Screening Of Differential Expression Genes Between Diabetic Wounds And Normal Skin Wounds By CDNA Microarray Technique

Objective: Analyzing and screening of differential expression genes of Diabeticrats and normal rats wounds by cDNA microarray technique, Preliminary research oncharacteristics of expression genes and their possible biological significance indiabetic rats and normal rats wound, To provide a new way and some new clues forthe treatment of diabetic wounds.Methods: Male SD rats were randomly divided into DM group and normalcontrol group. DM modeled was prepared with a single intraperitoneal injection ofstreptozoocin (STZ). at the dose of65mg/kg. Diameters of2.0cm full-thicknessskin wounds in the back of randomly selected stable molded diabetic rats and normalones were made after3weeks, from which wound tissues were taken72hours later.Total RNA was extracted from above cells by Trizol one-step method，and weredetected by formaldehyde denaturing agarose gel electrophoresis.Cy3fluorescencewas labeled to the cDNA on both groups by reverse method. Probes were preparedand hybridized into cDNA microarray for scanning with Agilent G2565CAMicroarray Scanner and analysis of images. Data extracted by analyzing images withFeature Extraction (v10.7) software were analyzed with Boao microarray AnalyzerSoftware. Differentially expressed genes were screened with two-fold differentialexpression value When image signals were translated into digital signals.Significantlyup/down-regulated genes were selected for verification by real time RT-PCR.Differential expression genes were classified by genes function at the same time.Results: Compared with normal rats group,2206differentially exressed geneswere detected in the group of diabetic rats, including785up-regulated genes and1421down-regulated genes. Then,231genes were significantly up-regulated, and463genes were significantly down-regulated. It was proved that real-time RT-PCRvalidation results was consistent with microarray ones. Given that detectingdefferential genes through the functional classification of genes involved in multiplebiological processes such as protein synthesis, glucose metabolism, ion channels and nerve conduction,Conclusion: Gene expression profiles of the skin wounds in diabetic rats aresignificantly different from that of normal rats are significantly different,, which maybe closely related to the different healing repair ability in skin wound between them.