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1.
J Exp Clin Cancer Res ; 38(1): 371, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31438982

RESUMO

BACKGROUND: Arachidonic acid (AA) metabolic enzymes including cyclooxygenase-2 (COX-2), microsomal prostaglandin E synthase-1 (mPGES-1) and cytochrome P450 (CYP) 4A11 play important roles in glioma angiogenesis. Thus, there is an urgent need to identify the underlying mechanisms and develop strategies to overcome them. METHODS: A homology model of human CYP4A11 was constructed using SYBYL-X 2.0. Structure-based virtual screening against COX-2, mPGES-1 and CYP4A11was performed using the Surflex-Dock of the SYBYL suite. The candidates were further evaluated their antiangiogenic activities in a zebrafish embryo and rabbit corneal angiogenesis model. Laser doppler analysis was used to measure tumor perfusion. The expression of CD31 and α-SMA was measured by immunofluorescence. Western blot was used to measure the expression of HIF-1, Akt and p-Akt. The gene expression of FGF-2, G-CSF, PDGF, TGF-ß, Tie-2, VEGF, lncRNA NEAT1 and miR-194-5p were determined using qPCR. The production of FGF-2, TGF-ß and VEGF were analyzed using ELISA. Bioinformatic analysis and luciferase reporter assays confirmed the interaction between lncRNA NEAT1 and miR-194-5p. RESULTS: The nearly 36,043 compounds from the Traditional Chinese Medicine (TCM) database were screened against COX-2, mPGES-1 and CYP4A11 3D models, and the 17 top flavonoids were identified. In zebrafish screening, isoliquiritigenin (ISL) exhibited the most potent antiangiogenic activities with the EC50 values of 5.9 µM. Conversely, the antiangiogenic effects of ISL in the zebrafish and rabbit corneal models were partly reversed by 20-hydroxyeicosatetraenoic acid (20-HETE) or prostaglandin E2 (PGE2). ISL normalized glioma vasculature and improved the efficacy of temozolomide therapy in the rat C6 glioma model. Inhibition of COX-2, mPGES-1 and CYP4A by ISL decreased FGF-2, TGF-ß and VEGF production in the C6 and U87 glioma cells with p-Akt downregulation, which was reversed by Akt overexpression. Furthermore, ISL downregulated lncRNA NEAT1 but upregulated miR-194-5p in the U87 glioma cell. Importantly, lncRNA NEAT1 overexpression reversed ISL-mediated increase in miR-194-5p expression, and thereby attenuated FGF-2, TGF-ß and VEGF production. CONCLUSIONS: Reprogramming COX-2, mPGES-1 and CYP4A mediated-AA metabolism in glioma by flavonoid ISL inhibits the angiogenic Akt- FGF-2/TGF-ß/VEGF signaling through ceRNA effect of miR-194-5p and lncRNA NEAT1, and may serve as a novel therapeutic strategy for human glioma.


Assuntos
Chalconas/farmacologia , Ciclo-Oxigenase 2/química , Citocromo P-450 CYP4A/antagonistas & inibidores , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioma/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Prostaglandina-E Sintases/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Animais , Apoptose , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Proliferação de Células , Neovascularização da Córnea/tratamento farmacológico , Neovascularização da Córnea/metabolismo , Neovascularização da Córnea/patologia , Ciclo-Oxigenase 2/metabolismo , Citocromo P-450 CYP4A/metabolismo , Inibidores Enzimáticos/farmacologia , Glioma/irrigação sanguínea , Glioma/metabolismo , Glioma/patologia , Humanos , Masculino , MicroRNAs/genética , Prostaglandina-E Sintases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/genética , Coelhos , Ratos , Ratos Wistar , Células Tumorais Cultivadas , Peixe-Zebra
2.
Exp Eye Res ; 185: 107697, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31228461

RESUMO

Corneal neovascularization (CNV) is associated with different ocular pathologies, including infectious keratitis, trachoma or corneal trauma. Pharmacological treatments based on the topical application of anti-VEGF therapies have been shown to be effective in the treatment and prevention of CNV. The aim of this work was to evaluate the effect of bevacizumab-loaded albumin nanoparticles in a rat model of CNV. Bevacizumab-loaded nanoparticles, either "naked" (B-NP) or coated with PEG 35,000 (B-NP-PEG), were administered once a day in the eyes of animals (10 µL, 4 mg/mL every 24 h) during 7 days. Bevacizumab and dexamethasone were employed as controls and administered at the same dose every 12 h. At the end of the study, the area of the eye affected by neovascularization was about 2-times lower for animals treated with B-NP than with free bevacizumab. In the study, dexamethasone did not demonstrate an inhibitory effect on CNV at the employed dose. All of these results were confirmed by histopathological analysis, which clearly showed that eyes treated with nanoparticles displayed lower levels of fibrosis, inflammation and edema. In summary, the encapsulation of bevacizumab in human serum albumin nanoparticles improved its efficacy in an animal model of CNV.


