Fig 1.
Changes in body weight and levels of blood glucose from the STZ injection to the end of the 5th weekend.
(A) body weight; (B) blood glucose (*P < 0.05, versus Normal; #P < 0.05 versus STZ+ATWLPPR).
Fig 2.
ERG responses in the Normal, D+saline and D+ATWLPPR groups.
(A) Representative original recordings from the three groups; (B) Quantitative analysis of b-wave amplitudes (mV); (C) Quantitative analysis of b-wave implicit time (ms). *P < 0.05 versus the Normal group; #P < 0.05 versus the D+saline group. D, Diabetic.
Fig 3.
(A) Normal; (B) Diabetic; (C) Diabetic treated with ATWLPPR. Arrows: fluorescence leakage areas. Bar = 300 μm.
Fig 4.
Treatment with ATWLPPR reduced the leukocyte attachment.
Con A perfusion retinas show attached leukocytes (arrows) within the vasculature of diabetic retinas. Representative retina flat-mount images from (A) Normal, (B) D+saline and (C) D+ATWLPPR groups. (D) Quantitative analysis of leukocyte adhesion. Scale bar = 40 mm. *P < 0.05, versus the Normal group; #P < 0.05, versus the D+saline group. D, Diabetic.
Fig 5.
Diabetes-induced decrease in occludin was inhibited by treatment with ATWLPPR.
*P < 0.05, versus the Normal group; #P < 0.05, versus the D+saline group. D, Diabetic.
Fig 6.
Extravascular albumin was inhibited by ATWLPPR in diabetic retinas.
(A) Immunohistochemical staining shows the extravascular albumin (arrows) in retinal sections. Normal (a, d, g); D+saline (b, e, h); D+ATWLPPR (c, f, i). Scale bar = 50 mm. (B) Western blot analysis indicates the significant increase in leaking albumin in the D+saline retinas was suppressed by treatment with ATWLPPR. D, Diabetic. *P < 0.05, versus the Normal group; #P < 0.05, versus the D+saline group.
Fig 7.
The increase in ROS formation in diabetic retinas was inhibited by treatment with ATELPPR or pretreatment with apocynin (1 mM).
(A) Representative images of DHE imaging. (B) Quantitative analysis of fluorescence intensity in DHE images showed an obvious increase in ROS formation in diabetic retinas. ROS generation was significantly reduced in D+ATWLPPR and pretreatment with apocynin (D+apocynin). D, Diabetic. *P < 0.05, versus the Normal group; #P < 0.05, versus the D+saline group. Scale bar = 50 mm, 200X. GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer.
Fig 8.
ATWLPPR decreased oxidative stress in the diabetic retinas by regulation of the MDA levels and SOD activity.
(A) MDA levels (nmol MDA/mg protein); (B) SOD activity (U/mg protein). D, Diabetic. *P < 0.05, versus the Normal group; #P < 0.05, versus the D+saline group.
Fig 9.
ATWLPPR inhibited the diabetes-induced up-regulation of inflammatory proteins in the retina.
(A) Representative immunohistochemical images for ICAM-1 (a, b, c), GFAP (d, e, f), and VEGF (g, h, i) staining on retinal sections. Arrows, immune-positive. Scale Bar = 50 mm, 200X. GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer. (B) Western blot analysis of the GFAP, VEGF and ICAM-1 expression levels in retinas. D, Diabetic. *P < 0.05, versus the Normal group; #P < 0.05, versus the D+saline group.