The Transcriptive Regulation Of Cultured Cells In Saccharometabolic Abnormality

Objective: To explore the glycobiology-related pathogenesis of diabetic retinopathy by studying the transcriptive regulation of stringently cultured cells responsive to the changes of basic components involved in the process of glucose metabolism.Methods: To set up a stable glucose-free culturing environment for human diploid fibroblast and a few kinds of vascular endothelia for the single-factor analysis. To observe the responsive biological behaviors of the cells after the glucose or glucose-derived molecules content of the culture medium was changed precisely following the stimulation of glucose transport system by insulin. To construct a differentially-expressed cDNA library between any two samples by suppression subtractive hybridization technology.Results: The human fibroblast and all mentioned endothelia could be cultured as usual in glucose-free medium without compromise for necessary time to evaluate the stimulating effect of the exact concentration of glucose and its derivates. Comparing to controls, glucose in medium as high concentration as 20mM did harm to all kinds of cells observed here, and the successfully cloned differentially-expressed cDNA implicated the changes on transcriptive level in HUVEC after 4hr. 2mM GlcN-6-P or GlcNAc-6-P damaged the human fibroblast badly in 4hr while the endothelia shown more resistance.Conclusion: The toxicity of high concentration of glucose or its active derivates to cultured cells might be related to the regulation on transcriptive level. The products of hexosamine biosynthesis pathway are probably potent stimulator during the genesis of diabetic retinopathy. The transcriptive response of vascular endothelium to toxical metabolites could be the key point of the pathogenesis of diabetic retinopathy.