1.Screening of differential proteins binding to Nox1 promoter in A549 cell model of inflammation and oxidative stress.
Xian QIU ; Shuiwang HU ; Jun XU ; Li LI ; Wenjie HUANG
Journal of Southern Medical University 2013;33(5):703-707
OBJECTIVETo screen the regulatory proteins involved in Nox1 promoter activation in a cell model of inflammation and oxidative stress.
METHODSA cell model of inflammation and oxidative stress was established by stimulating A549 cells with tumor necrosis factor-α (TNF-α). The differential proteins binding to Nox1 promoter were screened by DNA pull-down and the binding proteins were separated by 2D electrophoresis and selected according to the their differential expression levels (with over 1.5-fold changes relative to the control level). The screened proteins were finally identified by MALDI-TOF/TOF-MS.
RESULTSSeven differentially expressed protein spots (all upregulated in the cell model) were obtained, among which GLE1, DDX19A, KRT1 and KRT10 were identified by mass spectrometry.
CONCLUSIONGLE1, DDX19A, KRT1 and KRT10 participate in the activation of Nox1 promoter in TNF-α-induced A549 cells, and this result provides new insights into the biological roles of the regulatory proteins of Nox1 promoter in inflammation and oxidative stress.
Cell Line, Tumor ; DEAD-box RNA Helicases ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Inflammation ; Keratin-1 ; metabolism ; Keratin-10 ; metabolism ; Mass Spectrometry ; NADPH Oxidase 1 ; NADPH Oxidases ; genetics ; metabolism ; Nucleocytoplasmic Transport Proteins ; metabolism ; Oxidative Stress ; Promoter Regions, Genetic ; Tumor Necrosis Factor-alpha ; adverse effects
2.Black rice anthocyanidins prevent retinal photochemical damage via involvement of the AP-1/NF-kappaB/Caspase-1 pathway in Sprague-Dawley Rats.
Hao JIA ; Wei CHEN ; Xiaoping YU ; Xiuhua WU ; Shuai LI ; Hong LIU ; Jiru LIAO ; Weihua LIU ; Mantian MI ; Longjian LIU ; Daomei CHENG
Journal of Veterinary Science 2013;14(3):345-353
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.
Animal Feed/analysis
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Animals
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Anthocyanins/administration & dosage/*pharmacology
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Antioxidants/administration & dosage/*physiology
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Blotting, Western
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Caspase 1/*genetics/metabolism
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Diet
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Dietary Supplements/analysis
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I-kappa B Proteins/genetics/metabolism
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NF-kappa B/*genetics/metabolism
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Neoplasm Proteins/genetics/metabolism
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Nucleocytoplasmic Transport Proteins/genetics/metabolism
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Oryza sativa/chemistry
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Proto-Oncogene Proteins c-fos/genetics/metabolism
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Proto-Oncogene Proteins c-jun/genetics/metabolism
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Rats
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Rats, Sprague-Dawley
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Real-Time Polymerase Chain Reaction
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Retinal Diseases/etiology/*prevention & control
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Signal Transduction/*drug effects/radiation effects
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Transcription Factor AP-1/*genetics/metabolism