Objective To observe and analyze the incidence of macular edema and its related factors after cataract surgery in diabetics with and without diabetic retinopathy. Methods A retrospective study. The data of 90 diabetics including 45 cases with diabetic retinopathy (DR group) and 45 cases without DR (diabetics group) and an equal number of non-diabetic matched controls (control group) who underwent phacoemulcification and intraocular lens implantation were collected. Patients with macular edema before the surgery were excluded. Main outcome measurements included best-corrected visual acuity (BCVA) and central subfield mean thickness (CSMT). Optical coherence tomography (OCT) was used to measure the distance from the inner limiting membrane to the pigment epithelium of the central macular with diameter of 1 mm, which was used as the CSMT. There were no significant differences in BCVA and CSMT among three groups preoperatively (F=1.300, 1.329; P=0.280, 0.273). The BCVA and CSMT before and after the surgery in all three groups were compared. macular edema was defined as an increase of CSMT on OCT ＞30% from preoperative baseline. The incidence of macular edema of three groups after the surgery were compared and analyzed. The correlation between postoperative BCVA and CSMT, and the correlation between diabetes mellitus, DR and macular edema after surgery were analyzed by Logistic regression analysis. Results After the surgery, compared with control and diabetics group, the BCVA in DR group decreased and the CSMT increased significantly and the differences were statistically significant (P＜0.05). However, between control and diabetics group, the differences in BCVA and CSMT after the surgery were not statistically significant (P＞0.05). The incidences of macular edema in DR group (15.6% and 13.3%) 1 month and 3 months postoperatively were significantly more than that in control group (2.2% and 2.2%) and non-DR diabetics group (4.4% and 2.2%), and the differences were statistically significant (χ2=6.696, 6.644; P=0.035, 0.036). Logistic regression analysis showed that the postoperative BCVA was correlated with CSMT (r=0.444, P=0.000), diabetics was not correlated with postoperative macular edema (r=7.231, P=0.999) and DR was correlated with macular edema after surgery (r=0.378, P=0.008). The diabetic retinopathy might correlated to macular edema after surgery. Conclusions The incidence of macular edema after cataract surgery in patients with DR was significantly higher than that in patients without DR. There is no correlation between diabetics and postoperative macular edema, and DR is correlated with macular edema after surgery.
Objective To observe the effect of celastrol on the secretion of interleukin (IL)-17 in peripheral blood mononuclear cells in patients with sympathetic ophthalmia (SO), and its possible mechanisms. Methods Venous blood samples were extracted from 10 cases of sympathetic ophthalmia patients and 10 health objectives. The peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation and then were divided into 4 groups. Group A (control group): PBMCs of health objectives; Group B: PBMCs of SO patients; Group C: PBMCs of SO patients with 0.5 μmol/L celastrol in the medium; Group D: PBMCs of SO patients with 1 μmol/L celastrol in the medium. After culturing the cells for 3 days, the supernatant of 4 groups was collected, and the levels of IL-23 and IL-17 were detected by enzyme-linked immuno sorbent assay (ELISA). Then, the 50 ng/ml rIL-23 was added into the medium of group A which was the group A1; the 50ng/ml rIL-23 and 1 μmol/L Cela were added into to the medium of group A which was the group A2. For the medium of group B, the 50 ng/ml rIL-23 was added into the medium which was the group B1; the 50 ng/ml rIL-23 and 1 μmol/L celastrol were added into to the medium of group B which was the group B2. After culturing for 3 days, the supernatant of cells of these 4 groups was collected, and the levels of IL-17 were detected by ELISA. Results In group A, the levels of IL-23 and IL-17 were (228.43±17.27) pg/ml and (220.55±31.15) pg/ml respectively. In group B, the levels of IL-23 and IL-17 were (513.85±36.46) pg/ml and (866.77±72.92) pg/ml respectively. In group C, the levels of IL-23 and IL-17 were (381.07±20.93) pg/ml and (517.43±54.87) pg/ml respectively. In group D, the levels of IL-23 and IL-17 were (237.14±17.97) pg/ml and (242.89±34.09) pg/ml respectively. Between group A and D, there was no statistically significant difference in IL-23 or IL-17 level (P＞0.05); but when comparing other groups, the differences were statistically significant (P＜0.05). The levels of IL-17 in group A1 and group A2 were (428.43±24.53) pg/ml and (229.15±23.28) pg/ml and the difference was statistically significant (P＜0.05). The levels of IL-17 in group B1 and group B2 were (1373.39±89.51) pg/ml and (571.01±94.88) pg/ml and the difference was statistically significant (P＜0.05). Conclusion Celastrol can inhibit the secretion of IL-17 by PBMCs in SO patients via inhibiting the secretion of IL-23.
