Purchase this article with an account.
T. Hikichi, M. Sasaki, A. Yoshida, N. Shishido, M. Nakamura, K. Ishikawa, J. Iwamoto, Y. Hayashi; Nitric Oxide Mediates Glucose-Induced Apoptosis in Long-Term Cultured Retinal Pericytes . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3885.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: In the early diabetic retinopathy, the partial loss of pericytes have been often observed, whereas these cells in vitro are rather tough to high glucose stress. To address this discrepancy, we have extensively examined culture conditions, which would make pericytes more sensitive to high glucose. Methods: Retinal pericytes were prepared from bovine retinal capillaries and cultured under a high consentration of glucose. The nuclei were stained with the vital nuclear dye bisbenzamide (Hoechst 33342) and the percentage of apoptotic nuclei in total cells was counted. Results: We found that with increasing of both culture time and passage number, they become more susceptible to high glucose stress. Pericytes, which underwent through 1 to 5 passages and cultured for less than 12 weeks, showed neither any cellular degeneration, nor apoptosis when exposed to a high glucose condition for 8 days. In contrast, cells showed distinct apoptosis in response to a high concentration of glucose, when underwent more than 9 passages. The sister cultured cells did not show any damage at all in both normal and high concentrations of glucose and mannitol, respectively. In prior to this degenerative process, there were increased activities of antioxidant enzymes such as glutathione reductase, glutathione peroxidase and superoxide dismutase. Furthermore, the amount of NO2 in extracellular space was increased only in a high glucose condition, whereas the cytoplasmic pattern of NOS localization were altered in the presence of high concentrations of both glucose and mannitol. Treatment with L-NAME inhibited this high glucose induced apoptosis of cells with 9 passages. Conclusions: These data, taken together, suggest that pericytes with high number of passages have some pathological change common to that in the early diabetic retinopathy, where high glucose stress mediates apoptosis through increment in NO production.
This PDF is available to Subscribers Only