| OBJECTIVE:The current relationship between dysfunction and morphologic changes in the early stage of diabetic retinopathy is not completely clear.In this study,Angiography optical coherence tomography techniques were used to examine the circulation and structure of the retina in the macula,and microperimetry were used to detect the light sensitivity and fixation status of the macula.Through the combination of the two techniques,we explored the relationship between functional dysfunction and morphological changes in early diabetic retinopathy.METHODS:1 n Detection of OCT A and microperimetry on diabetic patients without diabetic retinopathy.Thirty-two eyes of 32 patients without diabetic retinopathy were collected,and 18 of 18 healthy volunteers matched with age were used as controls.All patients underwent basic examination of the eye.Angio-OCT was used to detect the retina thickness and the depth of the superficial and deep blood flow density in the 10°area of the macular center.Microperimetry was used to detect mean sensitivity in the 10°area of the macular center and 2°/40fixation rate.Quantitative data normal test,the independent sample t test was used to compare the two groups of patients with macular thickness,blood flow density,average sensitivity of the retina;macula morphology and functional relationship using Spearman correlation analysis,P values less than 0.05 were interpreted as statistically significant.2n Detection of OCT A and microperimetry on non-proliferative diabetic retinopathy patients.Twelve patients(24 eyes)with NDR and 12 patients with non-proliferative diabetic retinopathy(NPDR)were enrolled in this study.Twenty-four eyes of 13 healthy volunteers matched with age were used as controls.Angio-OCT was used to detect FAZ,the retina thickness and the depth of the superficial and deep blood flow density in the 10°area of the macular center.Microperimetry was used to detect mean sensitivity in the 10°area of the macular center.2°/4°fixation rate and BCEA.Measurement data were skewed distribution.Kruskal-Wallis test was used to compare the thickness,blood flow density,FAZ area,average retinal sensitivity,fixation and other indicators,Mann-Whitney u test for comparison between groups.Macular morphology and functional relationship using Spearman correlation analysis.RESULTS:1.1 The retinal thickness of the macular area in the NDR group was thicker than that in the control group,but there was no statistical difference.1.2 There was no significant difference in the superficial retinal blood flow density between the NDR group and the control group.1.3 Deep retinal blood flow density in the NDR group decreased compared with the control group,but there was no statistical difference.1.4 The mean retinal sensitivity in NDR group was significantly lower than that in control group,except for the inferior quadrant,with statistical difference.1.5 Correlation analysis showed that in the inferior quadrant,there was a significant correlation between retinal thickness and average retinal sensitivity under NDR group.There was a significant correlation between superficial retinal blood flow density and average retinal sensitivity in the superior and nasal quadrant in NDR group.2.1 The central retinal thickness of NPDR group was significantly thicker than that of control group.There was no significant differences im retinal thickness when we compared the NPDR group with the NDR group and the NDR group with the normal control group.2.2 Vessel flow density(superficial,deep)in the NPDR group was significantly lower than that in the control group.The deep blood flow density in the NPDR group was significantly lower than that in the NDR group.There was no statistical difference between the other two groups.2.3 The NPDR group had significantly larger FAZ than the control and NDR groups.There was no statistical difference between the NDR group and the control group in FAZ;2.4 The retinal mean sensitivity of the NPDR group was significantly lower than that in the normal control group and the NDR group.There was no statistical difference between the NDR group and the control group in the retinal mean sensitivity;2.5 2°,40fixation stability and BCE A in the NPDR group was significantly lower than that in the control group.There was no statistical difference between the NDR group and the control group in 2°,4°fixation stability and BCEA.CONCLUSION:(l)Changes in visual function preceded morphological changes in the early stage of diabetic retinopathy.(2)The mean retinal sensitivity is a very sensitive indicator to assess visual function changes in the early stage of diabetic retinopathy.(3)The average retinal sensitivity of retina in diabetic patients with no diabetic retinopathy correlates with retinal thickness and blood flow density,presenting a regional difference.