ObjectiveTo observe the expression of miR-142-5p and forkhead transcription protein O subgroup 3 (FOXO3) in CD4+ T cells of experimental autoimmune uveitis (EAU) model rats, and preliminarily explore the targeting relationship between the two and the effect on EAU impact.MethodsTen Lewis rats were randomly divided into model group and control group. Rats in the model group wree induced an EAU animal model by adoptive immunization. Twenty days after immunization, CD4+ T cells were extracted from the eyeballs and draining lymph nodes of rats in the control group and model group, and divided into control group, model group, mimic-negative control (NC) group, miR-142-5p-mimic group, and small interference (si)-NC group, si-FOXO3 group for in vitro experiments. The miR-142-5p-mimic group and si-FOXO3 group were transfected with miR-142-5p-mimic and si-FOXO3, respectively. Twenty-five Lewis rats were randomly divided into model group, mimic-NC transfected group, miR-142-5p-mimic transfected group, si-NC transfected group, and si-FOXO3 transfected group. The above-mentioned in vitro experimental groups were injected with cells respectively. Slit lamp microscopy and EAU score were performed on 4, 8, 12, 16, 20 days after immunization; on 20 days after immunization, hematoxylin-eosin staining was performed for histopathological grading. Real-time fluorescence quantitative polymerase chain reaction was used to detect the relative expression of miR-142-5p and FOXO3 mRNA in CD4+ T cells and eye tissues of rats in each group, and helper T cell 17 (Th17) marker interleukin (IL)-17, IL-22, retinoic acid-related orphan receptor gamma (ROR gamma) relative expression level in the supernatant. Bioinformatics website and dual luciferase was used to predict the targeting relationship between miR-142-5p and FOXO3. One-way analysis of variance or t test was used for comparison between groups.ResultsAll rats in the model group showed symptoms of EAU to varying degrees, and the symptoms became worse with time. Compared with the control group, the relative expression of miR-142-5p mRNA in CD4+ T cells of the model group increased, and the relative expression of FOXO3 mRNA decreased. The differences were statistically significant (t=7.374, 10.423; P=0.002, 0.001). Compared with the mimic-NC group, the relative expression of miR-142-5p mRNA in the CD4+ T cells of the miR-142-5p-mimic group increased, and the difference was statistically significant (t=6.540, P=0.003). Compared with the model group, mimic-NC group, and si-NC group, the relative expression of IL-17, IL-22, and RORγ mRNA in CD4+ T cells in the miR-142-5p-mimic group and si-FOCO3 group increased significantly. The difference was statistically significant (F=26.110, 6.292, 5.269, 55.660, 10.490, 11.430; P＜0.05). Compared with the mimic-NC transfected group, the relative expression of miR-142-5p mRNA in the ocular tissues of the miR-142-5p-mimic transfected rats increased significantly, and the difference was statistically significant (t=6.690, P＜0.05). Compared with the transfected si-NC group, the relative expression of FOXO3 mRNA in the eye tissue of the transfected si-FOXO3 group was significantly decreased, and the difference was statistically significant (t=17.751, P＜0.05). Rats in the mimic-NC transfected group, miR-142-5p-mimic transfected group, si-NC transfected group, and si-FOXO3 transfected group prolonged with time after immunization, and the EAU scores showed an upward trend. The EAU score and histopathological grade of rats in the miR-142-5p-mimic transfected group were higher than those in the mimic-NC transfected group, and the difference was statistically significant (t=5.633, 6.286; P＜0.05). The EAU score and histopathological grade of the rats in the transfected si-FOXO3 group were higher than those in the transfected si-NC group, and the difference was statistically significant (t=6.852, 6.635; P＜0.05). FOXO3 has a targeting relationship with miR-142-5p.ConclusionsIn EAU rat CD4+ T cells, the expression of miR-142-5p is up-regulated, while the expression of FOXO3 is down-regulated. miR-142-5p targets the expression of FOXO3 to promote the development of Th17 cell-related inflammatory factors.
