J Korean Med Sci.  2013 Apr;28(4):608-613. 10.3346/jkms.2013.28.4.608.

Chlorogenic Acid Decreases Retinal Vascular Hyperpermeability in Diabetic Rat Model

• Shin JY
• Sohn J
• Park KH

• KMID: 1786971
• DOI: http://doi.org/10.3346/jkms.2013.28.4.608

Abstract

To evaluate the effect of chlorogenic acid (CGA), a polyphenol abundant in coffee, on retinal vascular leakage in the rat model of diabetic retinopathy, Sprague-Dawley rats were divided into four groups: controls, streptozotocin-induced diabetic rats, and diabetic rats treated with 10 and 20 mg/kg chlorogenic acid intraperitoneally daily for 14 days, respectively. Blood-retinal barrier (BRB) breakdown was evaluated using FITC-dextran. Vascular endothelial growth factor (VEGF) distribution and expression level was evaluated with immunohistochemistry and Western blot analysis. Expression of tight junction proteins, occludin and claudin-5, and zonula occludens protein, ZO-1 was also evaluated with immunohistochemistry and Western blot analysis. BRB breakdown and increased vascular leakage was found in diabetic rats, with increased VEGF expression and down-regulation of occludin, claudin-5, and ZO-1. CGA treatment effectively preserved the expression of occludin, and decreased VEGF levels, leading to less BRB breakdown and less vascular leakage. CGA may have a preventive role in BRB breakdown in diabetic retinopathy by preserving tight junction protein levels and low VEGF levels.

Keyword

Chlorogenic Acid; Diabetic Retinopathy; Diabetes Rat Model; Vascular Endothelial Growth Factor; Blood-Retinal Barrier

MeSH Terms

Animals
Blood-Retinal Barrier/*drug effects
Chlorogenic Acid/metabolism/*pharmacology
Claudin-5/metabolism
Dextrans/chemistry
Diabetes Mellitus, Experimental/complications/metabolism/*pathology
Diabetic Retinopathy/etiology/prevention & control
Down-Regulation
Fluorescein-5-isothiocyanate/chemistry
Male
Occludin/metabolism
Rats
Rats, Sprague-Dawley
Retina/*metabolism
Tight Junction Proteins/metabolism
Vascular Endothelial Growth Factor A/metabolism
Zonula Occludens-1 Protein/metabolism
Claudin-5
Occludin
Tight Junction Proteins
Vascular Endothelial Growth Factor A
Zonula Occludens-1 Protein
Chlorogenic Acid
Fluorescein-5-isothiocyanate
Dextrans

Figure

• 

  Fig. 1 Leakage of retinal vessels with FITC-dextran injection. Chlorogenic acid decreased the hyperpermeability of retinal vessels in diabetic rats (scale bar: 100 µm). STZ, streptozotocin; CGA10, chlorogenic acid 10 mg/kg; CGA20, chlorogenic acid 20 mg/kg.

• 

  Fig. 2 VEGF expression with immunohistochemistry and Western blot analysis. VEGF expression in endothelial cells (arrows) was suppressed by chlorogenic acid administration in diabetic rats (scale bar: 50 µm). *P < 0.05 compared with control, †P < 0.05 compared with STZ. STZ, streptozotocin; CGA10, chlorogenic acid 10 mg/kg; CGA20, chlorogenic acid 20 mg/kg; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer.

• 

  Fig. 3 Expression of occludin, claudin-5, and ZO-1 with immunohistochemistry. Occludin, claudin-5, and ZO-1 staining in the vascular endothelium of the ganglion cell layer (arrows) were found to be decreased in the diabetic retina compared to control. With CGA 20 mg/kg administration, the decrease was restored, and staining of occludin and claudin-5 in the endothelial cell junctions were increased. However, ZO-1 expression remained grossly unchanged. (scale bar: 50 µm). STZ, streptozotocin; CGA10, chlorogenic acid 10 mg/kg; CGA 20, chlorogenic acid 20 mg/kg; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer.

• 

  Fig. 4 Expression of occludin, claudin-5, and ZO-1 with Western blot analysis. Occludin expression was decreased in diabetic rats, and restored with chlorogenic acid administration. Claudin-5 was decreased in diabetic rats, but did not increase to significant levels with chlorogenic acid administration. ZO-1 decreased in diabetic rats, but showed no change with chlorogenic acid administration. *P < 0.05 compared with control, †P < 0.05 compared with STZ. STZ, streptozotocin; CGA10, chlorogenic acid 10 mg/kg; CGA20, chlorogenic acid 20 mg/kg.

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