(4)OCTA combined with microperimetry examination is an effective method to evaluate the morphological and functional changes of early stage of diabetic retinopathy.OBJECTIVE:Uric acid is an independent risk factor of diabetic retinopathy.Its pathogenesis in diabetic retinopathy is not yet clear.This experiment aims to explore the role of hyperuricemia in the development of diabetic retinopathy and the relationship between hyperuricemia and Notch signaling pathway by simulating the hyperuricemia and diabetes in vivo and in vitro in an attempt to elucidate the role of hyperuricemia through the regulation of Notch signaling Pathway to promote the development of diabetic retinopathy.Methods:1.In vitro experiments:1.1 Firstly,the models of hyperuricemia combined with high glucose,high uric acid and high glucose were constructed and the human retinal microvascular endothelial cells were incubated with high glucose/sodium mono urate crystal/high glucose combined with sodium mono urate respectively.1.2 The apoptosis rate of HRMECs in different treated cells was detected by Tunel staining.1.3 The mRNA levels of ICAM-1,IL-6,MCP-1,TNF-a,receptors Notch 1,ligands Dll 1,Dll 4,Jagged 1,Jagged 2 were detected by RT-PCR.1.4 Western-Blot was conducted to detect the expression of ICAM-1,IL-6,MCP-1,TNF-α,receptors Notch 1,ligands Dll 1,Dll 4,Jagged 1,Jagged 2.1.5 Notchl siRNA was used to interfere Notch signaling pathway,the expression of ICAM-1,IL-6,MCP-1 and TNF-α was detected by RT-PCR and Western blot respectively.2.In vivo experiments:2.1 Constructing high glucose,high uric acid and high glucose combined hyperuricemia model in vivo.Healthy male SD rats were injected intraperitoneally streptozotocin in the preparation of diabetic model,adenine and oteracil potassium combined gavage to prepare hyperuricemia model.Using automatic biochemical analyzer to detect blood glucose,uric acid,urea nitrogen,creatinine levels,to verify the success of modeling.2.2 RT-PCR and Western blot were used to detect the expression of inflammatory cytokines and the activity of Notch signaling pathway in rat retinas after different treatments.2.3 The mRNA levels of ICAM-1.IL-6.MCP-1,TNF-α,receptors Notch 1,ligands Dll 1,Dll 4,Jagged 1,Jagged 2 were detected by RT-PCR in rat retinas after different treatments.2.4 Western-Blot was conducted to detect the expression of ICAM-1,IL-6,MCP-1,TNF-α,receptors Notch 1,ligands Dll 1,Dll 4,Jagged 1,Jagged 2 in rat retinas after different treatments.3.Statistical methods:Results were expressed as mean±SD.Statistical analysis software SPSS 17.0 statistical software package and Graphpad Prism software system were used to process data.One-way ANOVA and Dunnett’s test were used to perform multiple comparisons between groups of variables.?P<0.05 represents a statistical difference.Results:(1)In vitro experiments:In high glucose,high uric acid and high glucose combined with high uric acid in vitro model,compared with the normal group,the apoptosis of HRMECs cells increased significantly,Among them,in the combination of high glucose and hyperuricemia group,the apoptosis of HRMECs was the most significant.Further detection of inflammatory cytokines in cells found that in high glucose,high uric acid and high glucose combined with hyperuricemia group,the expression of inflammatory cytokines were increased.Among them,the most obvious increase in inflammatory cytokines was in the combination of high glucose and hyperuricemia group.In the high glucose,high uric acid and high glucose combined with uric acid group,Notch signaling pathway activity were increased,Notch signaling pathway had the highest activity in the high glucose combined with uric acid group.After Notch 1 siRNA transfection in high glucose and high glucose combined with uric acid group,the activity of Notch signaling pathway was successfully down-regulated.We found that the apoptosis of HRMECs was significantly decreased in cells transfected with Notch 1 siRNA compared to the blank vector group,and the expression of inflammatory cytokines in cells was also significantly decreased.(2)In vivo experiments:Animal model of high uric acid,high uric acid and high glucose combined with hyperuricemia was successfully constructed.Compared with the control group,the expression of inflammatory cytokines in the retina of high glucose,high uric acid and high glucose combined with hyperuricemia rats increased,and the increase of inflammatory cytokines in hyperuricemic diabetic retinopathy rats was the most obvious.In addition,compared with the control group,the activity of Notch signaling pathway in the rat retina of high glucose,high uric acid and high glucose combined with hyperuricemia group was significantly increased.Moreover,the increase of Notch signaling pathway in hyperuricemic diabetic retinopathy rats was the most significant.3.Statistical methods:Results were expressed as mean±SD Statistical analysis software SPSS 17.0 statistical software package and Graphpad Prism software system were used to process data.One-way ANOVA and Dunnett’s test were used to perform multiple comparisons between groups of variables.,P<0.05 represents a statistical difference.Conclusion:(1)Both hyperglycemia and hyperuricemia can promote the inflammation of the retina.In addition,on the basis of high sugar,high uric acid can further promote the inflammation of the retina.(2)High glucose and high uric acid can increase the activity of Notch signaling pathway,while on the basis of high glucose,high uric acid can further increase Notch signaling pathway activity.(3)Hyperuricemia promotes the development of diabetic retinopathy by increasing the activity of Notch signaling pathway.(4)lowering uric acid is a potential treatment of diabetic retinopathy.(5)The activity of Notch signaling pathway can inhibit the inflammatory reaction of the retina.Notch signaling pathway is a potential therapeutic target for diabetic retinopathy. |
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