Assuntos
Inibidores da Angiogênese/uso terapêutico , Bevacizumab/uso terapêutico , Neovascularização da Córnea/tratamento farmacológico , Modelos Animais de Doenças , Portadores de Fármacos/química , Nanopartículas/química , Albumina Sérica Humana/química , Animais , Materiais Revestidos Biocompatíveis , Neovascularização da Córnea/patologia , Masculino , Polietilenoglicóis , Ratos , Ratos Wistar , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores
3.
EBioMedicine ; 44: 542-553, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31126890

RESUMO

BACKGROUND: Corneal neovascularization (angiogenesis and lymphangiogenesis) compromises corneal transparency and transplant survival, however, the molecular mechanisms of corneal host epithelial and stromal cells in neovascularization have not yet been fully elucidated. Furthermore, the contribution and mechanism of corneal host endothelial cells involved in neovascularization are largely unexplored. METHODS: Liquid chromatography-mass spectrometry, immunoblotting, and ELISA were used to screen and identify potential neovascularization-related factors in human full-thickness vascularized corneal tissues. Lipopolysaccharide was used to induce inflammation in three kinds of corneal host cells in vitro, including corneal epithelial, stromal, and endothelial cells. Fungus was used to establish an animal model of corneal neovascularization in vivo. Tube formation and spheroid sprouting assays were used to evaluate the contribution of three kinds of corneal host cells to the degree of neovascularization under various stimuli. Matrix metalloproteinase (MMP)-2, alpha-crystallin A chain (CRYAA), galectin-8, Bcl-2, neuropilin-2, MMP-9 plasmids, and recombinant human fibronectin were used to identify the key proteins of corneal host cells involved in corneal inflammatory neovascularization. FINDINGS: All three kinds of corneal host cells influenced corneal neovascularization to varying degrees. MMP-9 in human corneal epithelial cells, MMP-2, and CRYAA in human corneal stromal cells, and MMP-2 and galectin-8 in human corneal endothelial cells are potential key proteins that participate in corneal inflammatory neovascularization. INTERPRETATION: Our data indicated that both the effects of key proteins and corneal host cells involved should be considered for the treatment of corneal inflammatory neovascularization.


Assuntos
Córnea/citologia , Neovascularização da Córnea/etiologia , Neovascularização da Córnea/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Biomarcadores , Linhagem Celular , Cromatografia Líquida , Córnea/metabolismo , Neovascularização da Córnea/metabolismo , Meios de Cultivo Condicionados , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/metabolismo , Feminino , Expressão Gênica , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Proteoma , Proteômica , Ratos , Células Estromais/metabolismo , Adulto Jovem
4.
Nutrients ; 11(5)2019 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-31137826

RESUMO

Severe corneal inflammation produces opacity or even perforation, scarring, and angiogenesis, resulting in blindness. In this study, we used the cornea to examine the effect of new anti-angiogenic chemopreventive agents. We researched the anti-angiogenic effect of two extracts, methanol (Met) and hexane (Hex), from the seed of Cucurbita argyrosperma, on inflamed corneas. The corneas of Wistar rats were alkali-injured and treated intragastrically for seven successive days. We evaluated: opacity score, corneal neovascularization (CNV) area, re-epithelialization percentage, and histological changes. Also, we assessed the inflammatory (cyclooxigenase-2, nuclear factor-kappaB, and interleukin-1ß) and angiogenic (vascular endothelial growth factor A, VEGF-A; -receptor 1, VEGFR1; and -receptor 2, VEGFR2) markers. Levels of Cox-2, Il-1ß, and Vegf-a mRNA were also determined. After treatment, we observed a reduction in corneal edema, with lower opacity scores and cell infiltration compared to untreated rats. Treatment also accelerated wound healing and decreased the CNV area. The staining of inflammatory and angiogenic factors was significantly decreased and related to a down-expression of Cox-2, Il-1ß, and Vegf. These results suggest that intake of C. argyrosperma seed has the potential to attenuate the angiogenesis secondary to inflammation in corneal chemical damage.


Assuntos
Inibidores da Angiogênese/farmacologia , Anti-Inflamatórios/farmacologia , Queimaduras Químicas/tratamento farmacológico , Córnea/irrigação sanguínea , Córnea/efeitos dos fármacos , Neovascularização da Córnea/tratamento farmacológico , Cucurbita , Queimaduras Oculares/tratamento farmacológico , Extratos Vegetais/farmacologia , Sementes , Inibidores da Angiogênese/isolamento & purificação , Proteínas Angiogênicas/metabolismo , Animais , Anti-Inflamatórios/isolamento & purificação , Queimaduras Químicas/metabolismo , Queimaduras Químicas/patologia , Córnea/metabolismo , Neovascularização da Córnea/metabolismo , Neovascularização da Córnea/patologia , Opacidade da Córnea/tratamento farmacológico , Opacidade da Córnea/metabolismo , Opacidade da Córnea/patologia , Cucurbita/química , Modelos Animais de Doenças , Queimaduras Oculares/metabolismo , Queimaduras Oculares/patologia , Mediadores da Inflamação/metabolismo , Masculino , Extratos Vegetais/isolamento & purificação , Ratos Wistar , Sementes/química , Transdução de Sinais/efeitos dos fármacos , Cicatrização/efeitos dos fármacos
5.
Cutan Ocul Toxicol ; 38(4): 356-359, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31137972

