the effects and mechanisms of the natural compound theissenolactone B (LB53), isolated from Theissenia cinerea, on the effects of RPE dysregulation and microglia hyperactivation and its retinal protective ability in a sodium iodate (NaIO3)-induced retinal degeneration model of AMD. The fungal component LB53 significantly reduces MMP-9 gelatinolysis in TNF-α-stimulated human RPE cells (ARPE-19). Similarly, LB53 abolishes MMP-9 protein and mRNA expression in ARPE-19 cells. Moreover, LB53 efficiently suppresses nitric oxide (NO) production, iNOS expression, and intracellular ROS levels in LPS-stimulated TLR 4-activated microglial BV-2 cells. According to signaling studies, LB53 specifically targets canonical NF-κB signaling in both ARPE-19 and BV-2 microglia. In an RPE-BV-2 interaction assay, LB53 ameliorates LPS-activated BV-2 conditioned medium-induced MMP-9 activation and expression in the RPE. In NaIO3-induced AMD mouse model, LB53 restores photoreceptor and bipolar cell dysfunction as assessed by electroretinography (ERG). Additionally, LB53 prevents retinal thinning, primarily the photoreceptor, and reduces retinal blood flow from NaIO3 damage evaluated by optic coherence tomography (OCT) and laser speckle flowgraphy (LSFG), respectively. Our results demonstrate that LB53 exerts neuroprotection in a mouse model of AMD, which can be attributed to its anti-retinal inflammatory effects by impeding RPE-mediated MMP-9 activation and anti-microglia.
Assuntos
Degeneração Macular , Degeneração Retiniana , Camundongos , Animais , Humanos , Metaloproteinase 9 da Matriz/metabolismo , Microglia/metabolismo , Epitélio Pigmentado da Retina , Pigmentos da Retina/efeitos adversos , Pigmentos da Retina/metabolismo , Lipopolissacarídeos/farmacologia , Degeneração Macular/induzido quimicamente , Degeneração Macular/tratamento farmacológico , Degeneração Retiniana/metabolismo , Modelos Animais de DoençasRESUMO
During age-related macular degeneration (AMD), chronic inflammatory processes, possibly fueled by high glucose levels, cause a breakdown of the retinal pigment epithelium (RPE), leading to vision loss. Phloretin, a natural dihydroxychalcone found in apples, targets several anti-inflammatory signaling pathways and effectively inhibits transporter-mediated glucose uptake. It could potentially prevent inflammation and cell death of RPE cells through either direct regulation of inflammatory signaling pathways or through amelioration of high glucose levels. To test this hypothesis, ARPE-19 cells were incubated with or without phloretin for 1 h before exposure to lipopolysaccharide (LPS). Cell viability and the release of pro-inflammatory cytokines interleukin 6 (IL-6), IL-8 and vascular endothelial growth factor (VEGF) were measured. Glucose uptake was studied using isotope uptake studies. The nuclear levels of nuclear factor erythroid 2-related factor 2 (Nrf2) were determined alongside the phosphorylation levels of mitogen-activated protein kinases. Phloretin pretreatment reduced the LPS-induced release of IL-6 and IL-8 as well as VEGF. Phloretin increased intracellular levels of reactive oxygen species and nuclear translocation of Nrf2. It also inhibited glucose uptake into ARPE-19 cells and the phosphorylation of Jun-activated kinase (JNK). Subsequent studies revealed that Nrf2, but not the inhibition of glucose uptake or JNK phosphorylation, was the main pathway of phloretin's anti-inflammatory activities. Phloretin was robustly anti-inflammatory in RPE cells and reduced IL-8 secretion via activation of Nrf2 but the evaluation of its potential in the treatment or prevention of AMD requires further studies.
Assuntos
Degeneração Macular , Fator A de Crescimento do Endotélio Vascular , Humanos , Células Epiteliais/metabolismo , Glucose/metabolismo , Inflamação/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Lipopolissacarídeos/toxicidade , Degeneração Macular/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/metabolismo , Floretina/efeitos adversos , Floretina/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Pigmentos da Retina/efeitos adversos , Pigmentos da Retina/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
We investigated the effects of glutathione trisulfide (GSSSG) on lipopolysaccharide (LPS)-induced inflammatory gene expression in immortalized ARPE-19, and primary human and mouse retinal pigment epithelial (RPE) cells. Sulfane sulfur molecules were significantly increased in GSSSG-treated ARPE-19 cells. GSSSG prevented the LPS-induced upregulation of interleukin (IL)-1ß, IL-6, and C-C motif chemokine ligand 2 (CCL2) in ARPE-19/primary RPE cells. Moreover, GSSSG prevented the activation of the nuclear factor-kappa B p65 subunit, and promoted the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) in LPS-treated ARPE-19 cells. ERK1/2 inhibition prevented the GSSSG-mediated inhibition of LPS-induced IL-6 and CCL2 upregulation. Additionally, ERK1/2 activation prevented the upregulation of these genes in the absence of GSSSG. Knockdown of HMOX1 or NRF2, known as anti-oxidative genes, did not affect the activity of GSSSG in the context of LPS stimulation. These findings suggest that GSSSG attenuates LPS-induced inflammatory gene expression via ERK signaling hyperactivation, independently of the NRF2/HMOX1 pathway.
