RESUMO
Diabetic retinopathy (DR) is a common microvascular complication that causes visual impairment or loss. Aquaporin 4 (AQP4) is a regulatory protein involved in water transport and metabolism. In previous studies, we found that AQP4 is related to hypoxia injury in Muller cells. Transient receptor potential cation channel subfamily V member 4 (TRPV4) is a non-selective cation channel protein involved in the regulation of a variety of ophthalmic diseases. However, the effects of AQP4 and TRPV4 on ferroptosis and oxidative stress in high glucose (HG)-treated Muller cells are unclear. In this study, we investigated the functions of AQP4 and TRPV4 in DR. HG was used to treat mouse Muller cells. Reverse transcription quantitative polymerase chain reaction was used to measure AQP4 mRNA expression. Western blotting was used to detect the protein levels of AQP4, PTGS2, GPX4, and TRPV4. Cell count kit-8, flow cytometry, 5,5',6,6'-tetrachloro-1,1,3,3'-tetraethylbenzimidazolyl carbocyanine iodide staining, and glutathione (GSH), superoxide dismutase (SOD), and malondialdehyde (MDA) kits were used to evaluate the function of the Muller cells. Streptozotocin was used to induce DR in rats. Haematoxylin and eosin staining was performed to stain the retina of rats. GSH, SOD, and MDA detection kits, immunofluorescence, and flow cytometry assays were performed to study the function of AQP4 and TRPV4 in DR rats. Results found that AQP4 and TRPV4 were overexpressed in HG-induced Muller cells and streptozotocin-induced DR rats. AQP4 inhibition promoted proliferation and cell cycle progression, repressed cell apoptosis, ferroptosis, and oxidative stress, and alleviated retinal injury in DR rats. Mechanistically, AQP4 positively regulated TRPV4 expression. Overexpression of TRPV4 enhanced ferroptosis and oxidative stress in HG-treated Muller cells, and inhibition of TRPV4 had a protective effect on DR-induced retinal injury in rats. In conclusion, inhibition of AQP4 inhibits the ferroptosis and oxidative stress in Muller cells by downregulating TRPV4, which may be a potential target for DR therapy.
Assuntos
Aquaporina 4 , Retinopatia Diabética , Células Ependimogliais , Ferroptose , Estresse Oxidativo , Canais de Cátion TRPV , Animais , Masculino , Camundongos , Ratos , Aquaporina 4/metabolismo , Aquaporina 4/genética , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Retinopatia Diabética/metabolismo , Retinopatia Diabética/patologia , Retinopatia Diabética/genética , Células Ependimogliais/metabolismo , Células Ependimogliais/patologia , Glucose/metabolismo , Glucose/farmacologia , Camundongos Endogâmicos C57BL , Ratos Sprague-Dawley , Canais de Cátion TRPV/metabolismo , Canais de Cátion TRPV/genéticaRESUMO
Diabetic retinopathy (DR) is one of the most serious microvascular complications of diabetes and an important cause of blindness in adults. In previous study, we found that miR-320a alleviated the damage of muller cells in DR. In this study, we mainly explored the mechanism of lncRNA MALAT1 on retinal angiogenesis in DR by regulating miR-320a/HIF-1α. The expression of MALAT1 and miR-320a was detected by RT-qPCR, and the expression of HIF-1α was detected by Western blot. The superoxide anion level, invasion, angiogenesis, and vascular permeability of mouse retinal microvascular endothelial cells (MRMECs) co-cultured with muller cells were evaluated by dihydroethidium, transwell, angiogenesis and immunofluorescence assay. In order to analyze the targeting relationship between miR-320a and MALAT1 or HIF-1α, we performed dual luciferase reporter gene, fluorescence in situ hybridization (FISH), RNA immunoprecipitation (RIP) and RNA pulldown experiments. The results should that MALAT1 and HIF-1α were highly expressed and miR-320a was low expressed in high glucose (HG)-induced muller cells, and MALAT1 could competitively bind with HIF-1α. Knocking down miR-320a inhibited MRMECs invasion angiogenesis, and vascular permeability by targeting miR-320a. Overexpression of miR-320a down regulated HIF-1α and inhibited the invasion, angiogenesis, and vascular permeability of MRMECs. In conclusion, MALAT1 inhibits HIF-1α expression and MRMECs angiogenesis in DR through spongy miR-320a.
Assuntos
Diabetes Mellitus , Retinopatia Diabética , MicroRNAs , RNA Longo não Codificante , Animais , Diabetes Mellitus/metabolismo , Retinopatia Diabética/genética , Retinopatia Diabética/metabolismo , Células Endoteliais/metabolismo , Hibridização in Situ Fluorescente , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Neovascularização Patológica/metabolismo , RNA Longo não Codificante/genéticaRESUMO
INTRODUCTION: The main aim of this article was to study the retinal peripapillary and macular vascular structures in eyes with primary angle-closure suspects (PACS) using optical coherence tomography angiography (OCTA). METHODS: In this cross-sectional study, control and PACS subjects were recruited from a community screening. Only one eye per subject was used for analysis. All participants underwent a questionnaire survey, physical and ophthalmic examinations, ocular biometry measurements, and OCTA. We compared basic demographics and vessel structure parameters between control and PACS eyes. Univariate and multivariate linear regression analyses were performed to investigate factors associated with vascular parameters in both groups. RESULTS: Data from 254 subjects including 155 PACS and 99 controls were analyzed. In the peripapillary region, PACS eyes showed similar retina nerve fiber layer (RNFL) and vessel densities (VDs) including and excluding large vessels compared to control eyes. Compared to control eyes, all macular OCTA parameters showed significant differences in PACS eyes, including decreased superficial VD (p = 0.006) and deep VD (p = 0.004), larger fovea avascular zone (FAZ) area (p = 0.006), and longer FAZ perimeter (p = 0.004). Gender (p = 0.039), age (p < 0.001), and Garway-Heath superior hemisphere RNFL (p < 0.001) were risk factors influencing optic disc VD excluding large vessels. Axial length was the major factor affecting macula superficial and deep VDs (p = 0.004 and 0.001 respectively), while PACS was an independent factor associated with larger FAZ perimeter (p = 0.046). CONCLUSION: While PACS and control eyes have comparable RNFL and vascular structure around the optic nerve head, macular vascular structures are significantly different.