RESUMO

Purpose: To investigate the effect of Hypericum perforatum on corneal alkali burn. Methods: We studied 45 250 g weighing, 4 months old Wistar albino rats. Alkaline burns were performed in the corneas of all experimental animals with 2 mol/L NaOH after general anaesthesia. Rats were divided into five groups according to the subsequent process applied to them: group 1 was the topical Hypericum perforatum group, group 2 was the topical pure olive oil group, group 3 was the oral Hypericum perforatum group, group 4 was the oral pure olive oil group, and group 5 was the control untreated group. Rats were sacrificed under general anaesthesia on the 14 day. The rate of corneal inflammation, neovascularization, fibroblastic activity, and cluster of differentiation 31 (CD31) staining was investigated. Result: There were 45 rats at the beginning of the study. One rat in groups 1, 2, and 3 died during the study; therefore, 42 rats could be evaluated. There were 8 rats in group 1, 8 rats in group 2, 8 rats in group 3, and 9 rats in group 4. We found less corneal neovascularization (CNV), inflammation, and fibroblastic activity in group 1 and group 2 than in the other groups (p ˂ 0.001 for all parameters). CNV, inflammation, fibroblastic activity, and CD31 staining rates were lower in group 1 than in group 2 (p ˂ 0.001 for all parameters). There was no difference between groups 3, 4, and 5 (respectively, p = 0.436, 0.634, and 0.750). Conclusions: We found that both topical Hypericum perforatum oily extract and olive oil have anti-inflammatory, anti-angiogenic, and anti-fibroblastic effects when applied after corneal alkali burns in rat corneas. Further studies should be conducted in this field.


Assuntos
Anti-Inflamatórios/uso terapêutico , Queimaduras Químicas/tratamento farmacológico , Neovascularização da Córnea/tratamento farmacológico , Queimaduras Oculares/tratamento farmacológico , Hypericum , Extratos Vegetais/uso terapêutico , Animais , Queimaduras Químicas/patologia , Neovascularização da Córnea/patologia , Queimaduras Oculares/induzido quimicamente , Queimaduras Oculares/patologia , Feminino , Fibroblastos/efeitos dos fármacos , Ratos Wistar
6.
Exp Eye Res ; 185: 107665, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31095932

RESUMO

Limbal stem cell deficiency (LSCD) is one of the serious cause of visual impairment and blindness with loss of corneal clarity and vascularization. Factors such as ocular burns (acids, lime, thermal), genetic disorders or infections results in the loss of limbal stem cells leading to LSCD. Reliable animal models of LSCD are useful for understanding the pathophysiology and developing novel therapeutic approaches. The purpose of the present study was to validate small and large animal models of LSCD by immunohistochemcal, clinical and histopathological comparison with human. The animal models of LSCD were created by topical administration of sodium hydroxide on the ocular surface of C57BL/6 mice (m, n = 12) and New Zealand white rabbits (r, n = 12) as per the standard existing protocol. Human corneal specimens (h, n = 12) were obtained from tissue bank who had chemical burn-induced LSCD. All samples were either paraffin embedded or frozen in cryogenic medium and the sections were processed for Hematoxylin-Eosin and Periodic Acid-Schiff staining to analyse the morphology and histopathological features of the corneal surface such as vascularization, inflammation, presence of goblet cells, epithelial hyperplasia and keratinization. Immunofluorescence was performed to distinguish between corneal (CK3+), conjunctival (CK19+) and epidermal (CK10+) epithelial phenotype. Histological analysis of corneal specimens from the three groups showed the presence of goblet cells (h:83%, m:50%, r:50%, p = 0.014), epithelial hypertrophy (h:92%, m:50%, r:66.6%, p = 0.04), epithelial hyperplasia (h:50%, m:17%, r:17%, p = 0.18), intra epithelial edema (h:42%, m:33%, r:100%, p = 0.02), stromal inflammation (h:100%, m:67%, r:67%, p = 0.01) and stromal vascularization (h:100%, m:50%, r:67%), in varying proportions. Immunostaining showed presence of total LSCD (CK19 + and/or CK10+, CK3-) in 92% of human and 50% of animal specimens. While partial LSCD (CK19 + and/or CK10+, CK3+) was seen in 8% of human and 50% of animal specimens. Our study shows the significant differences in the extent of vascularization, inflammation, epithelial thickness and goblet cell formation in mice and rabbit models of LSCD when compared to post-chemical burn LSCD in human corneas. In both mice and rabbit models complete LSCD developed in only 50% of cases and this important fact needs to be considered when working with animal models of LSCD.


Assuntos
Queimaduras Químicas/patologia , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Queimaduras Oculares/induzido quimicamente , Células Caliciformes/patologia , Ceratite/patologia , Limbo da Córnea/patologia , Animais , Queimaduras Químicas/metabolismo , Doenças da Córnea/metabolismo , Doenças da Córnea/patologia , Neovascularização da Córnea/metabolismo , Células Epiteliais/metabolismo , Epitélio Anterior , Queimaduras Oculares/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Células Caliciformes/metabolismo , Humanos , Imunofenotipagem , Inflamação/metabolismo , Inflamação/patologia , Queratina-19/metabolismo , Queratina-3/metabolismo , Ceratite/metabolismo , Limbo da Córnea/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucinas/metabolismo , Coelhos , Hidróxido de Sódio/toxicidade
7.
Cornea ; 38(6): 742-747, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30882541

