J Vet Sci.  2013 Sep;14(3):345-353. 10.4142/jvs.2013.14.3.345.

Black rice anthocyanidins prevent retinal photochemical damage via involvement of the AP-1/NF-kappaB/Caspase-1 pathway in Sprague-Dawley Rats

• Jia H
• Chen W
• Yu X
• Wu X
• Li S
• Liu H
• Liao J
• Liu W
• Mi M
• Liu L
• Cheng D

• KMID: 1705566
• DOI: http://doi.org/10.4142/jvs.2013.14.3.345

Abstract

The effects of black rice anthocyanidins (BRACs) on retinal damage induced by photochemical stress are not well known. In the present study, Sprague-Dawley rats were fed AIN-93M for 1 week, after which 80 rats were randomly divided into two groups and treated with (n = 40) or without BRACs (n = 40) for 15 days, respectively. After treatment, both groups were exposed to fluorescent light (3,000 +/- 200 lux; 25degrees C), and the protective effect of dietary BRACs were evaluated afterwards. Our results showed that dietary BRACs effectively prevented retinal photochemical damage and inhibited the retinal cells apoptosis induced by fluorescent light (p < 0.05). Moreover, dietary BRACs inhibited expression of AP-1 (c-fos/c-jun subunits), up-regulated NF-kappaB (p65) expression and phosphorylation of IkappaB-alpha, and decreased Caspase-1 expression (p < 0.05). These results suggest that BRACs improve retinal damage produced by photochemical stress in rats via AP-1/NF-kappaB/Caspase-1 apoptotic mechanisms.

Keyword

apoptosis; black rice anthocyanidins; Caspase-1; photochemical damage; retina

MeSH Terms

Animal Feed/analysis
Animals
Anthocyanins/administration & dosage/*pharmacology
Antioxidants/administration & dosage/*physiology
Blotting, Western
Caspase 1/*genetics/metabolism
Diet
Dietary Supplements/analysis
I-kappa B Proteins/genetics/metabolism
NF-kappa B/*genetics/metabolism
Neoplasm Proteins/genetics/metabolism
Nucleocytoplasmic Transport Proteins/genetics/metabolism
Oryza sativa/chemistry
Proto-Oncogene Proteins c-fos/genetics/metabolism
Proto-Oncogene Proteins c-jun/genetics/metabolism
Rats
Rats, Sprague-Dawley
Real-Time Polymerase Chain Reaction
Retinal Diseases/etiology/*prevention & control
Signal Transduction/*drug effects/radiation effects
Transcription Factor AP-1/*genetics/metabolism
Anthocyanins
Antioxidants
Caspase 1
I-kappa B Proteins
NF-kappa B
Neoplasm Proteins
Nucleocytoplasmic Transport Proteins
Proto-Oncogene Proteins c-fos
Proto-Oncogene Proteins c-jun
Transcription Factor AP-1

Figure

• 

  Fig. 1 Apoptotic index of the retina of rats treated with (-▪-) or without (-♦-) 1% BRACs for 15 days and exposed to light for 0~24 h. Values are the means for seven rats at each time point, with standard deviations represented by vertical bars. *Mean value was significantly different compared with the same light exposure time in rats not treated with BRACs (p < 0.05, t test). a,b,cMean values with different letters on the same curve at different exposure times were significantly different (p < 0.05, Ryan's multiple range test).

• 

  Fig. 2 The relative Caspase-1 (A) mRNA expression normalized for corresponding GAPDH levels in retinas of rats treated with (-▪-) or without (-♦-) 1% BRACs for 15 days and exposed to light for 0~24 h. The semi-quantitative densitometry of Caspase-1 (B) protein expressions normalized for 0 h light levels without BRACs (set as 1) in retinas of rats treated with (-▪-) or without (-□-) 1% BRACs for 15 days and exposed to light for 0 ~ 24 h. Values are the means for three determinations with their standard deviations shown by vertical bars for each time point. a,b,cMean values with unlike letters on the same curve or among the same diet group at different exposure times were significantly different by Ryan's multiple range test (p < 0.05). *Mean value was significantly different from that for rats at the same light exposure time but not treated with BRACs (p < 0.05, t test).

• 

  Fig. 3 The relative NF-κB, p65, (A) mRNA expressions normalized for corresponding GAPDH levels in retinas of rats treated with (-▪-) or without (-♦-) 1% BRACs for 15 days and exposed to light for 0 ~ 24 h. The semi-quantitative densitometry of Caspase-1 (B) protein expressions normalized for 0 h light levels without BRACs (set as 1) in retinas of rats treated with (-▪-) or without (-□-) 1% BRACs for 15 days and exposed to light for 0~24 h. Values are the means for three determinations with their standard deviations shown by vertical bars for each time point. a,b,cMean values with unlike letters on the same curve or among the same diet group at different exposure times were significantly different by Ryan's multiple range test (p < 0.05). *Mean value was significantly different from that for rats at the same light exposure time but not treated with BRACs (p < 0.05, t test).

• 

  Fig. 4 Phosphorylation of IκB-α protein exposed to light for 0 ~ 24 h. (A) after exposure to light, the phosphorylation of IκB-α was analyzed at various time points with western blot and (B) semi-quantitative densitometry of phosphor-IκB-α relative to 0 h light levels (set as 1) with (-▪-) or without (-□-) 1% BRACs for 15 days. a,b,cMean values with unlike letters among the same diet group at different exposure times were significantly different (p < 0.05, Ryan's multiple range test). *Mean value was significantly different compared with rats not treated with BRACs at the same light exposure time (p < 0.05, t test).

• 

  Fig. 5 The relative AP-1, c-fos, (A) mRNA expressions normalized for corresponding GAPDH levels in retinas of rats treated with (-▪-) or without (-♦-) 1% BRACs for 15 days and exposed to light for 0 ~ 24 h. The semi-quantitative densitometry of Caspase-1 (B) protein expressions normalized for 0 h light levels without BRACs (set as 1) in retinas of rats treated with (-▪-) or without (-□-) 1% BRACs for 15 days and exposed to light for 0 ~ 24 h. Values are the means for three determinations with their standard deviations shown by vertical bars for each time point. a,b,cMean values with unlike letters on the same curve or among the same diet group at different exposure times were significantly different by Ryan's multiple range test (p < 0.05). *Mean value was significantly different from that for rats at the same light exposure time but not treated with BRACs (p < 0.05, t test).

• 

  Fig. 6 The relative AP-1, c-jun, (A) mRNA expressions normalized for corresponding GAPDH levels in retinas of rats treated with (-▪-) or without (-♦-) 1% BRACs for 15 days and exposed to light for 0 ~ 24 h. The semi-quantitative densitometry of Caspase-1 (B) protein expressions normalized for 0 h light levels without BRACs (set as 1) in retinas of rats treated with (-▪-) or without (-□-) 1% BRACs for 15 days and exposed to light for 0~24 h. Values are the means for three determinations with their standard deviations shown by vertical bars for each time point. a,b,cMean values with unlike letters on the same curve or among the same diet group at different exposure times were significantly different by Ryan's multiple range test (p < 0.05).*Mean value was significantly different from that for rats at the same light exposure time treated but not treated with BRACs (p < 0.05, t test).

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