ABSTRACT
The underlying etiology of diabetic microvascular disease remains unknown. To examine the potential contribution of basic fibroblast growth factor (bFGF), which is an angiogenic factor, and insulin-like growth factor-I (IGF-I) to the development of diabetic microvascular disease, bFGF and IGF-I mRNA levels were measured in tissues of control, diabetic, and insulin-treated diabetic rats. Diabetes was induced in rats by intravenous injection of streptozotocin (STZ) 65 mg/kg, and the rats were maintained for 21 days. bFGF mRNA levels increased threefold in the eyes of diabetic versus control rats, whereas a consistent change in bFGF mRNA levels was not observed in other tissues. In contrast, IGF-I mRNA levels decreased in the eyes and other tissues, including kidney, lung, and skeletal muscle, of diabetic as compared with control rats. Insulin treatment prevented the diabetes-induced increase in bFGF and decrease in IGF-I mRNA levels. Acidic FGF (aFGF) mRNA levels were unchanged in eyes from diabetic versus control rats. In partially purified retinas, diabetes increased bFGF mRNA levels twofold as compared with levels in control retinas, whereas IGF-I mRNA levels decreased to 58% of control levels in retinas from diabetic rats. Insulin treatment again prevented the diabetes-induced increase in IGF-I mRNA levels in the retina but had no effect on the diabetes-induced increase in bFGF mRNA levels. bFGF peptide levels were minimally increased in diabetic versus control retinas.(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Diabetes Mellitus, Experimental/metabolism , Eye/chemistry , Fibroblast Growth Factor 2/genetics , Imidazolidines , Insulin-Like Growth Factor I/genetics , RNA, Messenger/analysis , Aldehyde Reductase/antagonists & inhibitors , Animals , Diabetes Mellitus, Experimental/complications , Diabetes Mellitus, Experimental/genetics , Diabetic Angiopathies/etiology , Diabetic Angiopathies/physiopathology , Diabetic Retinopathy/etiology , Diabetic Retinopathy/physiopathology , Eye/metabolism , Fibroblast Growth Factor 2/analysis , Fibroblast Growth Factor 2/metabolism , Fructose/analysis , Fructose/metabolism , Glucose/analysis , Glucose/metabolism , Imidazoles/pharmacology , Inositol/analysis , Inositol/metabolism , Insulin-Like Growth Factor I/analysis , Insulin-Like Growth Factor I/metabolism , Kidney/chemistry , Kidney/metabolism , Lens, Crystalline/chemistry , Lens, Crystalline/metabolism , Lung/chemistry , Lung/metabolism , Male , Muscle, Skeletal/chemistry , Muscle, Skeletal/metabolism , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Sorbitol/pharmacology , StreptozocinABSTRACT
Insulin-like growth factor-I (IGF-I) is involved in repair and regeneration in tissues in which non-GH-mediated regulation of its production has been shown to be important. We have investigated the effects of a second messenger signaling pathway, intracellular calcium, on IGF-I mRNA levels in cultured rat dermal fibroblasts using a RNase protection assay. Intracellular calcium concentrations ([Ca2+]i) were increased using either the calcium ionophore A23187 or thapsigargin. The ability of these agents to increase [Ca2+]i was confirmed by spectrofluorimetry, using fluo-3 as the Ca2+ indicator. Treatment of cells in serum-free medium and 0.25% BSA [Minimum Essential Medium (MEM) + BSA] with 500 nM A23187 or 1 micron thapsigargin decreased IGF-I mRNA levels in a time-responsive manner over 4-8 h. A23187 and thapsigargin also decreased IGF-I mRNA levels to 36% and 47% of control levels, respectively, in a dose-responsive fashion. Basic fibroblast growth factor mRNA levels, which were simultaneously determined, were either unchanged or increased in cells treated with thapsigargin or A23187. Consistent with the change in IGF-I mRNA levels, immunoreactive IGF-I levels in medium conditioned for 48 h by A23187 or thapsigargin decreased to 25% and 14%, respectively, of control levels in cells maintained in MEM + BSA. To determine the role of protein synthesis in the effects of A23187 and thapsigargin, cells were treated with these agents in the presence or absence of cycloheximide. Cycloheximide had no effect on the decrease in IGF-I mRNA levels mediated by thapsigargin, but significantly attenuated the response to A23187. Given these differences in the role of protein synthesis in and the time course of the effects of A23187 and thapsigargin on IGF-I mRNA levels, additivity experiments were performed. Treatment of cells with the combination of A23187 and thapsigargin resulted in IGF-I mRNA levels that were approximately 70% of the levels present in cells treated with either agent alone. These data are consistent with a small additive effect, but suggest that the majority of the effect of A23187 and thapsigargin occurs via the same final pathway.(ABSTRACT TRUNCATED AT 400 WORDS)