RESUMO

PURPOSE: To detect early growth of blood and lymphatic vessels in the mouse cornea and iris after penetrating keratoplasty. METHODS: Penetrating keratoplasty was performed with C57BL/6 mice as donors and BALB/c mice as recipients. Graft transparency and neovascularization were examined by slit-lamp microscopy. Whole mounts of the cornea and iris were processed for detection of the outgrowth of blood and lymph vessels. RESULTS: On day 3 after surgery, all corneal grafts were slightly edematous, and blood vessels in the corneoscleral limbus dilated. LYVE-1 lymphatic vessels and CD31 blood vessels were distributed in the peripheral cornea. In the iris, the density of blood vessels increased, and LYVE-1 cells nearly vanished. On day 7, the grafts became opaque, and blood vessels grew into the recipient bed. A great quantity of lymph vessels invaded the cornea. LYVE-1 arborescent cells were found around the lymphatic vessels. In the iris, blood vessels became bulky and stiff, and arborescent LYVE-1 cells increased in number. On day 14, corneal neovascular regression and graft clarity were found. Lymphatic vessels regressed more slowly than blood vessels in the cornea. In the iris, blood vessels remained coarse. Increasing arborescent LYVE-1 cells were also noted in the ciliary body. CONCLUSIONS: Our findings suggest that the iris-ciliary body could amplify immune signals and in part promote initiation of immune rejection after keratoplasty by providing a pathway for macrophages, which might participate in corneal lymphangiogenesis.


Assuntos
Vasos Sanguíneos/patologia , Córnea/irrigação sanguínea , Rejeição de Enxerto/patologia , Iris/irrigação sanguínea , Ceratoplastia Penetrante/efeitos adversos , Vasos Linfáticos/patologia , Neovascularização Patológica/patologia , Animais , Corpo Ciliar/imunologia , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Rejeição de Enxerto/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Microvasos/patologia
8.
Nat Commun ; 10(1): 967, 2019 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-30814500

RESUMO

The stem cell pluripotency factor Oct4 serves a critical protective role during atherosclerotic plaque development by promoting smooth muscle cell (SMC) investment. Here, we show using Myh11-CreERT2 lineage-tracing with inducible SMC and pericyte (SMC-P) knockout of Oct4 that Oct4 regulates perivascular cell migration and recruitment during angiogenesis. Knockout of Oct4 in perivascular cells significantly impairs perivascular cell migration, increases perivascular cell death, delays endothelial cell migration, and promotes vascular leakage following corneal angiogenic stimulus. Knockout of Oct4 in perivascular cells also impairs perfusion recovery and decreases angiogenesis following hindlimb ischemia. Transcriptomic analyses demonstrate that expression of the migratory gene Slit3 is reduced following loss of Oct4 in cultured SMCs, and in Oct4-deficient perivascular cells in ischemic hindlimb muscle. Together, these results provide evidence that Oct4 plays an essential role within perivascular cells in injury- and hypoxia-induced angiogenesis.


Assuntos
Neovascularização Fisiológica , Fator 3 de Transcrição de Octâmero/deficiência , Células-Tronco Pluripotentes/metabolismo , Animais , Morte Celular , Linhagem da Célula , Movimento Celular , Células Cultivadas , Neovascularização da Córnea/metabolismo , Neovascularização da Córnea/patologia , Feminino , Membro Posterior , Isquemia/metabolismo , Isquemia/patologia , Masculino , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Miócitos de Músculo Liso/metabolismo , Neovascularização Patológica , Fator 3 de Transcrição de Octâmero/genética , Pericitos/metabolismo , Pericitos/patologia , Células-Tronco Pluripotentes/patologia
9.
Int J Mol Med ; 43(4): 1888-1899, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30816491

RESUMO

Hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF), two paracrine growth factors, modulate corneal epithelial cell metabolism, apoptosis and survival. Vascular endothelial growth factor (VEGF) serves as a proangiogenic factor in corneal neovascularization (CNV), which is a major cause of vision impairment and corneal blindness. The aim of the present study was to evaluate the ability of HGF and KGF to influence VEGF and its receptor, kinase insert domain receptor (Flk­1) in corneal injury and CNV in rats induced by ultraviolet radiation (UVR). An UVR­induced corneal injury rat model was successfully established to characterize the expression patterns of KGF, HGF, VEGF and Flk­1 in corneal tissues. Corneal epithelial cells were extracted and treated with small interfering RNAs (siRNAs) targeting KGF, HGF or both (si­KGF, si­HGF or si­HGF/KGF). The effects of HGF and KGF were examined through detection of the expression of KGF, HGF, VEGF and Flk­1, and the evaluation of cell proliferation, cell cycle and cell apoptosis. The expression levels of KGF, HGF, VEGF and Flk­1 in corneal tissues were increased in the rat model. In the cell experiments, the transfection of si­HGF/KGF resulted in reductions in VEGF, Flk­1, KGF and HGF. In addition, decreased cell proliferation and elevated cell apoptosis were found in the corneal epithelial cells from the rat model following KGF and HGF gene silencing. Taken together, these findings suggest that HGF and KGF gene silencing inhibits UVR­induced corneal epithelial proliferation and CNV and may function as novel targets for corneal wound healing.


