Vitamin e treatment prevents diabetes-induced abnormality in retinal blood flow possibly through the diacylglycerol-protein kinase c pathway. Kunisaki, Makoto, Sven-Erik Bursell, Allen C. Clermont, Hidehiro Ishii, Lawrence M. Ballas, Michael R. Jirousek, Fumio Umeda, Hajime Nawata, and George L. King. Research Division, Joslin Diabetes Center, and Departments of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02215, Third Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka 812, Japan, Sphinx Pharmaceuticals Corporation, Durham, NC 27717, and Lilly Research Laboratories, Inc., Indianapolis, IN 46285
APStracts 2:0040E, 1995.
We have characterized the effects of d-[alpha]-tocopherol (vitamin E) on the activation of protein kinase C (PKC) and diacylglycerol (DAG) levels in retinal tissues of diabetic rats, and correlated its effects to diabetes-induced changes in retinal hemodynamics. Comparing streptozotocin-induced diabetic rats to controls, membrane PKC specific activities were increased by 71% (P<0.05). Western blot analysis showed membrane PKC [beta]II isoform was increased by 133+45% (P<0.05). Intraperitoneal injection of d-[alpha]-tocopherol (40 mg/kg) every other day prevented the increases in membrane PKC specific activity and PKC [beta]II protein by immunoblots. Diabetes -induced increases in DAG levels were also normalized by d-[alpha] -tocopherol treatment of 2 weeks duration. Physiologically, angiographic abnormalities of retinal hemodynamics based on computerized video based fluorescein angiography and assoicated with increases of DAG and membranous PKC levels were also prevented by d -[alpha]-tocopherol treatment in diabetic rats. The effect of d -[alpha]-tocopherol on retinal vascular cells was also studied. Exposure of retinal endothelial cells (REC) to 22 mM glucose for three days compared to 5 mM glucose, increased total DAG and [3H] -palmitate labeled DAG levels by 35%+8% and 50+8% (P<0.05), respectively, more than REC exposed to 5.5 mM glucose. The presence of d-[alpha]-tocopherol (50[mu]g/ml) prevented the increases in both total DAG and [3H]-palmitate labeled DAG levels in cells exposed to 22 mM glucose. These findings suggested that treatment with d -[alpha]-tocopherol can prevent diabetes-induced abnormalities in rat retinal blood flow. The mechanism of the d-[alpha]-tocopherol's effect appears to be mediated by the normalization of hyperglycemia -induced activation of DAG-PKC pathway in the retina of diabetic rats.

Received 18 August 1994; accepted in final form 27 February 1995.
APS Manuscript Number E334-4.
Article publication pending Am. J. Physiol. (Endocrinol. Metab.).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 10 March 1995.