Assuntos
Lipopolissacarídeos , Epitélio Pigmentado da Retina , Animais , Células Epiteliais/metabolismo , Expressão Gênica , Glutationa/análogos & derivados , Humanos , Inflamação/genética , Inflamação/prevenção & controle , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/farmacologia , Pigmentos da Retina/efeitos adversos , Pigmentos da Retina/metabolismo , EnxofreRESUMO
PURPOSE: Lutein and zeaxanthin are macular pigments with a protective function in the retina. These xanthophylls must be obtained from the diet or added to foods or supplements via easy-to-use, stable formulations. The technique employed to produce these formulations may affect the bioavailability of the xanthophylls. METHODS: Forty-eight healthy volunteers were randomized into this double-blind, cross-over study investigating the plasma kinetics of lutein provided as two different beadlet formulations. Subjects (n = 48) received a single dose of 20 mg of lutein as either a starch-matrix ("SMB", FloraGLO® Lutein 5 %) or as a cross-linked alginate-matrix beadlet ("AMB", Lyc-O-Lutein 20 %) formulation. Plasma concentrations of lutein and zeaxanthin were measured at 0, 1, 3, 6, 9, 12, 14, 24, 26, 28, 32, 36, 48, 72, 168, and 672 h. RESULTS: The mean plasma AUC(0-72h), AUC(0-672h), and C(max) for total lutein and zeaxanthin and their all-E-isomers were significantly increased (p < 0.001) from pre-dose concentrations in response to SMB and AMB. There was no difference in lutein T max between the two test articles. However, by 14 h post-dose, total plasma lutein increased by 7 % with AMB and by 126 % with SMB. Total lutein AUC(0-72h) and AUC(0-672h) were 1.8-fold and 1.3-fold higher, respectively, for SMB compared to AMB. Both formulations were well tolerated by subjects in this study. CONCLUSION: These findings confirm that the bioavailability of lutein and zeaxanthin critically depends on the formulation used and document a superiority of the starch-based over the alginate-based product in this study.
Assuntos
Antioxidantes/administração & dosagem , Suplementos Nutricionais , Luteína/administração & dosagem , Xantofilas/administração & dosagem , Adulto , Alginatos/química , Antioxidantes/efeitos adversos , Antioxidantes/química , Antioxidantes/metabolismo , Estudos Cross-Over , Suplementos Nutricionais/efeitos adversos , Método Duplo-Cego , Feminino , Aditivos Alimentares/química , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Humanos , Cinética , Luteína/efeitos adversos , Luteína/análogos & derivados , Luteína/metabolismo , Masculino , Pessoa de Meia-Idade , Valor Nutritivo , Pigmentos da Retina/administração & dosagem , Pigmentos da Retina/efeitos adversos , Pigmentos da Retina/química , Pigmentos da Retina/metabolismo , Amido/química , Estereoisomerismo , Xantofilas/efeitos adversos , Xantofilas/química , Xantofilas/metabolismo , Adulto Jovem , ZeaxantinasRESUMO
PURPOSE: This study was designed to investigate serum and macular response to, and safety of supplementation with, meso-zeaxanthin (MZ), lutein (L), and zeaxanthin (Z), the carotenoids that constitute macular pigment (MP). METHODS: Forty-four healthy subjects were recruited into this randomized, placebo-controlled, clinical trial. Subjects consumed one tablet per day containing 10.6 mg MZ, 5.9 mg L, and 1.2 mg Z (intervention, I group) or placebo (P group). The spatial profile of MP optical density (MPOD) was measured with customized heterochromatic flicker photometry (cHFP), and serum concentrations of L and Z were quantified by using high performance liquid chromatography (HPLC). Subjects were assessed at baseline and at 3 and 6 months. Clinical pathology analysis was performed at baseline and 6 months. RESULTS: Serum concentrations of L and Z increased significantly in the I group (P = 0.001 and 0.003, respectively) and remained stable in the P group (P > 0.05). There was a significant increase in central MPOD in the I group (0.25°: P = 0.001; 0.5°: P = 0.001), with no significant change in the P group (P > 0.05). Clinical pathology analysis confirmed that all variables remained within the normal reference range, with the exception of total cholesterol and low-density lipoprotein (LDL), which exhibited baseline values outside the accepted normal reference range before supplementation. CONCLUSIONS: Subjects supplemented with MZ, L, and Z exhibited significant increases in serum concentrations of these carotenoids and a subsequent increase in central MPOD. Pathology analysis suggested no adverse clinical implications of consuming these carotenoids. (http://isrctn.org number, ISRCTN60816411).