Determination of sorbitol and galactitol at the nanogram level in biological samples by high-performance liquid chromatography

Dethy, Jean-Marie; Callaert-Deveen, Bethy; Janssens, Martine; Lenaers, A.

doi:10.1016/0003-2697(84)90565-7

Cited by 42 publications

(14 citation statements)

References 13 publications

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“…Supernatants were deproteinized with the Centrifree Micropartition System (Amicon Corp., Danvers, MA) and fractionated on a Sugar Pak column (6.5 mm X 30 cm; Waters Associates, Milford, MA). The inositol fraction was then derivatized with p-chloronitrobenzene and further fractionated on a Waters microporasil column (6.5 mm X 30 cm) as previously described (24). The p-chloronitrobenzoate derivative of inositol was quantitated by absorption at 254 Mm.…”

Section: Methodsmentioning

confidence: 99%

Protein kinase C is activated in glomeruli from streptozotocin diabetic rats. Possible mediation by glucose.

Craven¹,

DeRubertis²

1989

J. Clin. Invest.

321

170

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Glomerular inositol content and the turnover of polyphosphoinositides was reduced by 58% in 1-2 wk streptozotocin diabetic rats. Addition of inositol to the incubation medium increased polyphosphoinositide turnover in glomeruli from diabetic rats to control values. Despite the reduction in inositol content and polyphosphoinositide turnover, protein kinase C was activated in glomeruli from diabetic rats, as assessed by an increase in the percentage of enzyme activity associated with the particulate cell fraction. Total protein kinase C activity was not different between glomeruli from control and diabetic rats. Treatment of diabetic rats with insulin to achieve near euglycemia prevented the increase in particulate protein kinase C. Moreover, incubation of glomeruli from control rats with glucose (100-1,000 mg/dl) resulted in a progressive increase in labeled diacylglycerol production and in the percentage of protein kinase C activity which was associated with the particulate fraction. These results support a role for hyperglycemia per se in the enhanced state of activation of protein kinase C seen in glomeruli from diabetic rats. Glucose did not appear to increase diacylglycerol by stimulating inositol phospholipid hydrolysis in glomeruli. Other pathways for diacylglycerol production, including de novo synthesis and phospholipase C mediated hydrolysis of phosphatidylcholine or phosphatidylinositol-glycan are not excluded.

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Section: Methodsmentioning

confidence: 99%

Protein kinase C is activated in glomeruli from streptozotocin diabetic rats. Possible mediation by glucose.

Craven¹,

DeRubertis²

1989

J. Clin. Invest.

321

170

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show abstract

“…3 N barium hydroxide and centrifuged at 10 000 g for 10 min. A 1 ml aliquot of supernatant was lyophilized overnight, and the lenticular sorbitol levels were measured by highperformance liquid chromatography (HPLC) using a modi®cation of the method of Dethy et al (1984).…”

Section: Biochemical Analysismentioning

confidence: 99%

Biochemical and Morphological Changes during Development of Sugar Cataract in Otsuka Long–Evans Tokushima Fatty (OLETF) Rat

Kubo

Maekawa

Tanimoto

et al. 2001

Experimental Eye Research

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“…Cells were washed with cold saline several times, homogenized in 2 ml cold water with the internal standard, deproteinized with ethanol to a final concentration of 70%, and centrifuged at 12,000 x g for 15 min. The supernatant was lyophilized and derivatized with phenyl isocyanate as previously reported by Dethy et al (20). Polyol derivatives were then applied to a column (Nucleosil 5C 18 MN, Machery-Nagel, Duren, FRG), eluted isocratically with 0.01 M phosphate buffer (pH 7.4) containing 60% acetonitrile at a flow rate of 0.8 ml/min, and detected at 240 nm.…”

Section: Methodsmentioning

confidence: 99%

Evidence for Existence of Polyol Pathway in Cultured Rat Mesangial Cells

et al. 1987

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SUMMARYThe accumulation of polyols has been previously found in renal glomeruli isolated from streptozocin-induced diabetic (STZ-D) rats, although the intraglomerular polyol pathway has not been exactly localized. Because we have previously observed mesangial cell dysfunction in STZ-D rats, we examined whether the polyol pathway exists in mesangial cells as a possible candidate of the cause of cellular dysfunction. The activities of two polyol pathway enzymes, aldose reductase and sorbitol dehydrogenase, were clearly detected in the crude homogenate of cultured mesangial cells at higher levels than those of whole glomeruli when DL-glyceraldehyde or D-fructose was used as substrate. When cells were incubated in medium containing 55 mM glucose or galactose, a large amount of sorbitol or galactitol was accumulated intracellularly. The accumulation of polyols was effectively blocked by an aldose reductase inhibitor, ICI 128436. These results suggest that the polyol pathway exists in mesangial cells of rat glomeruli and may play a role in the development of mesangial cell dysfunction found in STZ-D rats. Diabetes 36:240-43, 1987 T here has been increasing evidence that the enhanced activity of polyol pathway plays an important role in the development of various diabetic complications such as cataract, neuropathy, and retinopathy (1,2). Furthermore, recent reports by indicate that diabetic nephropathy might also be caused by sorbitol accumulation in glomeruli. Because the activity of aldose reductase, a key enzyme for sorbitol synthesis, was found immunohistochemically in glomerular epithelium (6), sorbitol accumulation in epithelial cells may be pathogenetically linked to the development of diabetic nephropathy.On the other hand, it has been generally believed that the abnormality of mesangial region plays a crucial role in the development and progression of diabetic glomerulosclerosis (7). Mesangial expansion is known to cause the obliteration of glomerular capillary lumen (8) and to correlate well with clinical parameters of renal involvement in diabetic patients (9). Although the precise mechanism of the development of mesangial expansion has not yet been clarified, the derangement of mesangial cells may be one of the etiological factors (10,11). Therefore, it appears pertinent to investigate whether the activity of polyol pathway exists in mesangial cells in connection with mesangial abnormality. MATERIALS AND METHODSGlomerular isolation and culture. Glomeruli were isolated from male Sprague-Dawley rats weighing 50-100 g by sieving with stainless steel and nylon meshes under sterile conditions as previously reported (12). The yield of glomerular isolation by this technique was -4 0 % and the purity was >90% and was comparable to results obtained in human autopsied kidney using similar technique (13,14). Isolated glomeruli were then cultured with RPMI-1640 culture medium (Gibco, Grand Island, NY) with 20% fetal calf serum plus penicillin and streptomycin according to the method of Kreisberg and Karnovsky (15)....

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Determination of sorbitol and galactitol at the nanogram level in biological samples by high-performance liquid chromatography

Cited by 42 publications

References 13 publications

Protein kinase C is activated in glomeruli from streptozotocin diabetic rats. Possible mediation by glucose.

Protein kinase C is activated in glomeruli from streptozotocin diabetic rats. Possible mediation by glucose.

Biochemical and Morphological Changes during Development of Sugar Cataract in Otsuka Long–Evans Tokushima Fatty (OLETF) Rat

Evidence for Existence of Polyol Pathway in Cultured Rat Mesangial Cells

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