ABSTRACT
Background: Assessment and validation of endothelial-mesenchymal transition (EndoMT) in the retinal endothelium of patients with proliferative diabetic retinopathy (PDR) at the level of retinal and vitreous specimens, and preliminary discussion of its regulatory mechanisms. Methods: Transcriptome sequencing profiles of CD31+ cells from 9 retinal fibrovascular mem-branes (FVMs) and 4 postmortem retinas were downloaded from GEO databases to analyze EndoMT-related differentially expressed genes (DEGs). Then, 42 PDR patients and 34 idiopathic macular holes (IMH) patients were enrolled as the PDR and control groups, respectively. Vitreous humor (VH) samples were collected, and the expression of EndoMT-related proteins was quantified by enzyme-linked immunosorbent assay. Results: A total of 5845 DEGs were identified, and we subsequently focused on the analysis of 24 EndoMT-related marker genes, including the trigger of EndoMT, endothelial genes, mesenchymal genes, transcription factors, inflammatory factors, and autophagy markers. Six of these genes were selected for protein assay validation in VH, showing increased mesenchymal marker (type I collagen α 2 chain, COL1A2) and decreased endothelial marker (VE-cadherin, CDH5) accompanied by increased TGFß, IL-1ß, LC3B and P62 in PDR patients. In addition, anti-VEGF therapy could enhance EndoMT-related phenotypes. Conclusions: EndoMT may underlie the pathogenesis of PDR, and the induction and regulation correlate with autophagy defects and the inflammatory response.
ABSTRACT
PURPOSE: An ovariectomized mouse model was constructed to observe the dynamic effects of hormone changes on the expression of interleukin (IL)-17A and IL-23 in the lacrimal glands. METHODS: The ovariectomized mouse model was constructed by bilateral ovary removal. The concentrations of serum estradiol and testosterone in mouse cardiac blood were detected by radioimmunoassay. Mice in both groups underwent the phenol red cotton thread test and corneal fluorescein staining to assess the ocular surface, whereas Th17 cells in blood and spleen were detected by flow cytometry. IL-17A and IL-23 expression in the lacrimal glands was detected by immunohistochemistry, enzyme-linked immunosorbent assay, and real-time fluorescence quantitative polymerase chain reaction. RESULTS: Serum estradiol and testosterone levels were significantly lower in the ovariectomized group compared with those in the control group. There was lymphocytic infiltration in the lacrimal gland of the ovariectomized group mice. At 6 months after the surgery, aqueous tear production was significantly lower, and statistically significant corneal fluorescein staining was found in the ovariectomized group, compared with that in the control group. In the ovariectomized group, IL-17A and the IL-23 expression in the lacrimal glands and the Th17 expression in the blood and spleen were significantly higher than in the control group. CONCLUSION: The hormone levels are significantly reduced and lymphocytic infiltration in the lacrimal gland in ovariectomized mice, whereas the frequency of Th17 cells in the blood and spleen and IL-17A and IL-23 expression in the lacrimal glands are increased, leading to reduced tear production and positive fluorescein staining in the cornea.
Subject(s)
DNA/genetics , Dry Eye Syndromes/genetics , Gene Expression Regulation , Interleukin-17/genetics , Interleukin-23/genetics , Lacrimal Apparatus/metabolism , Tears/chemistry , Animals , Cornea/metabolism , Cornea/pathology , Disease Models, Animal , Dry Eye Syndromes/metabolism , Dry Eye Syndromes/pathology , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Immunohistochemistry , Interleukin-17/biosynthesis , Interleukin-23/biosynthesis , Lacrimal Apparatus/pathology , Mice , Ovariectomy , Real-Time Polymerase Chain Reaction , Th17 Cells/metabolism , Th17 Cells/pathologyABSTRACT
OBJECTIVES: This study aimed to analyze the influencing factors of meibomian gland loss in patients with ocular chronic graft-versus-host disease (GVHD) and its correlation with other clinical ocular surface assessment indexes. METHODS: The clinical data of patients with ocular chronic GVHD from the ocular surface and corneal outpatient clinic of the First Affiliated Hospital of Soochow University were reviewed from June 2012 to June 2016. The relationship between the percentage of meibomian gland acinar area and the influencing factors, such as patient's sex, age, transplantation method, human leukocyte antigen (HLA) matching, the time when eye discomfort appeared after transplantation (month), and the severity of ocular chronic GVHD, was investigated. Moreover, the correlation between the percentage of meibomian gland acinar area and other ocular surface assessment indexes, such as subjective ocular symptom score, eyelid margin score, tear film break-up time, corneal and conjunctival staining score, and the Schirmer test, was analyzed. RESULTS: A linear relationship was found between the percentage of meibomian gland acinar area and the severity of ocular chronic GVHD, but no linear relationship existed between the percentage of meibomian gland acinar area and the factors such as sex, age, transplantation method, HLA matching, and the time when eye discomfort appeared after transplantation. A negative correlation was observed between the percentage of meibomian gland acinar area and eyelid margin score. CONCLUSIONS: The percentage of meibomian gland acinar area can reflect the severity of ocular chronic GVHD. The clinical significance of the percentage of meibomian gland acinar area needs further investigation.