ObjectiveTo observe the expressions of miR-183 and retinal dehydrogenase 11 (RDH11) in exosomes derived from bone marrow mesenchymal stem cells (BMSC), and to preliminarily explore their targeting relationship and their effects on retinal pigment epithelial (RPE) cells. MethodsBMSC from C57BL/6 (C57) mice were isolated and cultured, and BMSC-derived exosomes were identified. BMSC were divided into blank group, simulation blank control group (mimic-NC group), miR-183 simulation group (miR-183-mimic group). C57 mice and retinal degeneration 10 (rd10) mouse RPE cells were cultured with reference to literature methods. RPE cells from rd10 mice were transfected with BMSC exosomes and co-cultured and divided into control group, exosome group, mimic-NC-exosome group (mimic-NC-exo group), miR-183-mimic-exosome group (miR-183-mimic-exo group). The relative expression levels of miR-183, RDH11 mRNA and protein in C57 mice, rd10 mice and RPE cells in each group were detected by real-time quantitative polymerase chain reaction and western blotting. The targeting relationship between miR-183 and RDH11 was analyzed by bioinformatics website and dual luciferase reporter. Cell counting kit 8 was used to detect the effect of miR-183 on BMSC exosomes on RPE cell proliferation; in situ labeling end labeling method was used to detect RPE cells apoptosis. One-way ANOVA was used to compare multiple groups. ResultsCompared with C57 mouse RPE cells, the relative expression of miR-183 in rd10 mouse RPE cells was down-regulated, and the relative expression of RDH11 mRNA was up-regulated, and the differences were statistically significant (t=5.230, 8.548; P=0.006, 0.001). Compared with the blank group and the mimic-NC group, the relative expression of miR-183 mRNA in the exosomes of the miR-183-mimics group was significantly increased (F=60.130, P＜0.05). After 24 h of co-culture, exosomes entered RPE cells. Compared with the mimic-NC-exo group, the relative expression of miR-183 mRNA in RPE cells in the miR-183-mimic-exo group was significantly increased, the proliferation ability was enhanced (t=7.311, P=0.002), and the number of apoptotic cells was decreased (F=10.949, P=0.012), and the differences were statistically significant (t=4.571, P=0.002). Bioinformatics website and dual-luciferase report confirmed that miR-183 has a targeting relationship with RDH11. Compared with the mimic-NC group, the relative expression of RDH11 mRNA and protein in the exosomes of the miR-183-mimic group was decreased, and the difference was statistically significant (t=5.361, 6.591; P=0.006, 0.003). After co-culture, compared with the control group, there was no significant difference in the relative expression of RDH11 mRNA and protein in RPE cells in the exosome group (t=0.169, 1.134; P=0.874, 0.320); The relative expressions of RDH11 mRNA and protein in RPE cells in -183-mimic-exo group were decreased, and the difference was statistically significant (t=5.554, 5.546; P=0.005, 0.005). ConclusionUp-regulation of BMSC-derived exosomal miR-183 promote the proliferation of RPE cells in vitro by targeting the expression of RDH11 and reduce the number of apoptosis.