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... bovine serum at 37°C in an atmosphere of 95% air and 5% CO2. Northern Blot Analysis. Total RNA was isolated by extraction with acid guanidinium thiocyanate-phenol-chloroform as described previously (=-=Chomczynski and Sacchi, 1987-=-) and quantified by measuring the absorbance at 260 nm. Aliquots of 5 g of total RNA, heat-denatured at 65°C for 15 min in gel-running buffer [40 mM MOPS, 10 mM sodium acetate, and 1 mM EDTA, pH 7.0]...

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...s a member of NADPH-dependent aldo-keto reductase family (Jez et al., 1997), has been studied for its involvement in the pathogenesis of various diabetic complications (Kinoshita and Nishimura, 1988; =-=Yabe-Nishimura, 1998-=-). AR catalyzes the reduction of the aldehyde form of glucose to sorbitol, which is subsequently converted to fructose by NAD-dependent sorbitol dehydrogenase. This alternative route of glucose metab...

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...at mediates the induction of the AR gene seemed to differ. ROS transduce their signals via MAP kinases in many cellular processes including proliferation, differentiation, and apoptosis (Davis, 1993; =-=Lopez-Ilasaca, 1998-=-). MG has been reported to induce phosphorylation of not only p38 kinase but also JNK and ERKs in human umbilical vein endothelial cells (Akhand et al., 2001). In Jurkat leukemia T cells, MG also acti...

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...difies cellular proteins to form cross-links of amino groups, generating so-called advanced glycation end products. This modification has been implicated in the development of diabetic complications (=-=Odani et al., 1999-=-). MG is known to have multiple metabolic origins. It is formed by the enzymatic and nonenzymatic elimination of phosphate from triose phosphate (Pompliano et al., 1990) and by oxidation of aminoaceto...

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...c aldehyde MG have been reported during hyperglycemic conditions and have been implicated in the vascular complications of diabetes (Vander Jagt et al., 1992; Yamada et al., 1994; Okado et al., 1996; =-=Che et al., 1997-=-). In the tissues of healthy subjects, the estimated rate of formation of MG was approximately 125 M/day, whereas the concentration of serum MG in patients with diabetes was reported to be elevated u...

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...ing hyperglycemia to elicit impaired ion transport (Greene, 1988), altered redox status (Tilton et al., 1992), deranged osmotic balance (Kinoshita and Nishimura, 1988), and enhanced oxidative stress (=-=Gonzalez et al., 1986-=-). Although numerous AR inhibitors have been developed as new therapeutic agents for diabetic complications, their clinical efficacy has not been proven unequivocally. On the other hand, AR exhibits b...

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...d levels of cytotoxic aldehyde MG have been reported during hyperglycemic conditions and have been implicated in the vascular complications of diabetes (Vander Jagt et al., 1992; Yamada et al., 1994; =-=Okado et al., 1996-=-; Che et al., 1997). In the tissues of healthy subjects, the estimated rate of formation of MG was approximately 125 M/day, whereas the concentration of serum MG in patients with diabetes was reporte...

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... to fructose by NAD-dependent sorbitol dehydrogenase. This alternative route of glucose metabolism, known as the polyol pathway, is accelerated during hyperglycemia to elicit impaired ion transport (=-=Greene, 1988-=-), altered redox status (Tilton et al., 1992), deranged osmotic balance (Kinoshita and Nishimura, 1988), and enhanced oxidative stress (Gonzalez et al., 1986). Although numerous AR inhibitors have bee...

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...s (AGE) induced the augmented expression of AR, leading to the notion of a reciprocal relationship between augmented activity of AR and AGE to promote the development of vascular lesions in diabetes (=-=Nakamura et al., 2000-=-). Long-term exposure to high concentrations of MG may be among the causative factors that accelerate cellular oxidative stress under such pathological conditions as diabetes (Baynes, 1991). Induction...

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...A SPET Journals on M arch 6, 2016 m olpharm .aspetjournals.org D ow nloaded from pathway becomes more significant in other tissues, including the cardiovascular-renal system, in which AR is abundant (=-=Gui et al., 1995-=-; Nishimura et al., 1988; Nishimura et al., 1993; Vander Jagt et al., 2001). Also, a recent report demonstrated that over-expression of the gene encoding AR increases the tolerance against MG in Sacch...

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... enzyme not only for glucose, but also for reactive aldehydes generated in the cells. In fact, the functional significance of AR against cytotoxic aldehydes has been suggested in hepatoma cell lines (=-=Takahashi et al., 1995-=-), in vascular smooth muscle cells (Spycher et al., 1997; Seo et al., 2000; Nishinaka and Yabe-Nishimura, 2001), and in the lesions of vasculitis (Rittner et al., 1999). These findings in conjunction ...

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... and glucose. Elevated levels of cytotoxic aldehyde MG have been reported during hyperglycemic conditions and have been implicated in the vascular complications of diabetes (Vander Jagt et al., 1992; =-=Yamada et al., 1994-=-; Okado et al., 1996; Che et al., 1997). In the tissues of healthy subjects, the estimated rate of formation of MG was approximately 125 M/day, whereas the concentration of serum MG in patients with ...

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...m .aspetjournals.org D ow nloaded from pathway becomes more significant in other tissues, including the cardiovascular-renal system, in which AR is abundant (Gui et al., 1995; Nishimura et al., 1988; =-=Nishimura et al., 1993-=-; Vander Jagt et al., 2001). Also, a recent report demonstrated that over-expression of the gene encoding AR increases the tolerance against MG in Saccharomyces cerevisiae (Aguilera and Prieto, 2001)....

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... M arch 6, 2016 m olpharm .aspetjournals.org D ow nloaded from pathway becomes more significant in other tissues, including the cardiovascular-renal system, in which AR is abundant (Gui et al., 1995; =-=Nishimura et al., 1988-=-; Nishimura et al., 1993; Vander Jagt et al., 2001). Also, a recent report demonstrated that over-expression of the gene encoding AR increases the tolerance against MG in Saccharomyces cerevisiae (Agu...

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...sis. The activity of AR was determined in reaction mixtures containing 0.1 M sodium phosphate buffer, pH 6.2, 150 M NADPH, 10 mM DL-glyceraldehyde, and the enzyme solution in a total volume of 1 ml (=-=Nishimura et al., 1991-=-). The reaction was started by the addition of enzyme, and activity was measured spectrophotometrically by estimating NADPH oxidation from the decrease in absorbance at 340 nm. Assays were carried out...