Efficacy of troxerutin on streptozotocin-induced rat model in the early stage of diabetic retinopathy.
The vascular changes associated with early diabetic retinopathy, which include the formation of microaneurysms and acellular capillaries, vessel dilation, vascular endothelial growth factor expression, were investigated experimentally in streptozotocin-induced diabetic rats treated with antioxidants: troxerutin (trihydroxy-ethylrutoside, CAS 7085-55-4), Vaccinium myrtillus, and calcium dobesilate (hydroquinone calcium sulfonate, CAS 20123-80-2). The development and progression of retinopathy was followed using fundus photography. After 3 months, the rats were sacrificed and half of the eyes were prepared for neovascularization analysis, and the other half were used for VEGF (vascular endothelial growth factor) analysis. The results from fundus photography and ADPase (adenosine diphosphatase) staining were quantified by the percentage area of the retinal vasculature using a commercial image analyzer. The VEGF protein in the retinal homogenates was assessed using an ELISA (enzyme linked immunosorbent assay) kit and VEGF-mRNA by RT-PCR (reverse transcription polymerase chain reaction). In the ADPase stain, the retinal vascular percent area increased significantly in the diabetic control. Neovascularization and aneurysms were observed in the diabetic control and were attenuated by 50 mg/kg troxerutin, but the retinal vascular percentage area was not significantly different from the diabetic control. The VEGF protein concentration was higher in diabetic rats than in the nondiabetic rats (21.5 +/- 2.1 vs 27.7 +/- 5.8 pg/mg, p < 0.05), and this increase was attenuated by 10 mg/kg troxerutin (24.5 +/- 3.8 pg/mg, p < 0.05) and prevented by 50 mg/kg troxerutin (19.5 +/- 2.2 pg/mg, p < 0.05). However, there were no significant differences between the groups. The VEGF-mRNA density showed a increasing tendency by 20% in the diabetic rats compared with the non-diabetic rats (1.0 +/- 0.1 vs 1.2 +/- 0.1 VEGF/beta-actin), and this increase was corrected by 10 mg/kg troxerutin (1.0 +/- 0.1 VEGF/beta-actin), 50 mg/kg troxerutin (0.9 +/- 0.1 VEGF/beta-actin) and Vaccinium myrtillus (1.1 +/- 0.1 VEGF/beta-actin). Oxidative stress might be involved in the upregulation of retinal VEGF during early diabetes, and it is likely that troxerutin has comparatively effective antioxidant properties. Therefore, troxerutin might be a useful treatment for attenuating diabetic retinopathy.
Chung
Han Kook
HK
Research Institutes of Dong-A Pharmaceutical Company, Kyunggi, Republic of Korea. ahnbo@donga.co.kr
Choi
Seul Min
SM
Ahn
Byoung Ok
BO
Kwak
Hyun Hee
HH
Kim
Jeong Hoon
JH
Kim
Won Bae
WB
D000818:Animals; D001081:Apyrase / Q000378:metabolism; D001786:Blood Glucose / Q000378:metabolism; D001835:Body Weight / Q000187:drug effects; D002123:Calcium Dobesilate / Q000627:therapeutic use; D003921:Diabetes Mellitus, Experimental / Q000473:pathology*; D003930:Diabetic Retinopathy / Q000188:drug therapy* / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005654:Fundus Oculi; D006490:Hemostatics / Q000627:therapeutic use; D006896:Hydroxyethylrutoside / Q000031:analogs & derivatives* / Q000627:therapeutic use; D008517:Phytotherapy; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D029800:Vaccinium myrtillus; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D014662:Vasoconstrictor Agents / Q000627:therapeutic use*
D001786:Blood Glucose; D006490:Hemostatics; D006896:Hydroxyethylrutoside; D042461:Vascular Endothelial Growth Factor A; D014662:Vasoconstrictor Agents; D002123:Calcium Dobesilate; D012313:RNA; C005865:troxerutin; D001081:Apyrase