Assuntos
Neovascularização da Córnea/genética , Neovascularização da Córnea/patologia , Fator 7 de Crescimento de Fibroblastos/genética , Inativação Gênica , Fator de Crescimento de Hepatócito/genética , Raios Ultravioleta , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Animais , Apoptose , Pontos de Checagem do Ciclo Celular , Proliferação de Células , Córnea/patologia , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Epitélio Anterior/metabolismo , Epitélio Anterior/patologia , Fator 7 de Crescimento de Fibroblastos/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Wistar
10.
BMC Ophthalmol ; 19(1): 67, 2019 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-30845992

RESUMO

BACKGROUND: To evaluate the ocular surface characteristics and the infestation of Demodex in Chinese paediatric and adult blepharokeratoconjunctivitis (BKC). METHODS: Fifty consecutive patients with BKC and 50 age- and sex-matched healthy subjects were enrolled. Lid margin characteristics and corneal disorders were evaluated under slit-lamp illumination. Four eyelashes were collected from each eye to examine Demodex infestation by light microscopy. RESULTS: Corneal neovascularization (P = 0.001) and scarring (P = 0.040) were significantly worse in children than in adults with BKC, whereas meibum quality was worse in adults (P = 0.008). Diagnosis delay was longer in children with BKC than in adults (2.2 vs 1.2 years, P = 0.022). Demodex infestation was more frequent in subjects with BKC than in healthy subjects (56% vs 26%, P = 0.002). The lid margin inflammation and meibomian gland dysfunction were worse in Demodex-positive subjects than in Demodex-negative subjects with BKC. CONCLUSIONS: Children with BKC had severer corneal disorders compared with adult BKC patients, which may be caused by a long-delayed diagnosis. Ocular demodicosis was more common in subjects with BKC. Ocular Demodex infestation was associated with worse lid margin inflammation and meibomian gland dysfunction.


Assuntos
Blefarite/parasitologia , Conjuntivite/parasitologia , Doenças da Córnea/patologia , Pestanas/parasitologia , Doenças Palpebrais/parasitologia , Infestações por Ácaros/complicações , Adolescente , Adulto , Estudos de Casos e Controles , Criança , Pré-Escolar , Doenças da Córnea/etiologia , Neovascularização da Córnea/patologia , Diagnóstico Tardio , Doenças Palpebrais/complicações , Pálpebras/patologia , Feminino , Humanos , Masculino , Glândulas Tarsais/patologia , Pessoa de Meia-Idade , Adulto Jovem
11.
Cutan Ocul Toxicol ; 38(4): 315-321, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30741024

RESUMO

Purpose: To compare the therapeutic effects of human derivatives in a mouse alkali burn model. Methods: The right eyes of mice were injured using NaOH. After alkali injury, one of the following agents was topically administered for 7 d: human amniotic membrane (hAM) suspension, human umbilical cord serum (hUCS), and human peripheral blood serum (hPBS), or saline. The epithelial defect areas on days 1, 2, and 3 degrees of opacity on days 2, 3, and 7, and corneal neovascularization (NV) areas on day 7 were evaluated. Histologic examination and mRNA expression levels of tumour necrosis factor (TNF)-α, interleukin (IL)-6, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, MMP-8, and MMP-9 were also evaluated on day 7. Results: The epithelial defect areas in the hUCS group were smaller than those in the control and hPBS groups on day 3 (p < .05, respectively). The epithelial defect areas in the hAM suspension group showed smaller than those in the control and hPBS groups on days 1 and 2 (p < .05, respectively). The degrees of opacity were lower in all treatment groups than that of the saline control group on day 7 (p < .05, respectively). Corneal NV areas were not different among groups on day 7 (p = 0.20). The expression levels of TNF-α, IL-6, MMP-8, and MMP-9 mRNA and the infiltration of the inflammatory cells in all treatment groups were lesser than those in the control group on day 7 (p< .05, respectively). Conclusions: All treatments reduced inflammatory reactions and corneal opacity development. Corneal reepithelialization was faster in the hUCS group.


Assuntos
Âmnio , Queimaduras Químicas/terapia , Neovascularização da Córnea/terapia , Opacidade da Córnea/terapia , Queimaduras Oculares/terapia , Soro , Hidróxido de Sódio/toxicidade , Animais , Queimaduras Químicas/patologia , Córnea/efeitos dos fármacos , Córnea/patologia , Neovascularização da Córnea/patologia , Opacidade da Córnea/patologia , Queimaduras Oculares/induzido quimicamente , Queimaduras Oculares/patologia , Humanos , Masculino , Camundongos Endogâmicos BALB C
12.
Burns ; 45(2): 398-412, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30600126

RESUMO

BACKGROUND: Since recent reports have shown that (-)-Epigallocatechin-3-gallate (EGCG) could be used for treating proliferative and inflammatory disorders, we explored its use for the management of corneal chemical burns. MATERIALS AND METHODS: Initially, EGCG was assayed on the rabbit corneal epithelial cell line RCE1(5T5) to establish the best testing conditions, and to avoid unwanted outcomes in the experimental animals. Then, we studied its effects on cell proliferation, cell cycle progression and cell differentiation. Afterwards, we instilled EGCG in experimental grade II corneal alkali burns in mice, three times a day up to 21days, and evaluated by slit lamp examination and histological sections of corneal epithelial, corneal endothelial and stromal edema, as well as the presence of inflammatory cells and neovascularization. RESULTS: EGCG reduced cell growth and led to a decline in the proportion of proliferative cells in a concentration dependent manner. At 10µM, EGCG promoted cell differentiation, an effect not related with apoptosis or cytotoxicity. When 10µM EGCG was instilled in corneal alkali burns in mice three times a day up to 21days, EGCG significantly reduced corneal opacity and neovascularization. The improved clinical appearance of the cornea was associated to a controlled epithelial growth; epithelial morphology was similar to that observed in normal epithelium and contrasted with the hyperproliferative, desquamating epithelium observed in control burn wounds. EGCG reduced corneal, stromal and endothelial edema, and wound inflammation. CONCLUSION: This work constitutes the first evidence for the use of EGCG in the acute phase of a corneal alkali burn, representing a possible novel alternative to improve patient outcomes as an add-on therapy.


