JCI - Arsenic-stimulated liver sinusoidal capillarization in mice requires NADPH oxidase

Arsenic-stimulated liver sinusoidal capillarization in mice requires NADPH oxidase–generated superoxide

Adam C. Straub, Katherine A. Clark, Mark A. Ross, Ashwin G. Chandra, Song Li, Xiang Gao, Patrick J. Pagano, Donna B. Stolz, Aaron Barchowsky

Published in Volume 118, Issue 12

J Clin Invest. 2008; 118(12):3980–3989 doi:10.1172/JCI35092

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Figure 3

Arsenic inhibits SEC scavenging of acylated protein.

(A) Mice were injected in the tail vein with 150 mg/kg FITC-labeled succinylated BSA (suc-BSA) or BSA (both green) in 200 μl saline. After 10 min, the mice were euthanized, and livers were excised, fixed in 4% paraformaldehyde, and sectioned for confocal microscopic analysis. Sections were stained with rhodamine-conjugated antibody to PECAM-1 (red) and DRAQ5 (blue). Images are representative of sections from 3 mice in each group. LV, large vessel; S, sinusoid vessel. Scale bar: 10 μm. (B) FITC-labeled acetylated LDL and biotin-labeled succinylated BSA (3 ml; 150 mg/ml saline) were infused over 3 min into the vena cavas of untreated mice and mice exposed to 100 ppb arsenic (As) for 2 wk. Livers were then excised, frozen in liquid N₂, and sectioned, and total protein was extracted for assay of retained biotin label by immunoblotting, as described in Methods. Data are mean ± SD band density of biotin-labeled succinylated BSA normalized to β-actin (n = 3). NT, not treated. (C) Isolated SECs, incubated in the absence or presence of 1 mM tempol for 10 min, were left untreated or exposed to 2.5 μM arsenite for 24 h. Biotin-labeled succinylated BSA (20 μg/ml) was then added for 10 min, and after rinsing, total proteins were extracted for Western analysis (n = 3). Data are presented as in B. *P < 0.05, **P < 0.01 versus control; ^#P < 0.05 versus arsenic.