RESUMO
PURPOSE: In this study, we elucidated the effects of berberine, a major alkaloid component contained in medicinal herbs, such as Phellodendri Cortex and Coptidis Rhizoma, on expression of monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) in a human retinal pigment epithelial cell line (ARPE-19) caused by lipopolysaccharide (LPS) stimulation. METHODS: ARPE-19 cells were cultured to confluence. Berberine and LPS were added to the medium. MCP-1 and IL-8 mRNA were measured by real-time polymerase chain reaction. MCP-1 and IL-8 protein concentrations in the media were measured using enzyme-linked immunosorbent assay. RESULTS: After stimulation with LPS, MCP-1 and IL-8 mRNA in ARPE-19 cells reached maximum levels at 3 h, and MCP-1 and IL-8 protein in the culture media reached maximum levels at 24 h. Berberine dose-dependently inhibited MCP-1 and IL-8 mRNA expression of the cells and protein levels in the media stimulated with LPS. CONCLUSIONS: These findings indicate that berberine inhibited the expression of MCP-1 and IL-8 induced by LPS.
Assuntos
Berberina/farmacologia , Quimiocina CCL2/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-8/genética , Degeneração Macular/genética , Epitélio Pigmentado Ocular/metabolismo , RNA/genética , Células Cultivadas , Quimiocina CCL2/biossíntese , Ensaio de Imunoadsorção Enzimática , Humanos , Interleucina-8/biossíntese , Lipopolissacarídeos/farmacologia , Degeneração Macular/metabolismo , Degeneração Macular/patologia , Epitélio Pigmentado Ocular/efeitos dos fármacos , Epitélio Pigmentado Ocular/patologiaRESUMO
In this study we elucidated the effects of berberine, a major alkaloid component contained in medicinal herbs, such as Phellodendri Cortex and Coptidis Rhizoma, on ischemic neuronal damage in mouse organotypic hippocampal slice cultures (OHSCs) caused by oxygen and glucose deprivation (OGD) and N-methyl-D-aspartate (NMDA) -type glutamate receptor stimulation. Hippocampal slices obtained from 7-d-old ICR mice were cultured for 10 d before the experiments. Ischemia-related damage was induced by OGD (5, 15, 45 min) or NMDA (10 microM) treatment, and was evaluated by measuring propidium iodide (PI) uptake. Levels of apoptotic marker proteins, B-cell lymphoma 2 (Bcl-2) and phosphorylated-Bcl-2 (p-Bcl-2), in the OHSCs were measured as indices of biochemical neuronal cell damage by Western blotting. Berberine (5, 25 microM) or the NMDA antagonist MK-801 (25 microM) was added to the medium 30 min before OGD or NMDA treatment. OGD time-dependently increased PI uptake of the OHSCs. Both berberine (5, 25 microM) and MK-801 (25 microM) significantly inhibited PI uptake at 24 h after 45-min OGD treatment and PI uptake in OHSCs exposed to NMDA for 24 h. OGD treatment also significantly increased the level of p-Bcl-2 but not that of Bcl-2 or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in OHSCs. Berberine (5-25 microM) significantly suppressed the OGD-induced increase of p-Bcl-2 level in OHSCs when tissue was exposed to the alkaloid prior to OGD or simultaneously with OGD. These findings suggest that berberine has protective effects against ischemic damage in mouse OHSCs and that the effects are at least partly mediated by suppression of Bcl-2 phosphorylation.
Assuntos
Berberina/química , Berberina/farmacologia , Ácidos Docosa-Hexaenoicos/farmacologia , Hipocampo/patologia , Neurônios/efeitos dos fármacos , Animais , Berberina/uso terapêutico , Morte Celular/efeitos dos fármacos , Ácidos Docosa-Hexaenoicos/uso terapêutico , Agonistas de Aminoácidos Excitatórios/farmacologia , Glucose/deficiência , Hipocampo/efeitos dos fármacos , Hipocampo/fisiopatologia , Hipóxia/complicações , Hipóxia/tratamento farmacológico , Isquemia/complicações , Isquemia/patologia , Isoxazóis/farmacologia , Isoxazóis/uso terapêutico , Camundongos , Camundongos Endogâmicos ICR , N-Metilaspartato/farmacologia , Técnicas de Cultura de Órgãos , Fosforilação , Propídio , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Sulfonamidas/farmacologia , Sulfonamidas/uso terapêutico , Fatores de TempoRESUMO
PURPOSE: To examine the effects of berberine, an alkaloid isolated from some medicinal herbs, on the disruption of the barrier function in a human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1beta (IL-1beta). METHODS: ARPE-19 cells were cultured to confluence. Berberine and IL-1beta were added to the medium. Barrier functions were evaluated by measuring transepithelial electrical resistance (TER) and the permeability to horseradish peroxidase (HRP) and sodium fluorescein (SF). RESULTS: Berberine dose-dependently inhibited decreased TER and increased the permeability to HRP and SF in the cells stimulated with IL-1beta. CONCLUSIONS: Berberine dose-dependently inhibited the disruption of the barrier function in the ARPE-19 cell line induced by IL-1beta.