Assuntos
Antioxidantes/farmacologia , Queimaduras Químicas/patologia , Catequina/análogos & derivados , Córnea/efeitos dos fármacos , Queimaduras Oculares/patologia , Cicatrização/efeitos dos fármacos , Álcalis/efeitos adversos , Animais , Catequina/farmacologia , Cáusticos/efeitos adversos , Ciclo Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Neovascularização da Córnea/patologia , Opacidade da Córnea/patologia , Epitélio Anterior/citologia , Epitélio Anterior/efeitos dos fármacos , Queimaduras Oculares/induzido quimicamente , Camundongos , Coelhos , Hidróxido de Sódio/efeitos adversos
13.
Int Ophthalmol ; 39(5): 1123-1135, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-29700650

RESUMO

PURPOSE: To evaluate the effect of metronidazole ophthalmic solutions on corneal neovascularization (CNV) in a rat model. METHODS: A chemical burn was created in the right central cornea of 40 rats. Animals were randomized and distributed into four study groups (n = 10 rats per group) designated Met_0.1%, Met_0.5%, sham, and untreated groups. Chemical-burned corneas in the Met_0.1% and Met_0.5% groups received ophthalmic solutions of 0.1 and 0.5% metronidazole, respectively. Corneas in the sham group received phosphate-buffered saline (metronidazole diluent). All treated eyes received ophthalmic solution at intervals of 6 h, for up to 30 days. Untreated corneas received no treatment. CNV was evaluated postinjury using corneal photographs at different evaluation time points. The main CNV outcome measures were: burn intensity, index of CNV, and percentage of vascularized corneal area. Five rats from each group were euthanized, on days 15 and 30; the samples were collected for histological analyses. Differences with P < 0.05 were considered significant. RESULTS: CNV was observed in the eyes from day 7 postinjury. However, the indices of CNV for the Met_0.1% and Met_0.5% groups were smaller than those for the sham and untreated groups (P < 0.05). Furthermore, corneas treated with 0.1 or 0.5% metronidazole had smaller vascularized areas compared to control corneas. On histological study, the presence of blood vessels confirmed clinical outcomes. CONCLUSIONS: Regular instillation of 0.1 or 0.5% metronidazole had a significant inhibitory effect for CNV on chemical burns induced in a rat model.


Assuntos
Córnea/patologia , Neovascularização da Córnea/tratamento farmacológico , Metronidazol/administração & dosagem , Animais , Anti-Infecciosos/administração & dosagem , Queimaduras Químicas/complicações , Córnea/efeitos dos fármacos , Neovascularização da Córnea/etiologia , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Queimaduras Oculares/complicações , Seguimentos , Masculino , Soluções Oftálmicas , Ratos , Ratos Wistar , Fatores de Tempo , Resultado do Tratamento
14.
Exp Eye Res ; 178: 177-185, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30321512

RESUMO

Toll-like receptors (TLRs) play an important role in inflammatory and immunological responses, which are intimately related to neovascularization. However, the precise mode of action of TLR3 in neovascularization still remains ambiguous. In this study, we sought to investigate the role of TLR3 in pathological corneal neovascularization (CNV) using a mouse model of alkali-induced CNV. CNV was attenuated in TLR3-deficient mice, and the absence of TLR3 led to decreased production of stromal cell-derived factor 1 (SDF-1), a well-characterized cytokine that regulates the recruitment of endothelial progenitor cells (EPCs) to the sites of neo-angiogenic niches in the injured tissues. Topical administration of polyinosinic-polycytidylic acid [poly (I:C)], a synthetic ligand for TLR3, to the injured cornea promoted CNV in wild type (WT) mice but not in TLR3-deficient mice. In addition, the effect of poly (I:C) on WT mice was abolished by addition of SDF-1 receptor antagonist AMD 3100. Furthermore, poly (I:C) treatment in vitro enhanced the migration of EPCs, whereas the enhanced migration was abolished by AMD 3100. These results indicate an essential role of TLR3 signalling in CNV that involves upregulating SDF-1 production and recruiting EPCs to the sites of injury for neovascularization. Thus, targeting the TLR3 signalling cascade may constitute a novel therapeutic approach for treating neovascularization-related diseases.


