| Partâ… Screening of differentially expressed genes in the jejunum of rats with idiopathic hyperoxaluriaObjecttive Idiopathic hyperoxaluria may be caused by increased endogenous formation or exogenous absorption of oxalic acid. Characterization of the molecular pathogenesis of idiopathic hyperoxaluria has been hampered by the lack of an ideal animal model. We therefore established a stable idiopathic hyperoxaluria rat model in order to analyze variation in gene expression profiling in the jejunum and to investigate the association between idiopathic hyperoxaluria pathogenesis and exogenous absorption of oxalic acid.Methods A rat model of idiopathic hyperoxaluria was established and three female rats with idiopathic hyperoxaluria were assigned to the study group, while three normal rats served as controls. Total RNA was isolated from the jejunum of rats in the two groups and mRNA was purified, reversely transcribed, labeled with Cy5 or Cy3 and hybridized to 27K Rat Genome Array. Differences in gene expression profile between the 2 groups were analyzed by bioinformatics methods.Results Comparative analysis showed that 517 genes expression was up-regulated and 203 genes expression was down-regulated by at least two-fold in the jejunum of rats with idiopathic hyperoxaluria. These genes are related to many functions including DNA binding and transcription, cell signal transduction, ATP binding, ion binding and transport, cell receptors, immunity, cyclins, cytoskeleton structure, and metabolic proteins, among others. KEGG (Kyoto encyclopedia of genes and genomes) signaling pathway analysis revealed that the variations of 239 pathway functional changes are statistically significant (P<0.05).Conclusions cDNA microarray can be used effectively to screen differentially expressed genes of idiopathic hyperoxaluria rats in the jejunum. These differentially expressed genes may underlie idiopathic hyperoxaluria pathophysiology and provide a platform for further studying molecular pathogenetic mechanisms. Partâ…¡Feasibility study of jejunum protein in rats with idiopathic hyperoxaluria by two-dimensional gel electrophoresisObjective To evaluate the feasibility of the jejunum protein in rats with idiopathic hyperoxaluria by two-dimensional gel electrophoresis(2-DE),and to lay experimental foundation for official 2-DE.Methods One female rat with idiopathic hyperoxaluria was as study group, and one normal femal rat was as control. The jejunum tissue of idiopathic hyperoxaluria rat and the normal rat were homogenized with lysis solution. Samples of saliva protein were separated and the electrophoregrams were established by 2-DE.The electrophoregrams were analyzed by ImageMasterTM 2D Platinum Version 5.0 software for detecting the protein spot.Results Satisfying two-dimensional electrophoregrams of jejunum protein in rats were obtained.2654 protein spots were detected in rat with idiopathic hyperoxaluria while 2541 in normal ones.Conclusions The mobility shift of the protein spots was stable in the two electrophoregrams, which ensure the stable repeatability and feasibility of official 2-DE in future studies. Partâ…¢Identification of differentially expressed proteins in the jejunum in a rat model with idiopathic hyperoxaluriaObjective To characterize differentially expressed proteins in the jejunum in a newly established rat model of idiopathic hyperoxaluria, and to analyze the pathogenesis of IH caused by exogenous oxalate.Methods A rat model of idiopathic hyperoxaluria was established and 3 female rats of idiopathic hyperoxaluria were designed to the study group, and 3 normal rats served as controls. Protein was extracted from each rat's jejunum and separated by two-dimensional electrophoresis. Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was used to identify differentially expressed proteins. Functions and cellular localization of these proteins were determined by gene ontology.Results Based on gel protein profiles which were satisfactory in resolution and reproducibility,40 differentially expressed protein spots,31 up-regulated and 9 down-regulated, were apparent in idiopathic hyperoxaluria rats,34 of them (25 up-regulated including Ppplr8, Psmal, Krt19, Hspb1, et al, and 9 down-regulated including Actb, Krt8, Psma5, et al.) were successfully identified by MALDI-TOF-MS. Most of the differentially expressed proteins have functions related to the cytoskeleton, signal transduction, rotein degradation and others.Conclusions These results unveil that these differentially expressed proteins may be key proteins in idiopathic hyperoxaluria pathophysiology and provide a platform for further studying molecular pathogenetic mechanisms. |
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