Assuntos
Macula Lutea , Disco Óptico , Humanos , Estudos Transversais , Macula Lutea/irrigação sanguínea , Macula Lutea/diagnóstico por imagem , Disco Óptico/irrigação sanguínea , Disco Óptico/diagnóstico por imagem , Vasos Retinianos/diagnóstico por imagem , Tomografia de Coerência Óptica/métodos , Angiografia/métodosRESUMO
INTRODUCTION: This study aimed to investigate the ganglion cell complex (GCC) parameters in primary angle closure suspects (PACS) and identify the related factors. METHODS: A total of 731 subjects, including 289 subjects with PACS and 442 subjects without PACS, underwent RTVue XR OCT. GCC parameters were compared between the two groups. The linear mixed-effects model was performed to evaluate the relationships between the GCC parameters and related factors. RESULTS: Significant differences were found in gender, age, spherical refractive error, height, waist, anterior chamber depth, lens thickness, axial length, superior GCC thickness, ganglion cell complex focal loss volume, ganglion cell complex global loss volume, and ganglion cell complex root mean square between PACS and normal controls. The linear mixed-effects model showed that age (p = 0.008) and blood glucose (p = 0.001) were negatively correlated with average GCC thickness in PACS subjects, and PACS (p = 0.036) and age (p < 0.001) were the key influencing factors for average GCC thickness. CONCLUSION: GCC parameters in PACS subjects are different from those in normal controls. Careful explanation should be considered when evaluating changes of GCC parameters in patients with PACD.
Assuntos
Células Ganglionares da Retina , Glaucoma de Ângulo Fechado , Gonioscopia , Humanos , Pressão Intraocular , Tomografia de Coerência ÓpticaRESUMO
Background: Ultraviolet (UV) exposure is the most essential etiological factor in sebaceous gland carcinoma (SGC). The abnormal expression of microRNAs (miRNAs) is also involved in SGC. However, the function of miRNAs in UV-induced SGC is still unclear. Methods: In this study, the expression levels of miR-651-5p and zinc finger E-box binding homeobox 2 (ZEB2) in SGC tissues and cells were measured by real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting. Then, the effects of miR-651-5p on the apoptosis, migration, invasion, and epithelial-mesenchymal transition (EMT) of UV-induced SGC cells were determined. The interactions between miR-651-5p and ZEB2 were verified by a dual-luciferase reporter assay. An in vivo tumor growth assay was performed to assess tumorigenicity. Results: The results showed that there was abnormal expression of miR-651-5p and ZEB2 in SGC tissues and cells compared with the control tissues and cells. Overexpression of miR-651-5p and knockdown of ZEB2 inhibited the malignant biological behaviors of SGC cells. Moreover, ZEB2 is one of the target genes of miR-651-5p, and the expression of ZEB2 was negatively regulated by miR-651-5p in SGC cells. Further studies showed that overexpression of miR-651-5p promoted cell apoptosis and inhibited the cell invasion and migration ability and EMT of UV-induced SGC cells by downregulating the expression of ZEB2 in vitro and in vivo. Conclusions: This study revealed that overexpression of miR-651-5p inhibited UV-induced SGC growth and metastasis by suppressing ZEB2, which may be a potential target for SGC prevention and therapy.
RESUMO
AIM: To compare visual field defects using the Swedish Interactive Thresholding Algorithm (SITA) Fast strategy with SITA Faster strategy, a newly developed time-saving threshold visual field strategy. METHODS: Ninety-three participants (60 glaucoma patients and 33 normal controls) were enrolled. One eye from each participant was selected randomly for the study. SITA Fast and SITA Faster were performed using the 24-2 default mode for each test. The differences of visual field defects between the two strategies were compared using the test duration, false-positive response errors, mean deviation (MD), visual field index (VFI) and the numbers of depressed test points at the significant levels of P<5%, <2%, <1%, and <0.5% in probability plots. The correlation between strategies was analyzed. The agreement between strategies was acquired by Bland-Altman analysis. RESULTS: Mean test durations were 246.0±60.9s for SITA Fast, and 156.3±46.3s for SITA Faster (P<0.001). The test duration of SITA Faster was 36.5% shorter than SITA Fast. The MD, VFI and numbers of depressed points at P<5%, <2%, <1%, and <0.5% in probability plots showed no statistically significant difference between two strategies (P>0.05). Correlation analysis showed a high correlation for MD (r=0.986, P<0.001) and VFI (r=0.986, P<0.001) between the two strategies. Bland-Altman analysis showed great agreement between the two strategies. CONCLUSION: SITA Faster, which saves considerable test time, has a great test quality comparing to SITA Fast, but may be not directly interchangeable.