Assuntos
Quimiocina CXCL12/metabolismo , Neovascularização da Córnea/metabolismo , Células Progenitoras Endoteliais/citologia , Transdução de Sinais/fisiologia , Receptor 3 Toll-Like/metabolismo , Administração Oftálmica , Animais , Queimaduras Químicas/metabolismo , Movimento Celular/fisiologia , Córnea/efeitos dos fármacos , Neovascularização da Córnea/induzido quimicamente , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Queimaduras Oculares/induzido quimicamente , Técnica Indireta de Fluorescência para Anticorpo , Camundongos , Camundongos Endogâmicos C57BL , Poli I-C/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Hidróxido de Sódio/toxicidade
15.
Int Ophthalmol ; 39(7): 1553-1558, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30006905

RESUMO

PURPOSE: Fumagillin has been previously used to treat corneal microsporidial keratitis and also identified as an angiogenesis inhibitor. This study aimed to evaluate efficacy of fumagillin bicyclohexylamine on the rat model of corneal neovascularization induced by silver nitrate cauterization. METHODS: Twenty-four Albino Wistar rats (n = 24) were divided into three groups. Following silver nitrate-induced corneal injury, eyes in Group 1 received one drop of 5 mg/mL topical fumagillin bicyclohexylamine four times daily for 10 days. Group 2 received subconjunctival injection of 0.1 mL fumagillin bicyclohexylamine (2.5 mg/mL) on day 1 and day 5. Group 3 received artificial tears and lubricants four times daily for 10 days as control. On day 10, animals were sacrificed. Corneal specimens were obtained and prepared to assess vascular endothelial growth factor (VEGF-C) levels and corneal angiogenic microvessel density. RESULTS: There was no significant difference in VEGF-C levels between the groups (P = 0.994). Assessment of angiogenic microvessel density for peripheral corneal zone also did not reveal significant difference between the groups (P = 0.113). However, mean vascular density in Group 1 and Group 2 was significantly higher for both midperipheral and central corneal zones in comparison with Group 3 (P = 0.003, P = 0.015). CONCLUSIONS: Previously proved to be effective for treatment of microsporidial keratitis in humans, topical and subconjunctival concentration or dosing of fumagillin bicyclohexylamine failed to reduce corneal neovascularization induced by silver nitrate in this study. Further studies comparing different concentrations and dosing may detect inhibitory effects of fumagillin on corneal neovascularization without inducing toxicity.


Assuntos
Córnea/irrigação sanguínea , Neovascularização da Córnea/tratamento farmacológico , Cicloexanos/administração & dosagem , Ácidos Graxos Insaturados/administração & dosagem , Administração Tópica , Inibidores da Angiogênese/administração & dosagem , Animais , Túnica Conjuntiva , Córnea/efeitos dos fármacos , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Injeções , Soluções Oftálmicas , Distribuição Aleatória , Ratos , Ratos Wistar , Sesquiterpenos/administração & dosagem , Resultado do Tratamento
16.
Biosci Rep ; 39(1)2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30355648

RESUMO

Expression of hypoxia-inducible factor (HIF) 1α has been observed in corneal neovascularization (CNV). Vascular endothelial growth factor (VEGF), one of the most well-known angiogenic factors in CNV, is under the regulation of HIF-1. The present study aims to investigate the synergistic effects of VEGF and HIF-1α gene silencing on alkali burn-induced CNV in rabbits. The models of rabbits in corneal alkali burn were established. SiRNA recombinant adenovirus was used to explore the synergistic effects of VEGF and HIF-1α gene silencing on alkali burn-induced CNV. CNV area and ultrastructure of cornea were observed. The expression of VEGF and HIF-1α was detected. CNV was observed in rabbits following alkali burn. In addition, overexpressed VEGF and HIF-1α was also observed in rabbits following alkali burn. Then, silencing HIF-1α or silencing VEGF decreased area of CNV, inhibited neovascularization and improved pathological changes, while double-target interference for VEGF and HIF-1α decreased area of CNV inhibited neovascularization, and improved pathological changes to a greater extent. Our study provides evidences emphasizing the distinct notion that VEGF and HIF-1α play the contributory role in alkali burn-induced CNV as a result of double-target interference for VEGF and HIF-1α inhibiting CNV in rabbits following corneal alkali burn.


Assuntos
Queimaduras Químicas/complicações , Neovascularização da Córnea/etiologia , Queimaduras Oculares/complicações , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Neovascularização da Córnea/genética , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Células Epiteliais/patologia , Feminino , Inativação Gênica , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Coelhos , Fator A de Crescimento do Endotélio Vascular/metabolismo
17.
Mol Med Rep ; 18(5): 4388-4398, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30221697

RESUMO

The present study aimed to investigate the effects of diabetes mellitus (DM) on the generation of experimental corneal neovascularization (CrNV) and choroidal neovascularization (ChNV). Diabetes was induced in mice by intraperitoneal injection of streptozotocin (STZ). Experimental CrNV and ChNV were induced by alkali injury and laser photocoagulation, respectively. CrNV and ChNV were compared between the STZ­induced diabetic mice and control mice two weeks after injury. Relative expression of angiogenic factors was quantified by reverse transcription­quantitative polymerase chain reaction, and progenitor cell or macrophage accumulation in the early phase following injury was examined by flow cytometric analysis. Compared with the alkali­injured normal mice, the alkali­injured diabetic mice (STZ­induced) exhibited no significant difference in CrNV occurrence, whereas the laser­injured diabetic mice exhibited significantly reduced levels of ChNV compared with those of the laser­injured control animals. The laser­induced intrachoroidal mRNA expression levels of angiogenic factors, including vascular endothelial growth factor, hypoxia­induced factor­1α, chemokine (C­C motif) ligand 3, and stromal cell­derived factor­1α, were reduced in the laser­injured diabetic mice when compared with laser­injured control mice. Furthermore, the laser­induced intrachoroidal infiltration of c­Kit+ progenitor cells was impaired in the laser­injured diabetic mice compared with the laser­injured control mice. Overall, diabetes did not exert a significant effect on the generation of experimental CrNV. However, diabetes reduced laser­induced ChNV through downregulation of intrachoroidal progenitor cell infiltration and angiogenic factor expression.


