Attenuation of phosphoinositidase activity and phosphatidylinositol bisphosphate level of bovine retinal capillary pericytes in high glucose.
Both phosphoinositidase (PIase) and individual species of inositol phospholipid (IPL) of bovine retinal capillary pericytes (BRCP) were quantitatively determined. When glucose in growth medium was increased from 5- to 15- or 30 mM, PIase activity was attenuated to 82% or 55%, respectively. In contrast, when glucose (5-, 15-, 30 mM) was added to an enzyme extract from cells grown in the standard growth medium (5 mM glucose, 0.04 mM myo-inositol) the PIase activity was not changed, indicating that the reduced PIase activity was not due to the direct effect of glucose. When IPLs from BRCP were analysed by HPLC and TLC, we observed reduction of the total and newly formed IPLs including the substrate of PIase. Phosphatidylinositol bisphosphate (PIP2). Reduced levels of IPLs were associated with a decrease in myo-inositol and an increase in sorbitol. The changes in IPL metabolism were reversed by adding either free myo-inositol or AL1576, an aldose reductase inhibitor (ARI), to the high-glucose medium. However, the addition of myo-inositol to the growth medium with a standard concentration of glucose only caused a marked increase in phosphatidylinositol, but not in PIP or PIP2, while the supplement of AL1576 in the standard medium did not cause any changes in IPL formation. These findings suggest that the alteration in IPL metabolism in BRCP may be related to insufficient myo-inositol or activated sorbitol pathway under high-glucose conditions. Further explanation of the role of the altered hydrolysis of PIP2 triggered by PIase may provide clues to understanding of the mechanism of decreased pericyte viability in the presence of high glucose concentrations.
Li
W Y
WY
Department of Ophthalmology, Peking Union Medical College Hospital, Beijing, People's Republic of China.
Tang
L
L
Zhou
Q
Q
Qin
M
M
Hu
T S
TS
D019870:1-Phosphatidylinositol 4-Kinase; D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors; D000818:Animals; D002196:Capillaries / Q000201:enzymology; D002417:Cattle; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005449:Fluorenes / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology*; D006827:Hydantoins / Q000494:pharmacology; D007294:Inositol / Q000494:pharmacology; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism*; D012171:Retinal Vessels / Q000201:enzymology*; D013012:Sorbitol / Q000494:pharmacology
D005449:Fluorenes; D006827:Hydantoins; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; C037178:phosphatidylinositol 4-phosphate; C052241:imirestat; D007294:Inositol; D013012:Sorbitol; D000449:Aldehyde Reductase; D010770:Phosphotransferases; D019870:1-Phosphatidylinositol 4-Kinase; D005947:Glucose