ABSTRACT
AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.
ABSTRACT
<p><b>BACKGROUND</b>Although clinical treatment for heart failure and sudden death has been improved over the last few decades, the morbidity and mortality of dilated cardiomyopathy (DCM) have increased. So a better understanding of the underlying molecular events leading to DCM is urgent. Persistent viral infection (especially coxsackievirus group B3) of the myocardium in viral myocarditis and DCM has never been neglected by experts. Recent data indicate that the up-regulation of coxsackievirus and adenovirus receptor (CAR) in viral cardiomyopathy contributes to viral infection as a key factor in the pathogenesis of this disease. This study aimed to investigate the role and regulatory mechanism of CAR in DCM by the bioinformatic method.</p><p><b>METHODS</b>We identified the clusters of genes co-expressed with CAR by clustering algorithm based on the public available microarray dataset of DCM (Kittleson, et al. 2005), and mapped these genes into the protein-protein interaction networks to investigate the interaction relationship to each other at the protein level after confirming that the samples are characterized by the cluster of genes in correctly partitioning.</p><p><b>RESULTS</b>The gene cluster GENESET 11 containing 33 genes including CAR with similar expression pattern was identified by cluster algorithm, of which 19 genes were found to have interaction information of the protein encoded by them in the current human protein interaction database. Especially, 12 genes present as critical nodes (called HUB node) at the protein level are involved in energy metabolism, signal transduction, viral infection, immuno-response, cell apoptosis, cell proliferation, tissue repair, etc.</p><p><b>CONCLUSIONS</b>The genes in GENESET 11 together with CAR may play a pathogenic role in the development of DCM, mainly involved in the mechanism of energy metabolism, signal transduction, viral infection, immuno-response, cell apoptosis and tissue repair.</p>
Subject(s)
Humans , Cardiomyopathy, Dilated , Computational Biology , Coxsackie and Adenovirus Receptor-Like Membrane Protein , Multigene Family , Receptors, Virus , Genetics , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the genetic pathogenesis of dilated cardiomyopathy (DCM) by examining the heterogeneity of Coxsackievirus binding domain (exon 4) of Adenovirus receptor (CAR) in DCM patients and healthy adults, and to detect possible mutation site in exon 4.</p><p><b>METHODS</b>Using polymerase chain reaction (PCR), we amplified exon 4 of CAR DNA extracted from blood samples obtained from 50 DCM patients and 40 healthy adults. The PCR products were screened with single-strand conformation polymorphism (SSCP) and then sequenced alternatively based on the SSCP results.</p><p><b>RESULTS</b>The segment of CAR exon 4 was successfully amplified and there was no single strand conformational disparity in all samples examined by SSCP. Sequence analysis demonstrated that all amplified sequences of CAR exon 4 from samples of the two groups were identical and there was no genetic heterogeneity of CAR exon 4 between the two groups.</p><p><b>CONCLUSION</b>The genetic heterogeneity of CAR exon 4 might not be responsible for the pathogenesis of DCM.</p>