Potential Factors of Corneal Endothelial Cells for Progression in Children with Uveitis.

OBJECTIVE: To observe the morphological changes and abnormal structure of corneal endothelial cells in children with uveitis, to analyze the related factors affecting the morphological changes of corneal endothelial cells, and to explore the clinical application of a corneal endothelial microscope in children with uveitis. METHODS: The corneal endothelial cells of 70 patients with uveitis were photographed with the Topcon SP-3000 noncontact corneal endothelial microscope, and the corneal endothelial cell density (CD), average cell area (AVE), coefficient of variation of the cell area (CV), and percentage of hexagonal cells (PHC) were measured with the IMAGEnet system. Twenty-eight patients (56 eyes) with monocular uveitis were selected, with the affected eyes (28 eyes) as the experimental group and the contralateral healthy eyes (28 eyes) as the control group. The corneal endothelial cell parameters between the two groups were statistically analyzed. The parameters of corneal endothelial cells in 70 children with uveitis were compared, and the effects of the course of the disease, inflammatory cells in the anterior chamber, and posterior corneal deposition (KP) on the parameters of corneal endothelial cells were analyzed. RESULTS: There are four abnormal forms of the corneal endothelium in children with uveitis: enlarged cell area gap, irregular cell shape, blurred intercellular space, and cell loss. KP showed irregular high reflective white spots in the corneal endothelial microscope images, surrounded by dark areas, and existed in all the eyes with dusty KP found in slit lamp examination and a small number of eyes without obvious KP. Comparing the corneal endothelial cell parameters between the experimental group and the control group, it was found that the corneal endothelial CD and PHC of the former were lower than those of the latter, and the difference was statistically significant (P < 0.001 and P = 0.018, respectively). The AVE and CA of the former were higher than those of the latter (P = 0.013 and P = 0.046, respectively). The corneal endothelial cell density of the eyes with a course of the disease of more than 1 year was lower than that of the eyes with a course of the disease less than 1 year, the coefficient of variation of the corneal endothelial cell area of the eyes with KP was higher than that of the eyes without KP, and the difference was statistically significant (P = 0.003 and P = 0.030, respectively). CONCLUSION: Corneal endothelial microscopy is one of the important methods for the detection of uveitis with high sensitivity. The change of morphological parameters of corneal endothelial cells is one of the important indexes to assist in the diagnosis of uveitis and can be further promoted in ophthalmological examination.

[VEGF short hairpin RNA expression plasmid blocks VEGF expression in rat vascular endothelial cell and inhibits rat corneal neovascularization].

OBJECTIVE: To investigate RNA interference in rat corneal neovascularization and vascular endothelial cells of vascular endothelial growth factor expression and its mechanism. METHODS: In empirical study short hairpin RNA (shRNA) targeting VEGF mRNA was designed and synthesized and VEGF shRNA eukaryotic plasmids were transfected into EC with Lipofectamine 2000 transfection system. Expression of VEGF mRNA and protein in transfected cells was detected by RT-PCR and Western blot. Twenty SD rats were randomized into two groups by randomly Design, experimental group were 10 and 10 in the control group. CNV was induced in vivo by alkaline cauterization of the central cornea, the rats received subconjunctival injection of VEGF shRNA3 plasmid. Then corneal neovascularization was evaluated with microscope. After seven days, VEGF mRNA and protein was determined by RT-PCR and Western blot respectively. RESULTS: VEGF shRNA expression plasmids were successfully constructed and identified by restriction enzyme and sequence analysis. VEGF expression was down regulated by VEGF shRNA1, 2, 3 plasmids, and VEGF shRNA3 plasmid showed stronger inhibitory effect. The inhibitory effects of VEGF mRNA and protein are 68.92% and 66.22%. At the 3, 5 and 7 days after alkali cauterization, the CNV length of the experimental group was obviously decreased comparing with the control group, the difference between two groups has statistics meaning (F=402.700, P=0.000); the CNV area of the experimental group was also obviously decreased comparing with the control group, the difference between two groups has statistics meaning (F=402.700, P=0.000); the VEGF mRNA and protein in corneas was also decreased, the inhibitory effects of which are 41.84% and 41.86%. CONCLUSIONS: RNAi significantly suppresses expression of VEGF in rat EC in vitro. In vivo, subconjunctival delivery of VEGF shRNA3 plasmid suppresses injury-induced VEGF expression and CNV.