Figure 3.

GY supplementation and oxidative stress and MMP-9 activation. Mice retinal microvessels prepared by osmotic shock were analyzed for oxidative stress by quantifying (a) ROS levels using DCFDA and (b) protein carbonyls employing 2,4-dinitrophenylhydrazine. (c) MMP-9 activity was measured using its monoclonal antibody and a fluorogenic substrate. (d) Representative image of the co-localization of homocysteine and MMP-9 using DyLight-488 (green) and Texas Red (red) conjugated secondary antibodies for homocysteine and MMP-9, respectively. Each measurement was made in duplicate in 6 to 7 mice/group, and the values are represented as mean ± SD. Norm, normal; Diab and Diab/GY, diabetic mice without or with GY respectively; Diab/H and Diab/H + GY, homocysteine supplemented diabetic mice, without or with GY. *P < 0.05 versus normal, #P < 0.05 versus diabetes, and ^P < 0.05 versus diabetic mice receiving homocysteine.