ABSTRACT
We previously demonstrated vasoactive intestinal polypeptide (VIP) eyedrops reduce intraocular pressure (IOP) and stabilize cytoskeleton of the Schlemm's canal (SC) endothelium in a chronic ocular hypertension rat model. Here we determine if the trabecular meshwork (TM) releases endogenous VIP and affect SC in paracrine manner, and whether this cellular interaction via VIP is strengthened under stimulated sympathetic activity. A rat model of moderate-intensity exercise was established to stimulate sympathetic activation. IOP post exercise was measured by a rebound tonometer. Sympathetic nerve activity at the TM was immunofluorescence-stained with DßH and PGP9.5. Morphological changes of TM and SC were quantitatively measured by hematoxylin-eosin (HE) staining. Further, epinephrine was applied to mimic sympathetic excitation on primary rat TM cells, and ELISA to measure VIP levels in the medium. The cytoskeleton protective effect of VIP in the epinephrine-stimulated conditioned medium (Epi-CM) was evaluated in oxidative stressed human umbilical vein endothelial cells (HUVECs). Elevated sympathetic nerve activity was found at TM post exercise. Changes accompanying the sympathetic excitation included thinned TM, expanded SC and decreased IOP, which were consistent with epinephrine treatment. Epinephrine decreased TM cell size, enhanced VIP expression and release in the medium in vitro. Epi-CM restored linear F-actin and cell junction integrity in H2O2 treated HUVECs. Blockage of VIP receptor by PG99-465 attenuated the protective capability of Epi-CM. VIP expression was upregulated at TM and the inner wall of SC post exercise in vivo. PG99-465 significantly attenuated exercise-induced SC expansion and IOP reduction. Thus, the sympathetic activation promoted VIP release from TM cells and subsequently expanded SC via stabilizing cytoskeleton, which resulted in IOP reduction.
Subject(s)
Trabecular Meshwork , Vasoactive Intestinal Peptide , Animals , Humans , Rats , Actins/metabolism , Culture Media, Conditioned/pharmacology , Epinephrine/metabolism , Human Umbilical Vein Endothelial Cells , Hydrogen Peroxide/pharmacology , Intraocular Pressure , Ophthalmic Solutions/pharmacology , Receptors, Vasoactive Intestinal Peptide/metabolism , Trabecular Meshwork/metabolism , Vasoactive Intestinal Peptide/pharmacology , Vasoactive Intestinal Peptide/metabolismABSTRACT
Long noncoding RNAs (lncRNAs) are a group of noncoding transcripts of >200 nucleotides. They can act as competing endogenous RNAs (ceRNAs) and suppress microRNA (miRNA) function by preventing them from binding to and interacting with target mRNAs. However, the specific role of the lncRNAassociated ceRNA network in the pathogenesis of glaucoma has not yet been elucidated. To study this, data were downloaded from the Gene Expression Omnibus database (GSE126170), which contained three human trabecular meshwork cell (HTMC) samples treated with 300 µm hydrogen peroxide and three control samples treated with vehicle. Differentially expressed lncRNAs and mRNAs of HTMCs were obtained using the R package limma. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of differentially expressed mRNAs were performed using the R package clusterProfiler. Finally, the ceRNA network was constructed using the mircode, miRDB, miRTarBase and TargetScan databases, and visualized using Cytoscape v3.6.1. The results showed that 70 lncRNAs and 558 mRNAs were identified to be significantly dysregulated (|log2FoldChange| >1 and adjusted P<0.05) in HTMCs under oxidative stress compared to those in HTMCs under control conditions. Moreover, 24 lncRNAs, 24 miRNAs and 40 mRNAs were closely connected, and were part of the ceRNA network. Among these, the expression levels of 19 lncRNAs were upregulated, and those of 5 lncRNAs were downregulated. To conclude, using bioinformatics analysis, the differential expression profiles of lncRNAs were reported and a lncRNAassociated ceRNA network in HTMCs under oxidative stress was constructed. These results may bring to light a new pathological mechanism or a potential therapeutic target for glaucoma.
Subject(s)
Computational Biology , MicroRNAs/genetics , Oxidative Stress , RNA, Long Noncoding/genetics , RNA, Messenger/genetics , Trabecular Meshwork/cytology , Trabecular Meshwork/metabolism , Computational Biology/methods , Databases, Genetic , Gene Expression Profiling , Gene Expression Regulation , Gene Ontology , Gene Regulatory Networks , Humans , TranscriptomeABSTRACT
BACKGROUND The aim of this study was to analyze the long non-coding RNA (lncRNA)-associated competing endogenous RNA (ceRNA) network in human retinal tissues following detachment with proliferative vitreoretinopathy (PVR). MATERIAL AND METHODS Expression data of 19 human detached retinas with PVR and 19 normal retinas from postmortem donors were downloaded from Gene Expression Omnibust (GEO) database (GSE28133). The R package "limma" was utilized to discriminate the dysregulated lncRNA and mRNA profiles. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses of differentially expressed mRNAs were performed using R packages "Clusterprofiler." The ceRNA network of dysregulated genes was constructed by using mircode, miRDB, miRTarBase and TargetScan databases, and was visualized by Cytoscape v3.6.1. RESULTS A total of 23 lncRNAs and 994 mRNAs were identified significantly expressed between the human detached retinas with PVR and the normal retina tissues, with thresholds of |log2FoldChange| >1.0 and adjusted P-value <0.05. The constructed ceRNA network (lncRNA-miRNA-mRNA regulatory axis) included 9 PVR-specific lncRNAs, as well as 27 miRNAs and 73 mRNAs. CONCLUSIONS We demonstrated the diï¬erential lncRNA expression profile and constructed a lncRNA-associated ceRNA network in human detached retinas with PVR. This may ferret out an unknown ceRNA regulatory network in human retinal detachment with PVR.
Subject(s)
RNA, Long Noncoding/genetics , Retinal Detachment/genetics , Vitreoretinopathy, Proliferative/genetics , Computational Biology/methods , Databases, Genetic , Gene Expression Profiling/methods , Gene Expression Regulation, Neoplastic/genetics , Gene Ontology , Gene Regulatory Networks/genetics , Humans , Kaplan-Meier Estimate , MicroRNAs/genetics , RNA, Long Noncoding/analysis , RNA, Messenger/geneticsABSTRACT
BACKGROUND Patients with primary breast cancer following primary ovarian cancer do not comprise a large clinical entity, and reports of the survival outcomes of this cohort are rare. The purpose of this retrospective population-based research was to investigate the survival outcomes of patients with primary breast cancer after primary ovarian cancer. MATERIAL AND METHODS A cohort of patients diagnosed with primary breast cancer following primary ovarian cancer between 1973 and 2014 was drawn from the National Cancer Institute's Surveillance Epidemiology and End Results (SEER) database. Cox proportional hazards survival regression analysis and Kaplan-Meier were applied to calculate overall survival (OS), cancer-specific survival (CSS), and independent predictors of CSS. RESULTS A total of 1455 patients with primary breast cancer following primary ovarian cancer were identified. The 5-year and 10-year OS rates for the entire cohort were 81.7% and 67.4%, respectively. The 5-year and 10-year CSS rates were 84.2% and 74.3% for ovarian cancer, and 76.0% and 67.8% for breast cancer, respectively. Multivariate analysis revealed that independent predictors of ovarian cancer CSS include age, cancer stage, diagnosis time, and histological subtype. CONCLUSIONS Patients diagnosed with breast cancer following ovarian cancer have better survival rates. Patients age, ovarian cancer stage, ovarian cancer histological type, and time of diagnose affect the survival rate.
