Glyoxalase I Is Critical for Human Retinal Capillary Pericyte Survival under Hyperglycemic Conditions

• Published in: The Journal of biological chemistry Vol. 281; no. 17; pp. 11864 - 11871
• Main Authors: Miller, Antonia G., Smith, Dawn G., Bhat, Manjunatha, Nagaraj, Ram H.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 28-04-2006; American Society for Biochemistry and Molecular Biology
• Subjects: Adult; Aged; Apoptosis - drug effects; Capillaries - cytology; Capillaries - physiology; Cells, Cultured; Glucose - pharmacology; Humans; Hyperglycemia - metabolism; Hyperglycemia - pathology; Lactoylglutathione Lyase - genetics; Lactoylglutathione Lyase - metabolism; Nitric Oxide - metabolism; Nitric Oxide Donors - pharmacology; Nitroso Compounds - pharmacology; Pericytes - cytology; Pericytes - metabolism; Pyruvaldehyde - metabolism; Retinal Vessels - cytology; Retinal Vessels - physiology; Sweetening Agents - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M513813200

Retinal capillary pericytes undergo premature death, possibly by apoptosis, during the early stages of diabetic retinopathy. The α-oxoaldehyde, methylglyoxal (MGO), has been implicated as a cause of cell damage in diabetes. We have investigated the role of MGO and its metabolizing enzyme, glyoxalase I, in high glucose-induced apoptosis (annexin V binding) of human retinal pericyte (HRP). HRP incubated with high glucose (30 mm d-glucose) for 7 days did not undergo apoptosis despite accumulation of MGO. However, treatment with a combination of high glucose and S-p-bromobenzylglutathione cyclopentyl diester, a competitive inhibitor of glyoxalase I, resulted in apoptosis along with a dramatic increase in MGO. Overexpression of glyoxalase I in HRP protected against S-p-bromobenzylglutathione cyclopentyl diester-induced apoptosis under high glucose conditions. Incubation of HRP with high concentrations of MGO resulted in an increase of apoptosis relative to untreated controls. We found an elevation of nitric oxide (NO·) in HRP that was incubated with high glucose when compared with those incubated with either the l-glucose or untreated controls. When HRP were incubated with an NO· donor, DETANONOATE ((Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate), we observed both decreased glyoxalase I expression and activity relative to untreated control cells. Further studies showed that HRP underwent apoptosis when incubated with DETANONOATE and that apoptosis increased further on co-incubation with high glucose. Our findings indicate that glyoxalase I is critical for pericyte survival under hyperglycemic conditions, and its inactivation and/or down-regulation by NO· may contribute to pericyte death by apoptosis during the early stages of diabetic retinopathy.