Source:http://linkedlifedata.com/resource/pubmed/id/10361857
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
6
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| pubmed:dateCreated |
1999-8-3
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| pubmed:abstractText |
In diabetes mellitus, enhanced activity of mesangial cell protein kinase C (PKC) may contribute to nephropathy. The purpose of this study was to determine whether high glucose alters mesangial cell diacylglycerol-sensitive PKC-alpha, -beta2, -delta, and -epsilon content, cellular distribution, and activity through polyol pathway activation. Primary cultured rat mesangial cells (passage 10) were growth-arrested in 0.5% fetal bovine serum and cultured in 5.6 mM glucose (NG) or 30 mM glucose (HG) for 48 h, with or without the aldose reductase inhibitor tolrestat or ARI-509. PKC isoform content in total cell lysates, or cytosol, membrane (Triton X-soluble), and particulate (sodium dodecyl sulfate-soluble) fractions was analyzed by immunoblotting, and band density in HG was expressed as a percentage of corresponding NG values. In HG at 48 h, increased total PKC-alpha (222 +/- 17% of NG, P < 0.001), -beta2 (209 +/- 12%, P < 0.001), and -epsilon (195 +/- 19%, P < 0.001) were observed. L-Glucose had no effect on total PKC isoform content. HG caused increased membrane- and particulate-associated PKC-alpha (257 +/- 87 and 327 +/- 66%, respectively, P < 0.05), membrane-associated PKC-delta (143 +/- 10%, P < 0.05), and membrane-associated PKC-epsilon (186 +/- 11%, P < 0.001), with no change in cytosol contents. The HG effects were not mimicked by L-glucose. In NG or HG, PKC-beta2 was not detected in the cytosol fraction, and membrane and particulate association were unchanged with phorbol ester stimulation. Confocal immunofluorescence imaging revealed that in HG, PKC-alpha, -delta, and -epsilon translocate to the nucleus and plasma membrane. Total PKC activity measured by in situ 32P-phosphorylation of the epidermal growth factor receptor substrate increased from 18 +/- 1 pmol/min per mg cell protein in NG to 33 +/- 3 pmol/min per mg cell protein in HG (P < 0.002 versus NG). In NG, tolrestat and ARI-509 exposure caused increased PKC activity, enhanced accumulation of total PKC-alpha and -beta2, with no change in total or fractional recovery of PKC-delta or -epsilon. In HG, tolrestat and ARI-509 prevented the increase in total PKC-epsilon and membrane-associated PKC-delta and -epsilon. It is concluded that within 48 h of HG, enhanced mesangial cell PKC activity is associated with accumulation and cellular redistribution of diacylglycerol-sensitive PKC isoforms, and that increased PKC-epsilon content and membrane-associated PKC-delta and -epsilon are dependent on polyol pathway activation.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Enzyme Inhibitors,
http://linkedlifedata.com/resource/pubmed/chemical/Glucose,
http://linkedlifedata.com/resource/pubmed/chemical/Isoenzymes,
http://linkedlifedata.com/resource/pubmed/chemical/Naphthalenes,
http://linkedlifedata.com/resource/pubmed/chemical/Polymers,
http://linkedlifedata.com/resource/pubmed/chemical/Prkcd protein, rat,
http://linkedlifedata.com/resource/pubmed/chemical/Prkce protein, rat,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Kinase C,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Kinase C-delta,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Kinase C-epsilon,
http://linkedlifedata.com/resource/pubmed/chemical/Sorbitol,
http://linkedlifedata.com/resource/pubmed/chemical/polyol,
http://linkedlifedata.com/resource/pubmed/chemical/tolrestat
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| pubmed:status |
MEDLINE
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| pubmed:month |
Jun
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| pubmed:issn |
1046-6673
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:volume |
10
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
1193-203
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| pubmed:dateRevised |
2009-11-19
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| pubmed:meshHeading |
pubmed-meshheading:10361857-Animals,
pubmed-meshheading:10361857-Blotting, Western,
pubmed-meshheading:10361857-Cattle,
pubmed-meshheading:10361857-Cells, Cultured,
pubmed-meshheading:10361857-Dose-Response Relationship, Drug,
pubmed-meshheading:10361857-Drug Interactions,
pubmed-meshheading:10361857-Enzyme Inhibitors,
pubmed-meshheading:10361857-Glomerular Mesangium,
pubmed-meshheading:10361857-Glucose,
pubmed-meshheading:10361857-Isoenzymes,
pubmed-meshheading:10361857-Male,
pubmed-meshheading:10361857-Microscopy, Confocal,
pubmed-meshheading:10361857-Naphthalenes,
pubmed-meshheading:10361857-Polymers,
pubmed-meshheading:10361857-Protein Kinase C,
pubmed-meshheading:10361857-Protein Kinase C-delta,
pubmed-meshheading:10361857-Protein Kinase C-epsilon,
pubmed-meshheading:10361857-Rats,
pubmed-meshheading:10361857-Rats, Sprague-Dawley,
pubmed-meshheading:10361857-Reference Values,
pubmed-meshheading:10361857-Sensitivity and Specificity,
pubmed-meshheading:10361857-Sorbitol
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| pubmed:year |
1999
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| pubmed:articleTitle |
Effect of high glucose on mesangial cell protein kinase C-delta and -epsilon is polyol pathway-dependent.
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| pubmed:affiliation |
Institute of Medical Science and Department of Medicine, University of Toronto, Ontario, Canada.
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| pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, Non-U.S. Gov't
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