Assuntos
Indutores da Angiogênese , Neovascularização de Coroide/genética , Neovascularização da Córnea/genética , Diabetes Mellitus Experimental/genética , Álcalis/toxicidade , Animais , Quimiocina CXCL12/genética , Neovascularização de Coroide/induzido quimicamente , Neovascularização de Coroide/complicações , Neovascularização de Coroide/patologia , Córnea/crescimento & desenvolvimento , Córnea/efeitos da radiação , Neovascularização da Córnea/complicações , Neovascularização da Córnea/etiologia , Neovascularização da Córnea/patologia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/patologia , Modelos Animais de Doenças , Citometria de Fluxo , Regulação da Expressão Gênica/genética , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Lasers/efeitos adversos , Camundongos , Fator A de Crescimento do Endotélio Vascular/genética
18.
Biomed Pharmacother ; 107: 1056-1063, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30257317

RESUMO

Antibody-based therapy is an effective strategy for treating ocular angiogenesis. However, short-acting efficacy and poor treatment compliance usually occurs in clinical practices. Thus, it is required to develop a drug delivery system to improve the bioavailability and decrease the toxicity of anti-angiogenic antibody. Bevacizumab is a recombinant humanized monoclonal antibody against vascular endothelial growth factor (VEGF). In this study, bevacizumab was encapsulated into poly (lactide-co-glycolide) (PLGA) nanoparticles. PLGA encapsulation could prolong the residency of bevacizumab in the vitreous and aqueous humor and produce long-lasting drug concentrations. Bevacizumab-encapsulated PLGA had no significant cytotoxicity and tissue toxicity effect in vitro and in vivo. In vitro studies showed that bevacizumab-encapsulated PLGA was more effective than bevacizumab in inhibiting VEGF-mediated endothelial cell proliferation, migration and tube formation. In vivo studies showed that bevacizumab-encapsulated PLGA enhanced the anti-angiogenic efficiency of bevacizumab for treating corneal neovascularization and retinal neovascularization. Thus, bevacizumab-encapsulated PLGA could increase the bioavailability and decrease the toxicity of bevacizumab during ocular angiogenesis therapy.


Assuntos
Inibidores da Angiogênese/administração & dosagem , Bevacizumab/administração & dosagem , Neovascularização da Córnea/tratamento farmacológico , Neovascularização Retiniana/tratamento farmacológico , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/toxicidade , Animais , Humor Aquoso/metabolismo , Bevacizumab/farmacologia , Bevacizumab/toxicidade , Disponibilidade Biológica , Neovascularização da Córnea/patologia , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nanopartículas , Tamanho da Partícula , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Neovascularização Retiniana/patologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Corpo Vítreo/metabolismo
20.
Mol Med Rep ; 18(3): 3203-3210, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30066863

RESUMO

The aim of the present study was to investigate the effect of microRNA 146a (miR146a) on promoting the repair of corneal alkali burn with bone marrow mesenchymal stem cells (MSCs). A total of 24 Sprague­Dawley female rats were divided into a normal group (Control), a normal MSC treatment group (Normal MSCs), an miR146a knockout MSC treatment group (miR146a­low MSCs) and an miR146a high­expression MSC treatment group (miR146a­high MSCs) according to the random number table. Quantitative polymerase chain reaction was used to evaluate the expression levels of miR146a. MTT assay was performed to measure the cell viability of mesenchymal stem cells (MSCs) and apoptosis was measured by flow cytometry. The expression levels of p65 nuclear factor (NF)­κB, proliferating cell nuclear antigen (PCNA) and Fas proteins were analyzed by western blotting. MSCs were tested for the secretion levels of vascular endothelial growth factor (VEGF), CD45, interferon (IFN)­Î³ and interleukin (IL)­10 by ELISA. The miR146a­high MSCs improved cell viability of MSCs and inhibited apoptosis of MSCs following alkali burn. miR146a­high MSCs decreased the expression levels of p65NF­κB and PCNA, and enhanced the expression level of Fas. Furthermore, miR146a­high MSCs improved the cornea opacity and enhanced the inhibition of neovascularization in the rats following alkali burn. miR146a­high MSCs inhibit the expression of VEGF, CD45, IFN­Î³, while enhanced the expression of IL­10. Therefore, miR146a promotes the repair of corneal alkali burn in rats treated with MSCs.


Assuntos
Queimaduras Químicas/terapia , Queimaduras Oculares/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , Regulação para Cima , Álcalis/efeitos adversos , Animais , Apoptose , Queimaduras Químicas/genética , Queimaduras Químicas/patologia , Sobrevivência Celular , Células Cultivadas , Córnea/patologia , Neovascularização da Córnea/genética , Neovascularização da Córnea/patologia , Neovascularização da Córnea/terapia , Modelos Animais de Doenças , Queimaduras Oculares/genética , Queimaduras Oculares/patologia , Feminino , Técnicas de Inativação de Genes , Células-Tronco Mesenquimais/citologia , Ratos , Ratos Sprague-Dawley
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