Assuntos
Berberina/farmacologia , Barreira Hematorretiniana/fisiologia , Epitélio Pigmentado Ocular/efeitos dos fármacos , Transporte Biológico/efeitos dos fármacos , Linhagem Celular , Permeabilidade da Membrana Celular , Relação Dose-Resposta a Droga , Impedância Elétrica , Eletrofisiologia , Fluoresceína/metabolismo , Peroxidase do Rábano Silvestre/metabolismo , Humanos , Interleucina-1beta/farmacologia , Epitélio Pigmentado Ocular/metabolismoRESUMO
PURPOSE: The aims of this study were to examine the in vivo effects of berberine, an alkaloid isolated from some medicinal herbs, on monocyte chemotactic protein-1 (MCP-1) and cytokine-induced neutrophil chemoattractant-1 (CINC-1) expression in rat lipopolysaccharide (LPS)-induced uveitis. METHODS: LPS was injected intraperitoneally. Berberine was orally administered. MCP-1 mRNA and CINC-1 mRNA were measured by semiquantitative reverse-transcription polymerase chain reaction and real-time polymerase chain reaction. MCP-1 and CINC-1 protein concentration in the aqueous humor were measured by enzyme-linked immunosorbent assay. Histopathologic study was performed in the anterior ocular segments. RESULTS: Berberine dose-dependently inhibited LPS-induced MCP-1 mRNA and CINC-1 mRNA expression of the iris-ciliary body. The alkaloid inhibited chemokines, protein and cell levels in the aqueous humor in rats stimulated with LPS. On histopathologic study, the inflammatory cell infiltration was diminished by the berberine treatment. CONCLUSIONS: These findings indicate that berberine dose-dependently inhibited the expression of MCP-1 and CINC-1 induced by LPS and diminished the anterior uveitis.
Assuntos
Berberina/uso terapêutico , Quimiocina CCL2/genética , Quimiocinas CXC/genética , Expressão Gênica/efeitos dos fármacos , RNA Mensageiro/metabolismo , Uveíte Anterior/tratamento farmacológico , Animais , Quimiocina CXCL1 , Corpo Ciliar/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Iris/metabolismo , Lipopolissacarídeos/toxicidade , Masculino , Reação em Cadeia da Polimerase , Ratos , Ratos Wistar , Uveíte Anterior/induzido quimicamente , Uveíte Anterior/metabolismoRESUMO
We examined the levels of vitreous chemokines and Sho (Zheng in Chinese) of Chinese-Korean-Japanese medicine in diabetic patients. Patients undergoing vitrectomy were classified into Group 1 (no diabetic retinopathy), Group 2 (diabetic retinopathy with no or a few new vessels), and Group 3 (diabetic retinopathy with many new vessels). The levels of IL-8, MCP-1, MIP-1alpha, MIP-1beta, and RANTES in the vitreous fluid were measured using cytometric bead array method. Sho was determined by the standard diagnostic method of Chinese-Korean-Japanese medicine. Vitreous levels of IL-8 and MCP-1 in Groups 2 and 3 were higher than those in Group 1. MIP-1alpha, MIP-1beta, and RANTES levels in Groups 2 and 3 were almost the same as those in Group 1. The percentage of patients with Keishibukuryo-gan (Guizhifuling-wan in Chinese) sho in Group 3 was higher than that in Group 1. In conclusion, vitreous levels of IL-8 and MCP-1 were high in patients with diabetic vitreoretinopathy. Keishibukuryo-gan sho may be associated with diabetic vitreoretinopathy.
Assuntos
Quimiocinas/metabolismo , Retinopatia Diabética/metabolismo , Vitreorretinopatia Proliferativa/metabolismo , Corpo Vítreo/metabolismo , Idoso , Quimiocina CCL2/metabolismo , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CCL5/metabolismo , Diabetes Mellitus Tipo 2/complicações , Retinopatia Diabética/diagnóstico , Retinopatia Diabética/etiologia , Feminino , Humanos , Interleucina-8/metabolismo , Proteínas Inflamatórias de Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Vitrectomia , Vitreorretinopatia Proliferativa/diagnóstico , Vitreorretinopatia Proliferativa/etiologia , Corpo Vítreo/patologia , Corpo Vítreo/cirurgiaRESUMO
PURPOSE: The aims of this study were to examine the effects of berberine, an alkaloid isolated from some medicinal herbs, on interleukin 8 (IL-8) and monocyte chemotactic protein 1 (MCP-1) expression in a human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin 1beta (IL-1beta) or tumor necrosis factor alpha (TNF-alpha). METHODS: ARPE-19 cells were cultured to confluence. Berberine and IL-1beta or TNF-alpha were added to the medium. IL-8 mRNA and MCP-1 mRNA were measured by semiquantitative reverse-transcription polymerase chain reaction and real-time polymerase chain reaction. IL-8 and MCP-1 protein concentrations in the media were measured using enzyme-linked immunosorbent assay. RESULTS: Berberine dose-dependently inhibited IL-8 mRNA and MCP-1 mRNA expression of the cells and protein levels in the media stimulated with IL-1beta or TNF-alpha. CONCLUSION: These findings indicate that berberine dose-dependently inhibited the expression of IL-8 and MCP-1 induced by IL-1beta or TNF-alpha.