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KO islets accumulate oxidative stress, inflammation, and fibrosis.

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posted on 2015-10-15, 03:22 authored by Justin R. Hassler, Donalyn L. Scheuner, Shiyu Wang, Jaeseok Han, Vamsi K. Kodali, Philip Li, Julie Nguyen, Jenny S. George, Cory Davis, Shengyang P. Wu, Yongsheng Bai, Maureen Sartor, James Cavalcoli, Harmeet Malhi, Gregory Baudouin, Yaoyang Zhang, John R. Yates III, Pamela Itkin-Ansari, Niels Volkmann, Randal J. Kaufman

(A) mRNA-Seq expression values for 25/368 of the mRNAs identified by Venn analysis (Fig 3C; right panel, underlined) that are reduced by Ire1α because of glucose that accumulates in the KOFe/-; Cre ([n = 5], [p-values ≤ 0.05]). (B) Oxidized lipid (hydroxyl-octadecadienoic acids, HODEs) from islets of the indicated genotypes infected with Ad-Cre Ad-GFP or no virus control ([n = 2; controls versus n = 3; Ad-Cre], [p = 0.00434]). (C) Antinitrotyrosine immunohistochemistry (IHC) of islets from 8-mo-old WTFe/Fe and KOFe/Fe; Cre mice 15 wk post-Tam with or without BHA diet for 3 wk. (Scale bar, 50 μm) (WTFe/Fe [n = 4 with BHA], [n = 5 regular chow]), (KOFe/Fe; Cre [n = 5 with BHA], [n = 6 regular chow]). (p = 0.00698; WTFe/Fe versus WTFe/Fe with BHA), (p = 0.04018; WTFe/Fe versus KOFe/Fe; Cre) and (p = 0.04420; KOFe/Fe; Cre versus KOFe/Fe; Cre with BHA). (D) Masson’s trichrome stain (blue) for collagens. Results demonstrate increased staining surrounding KOFe/Fe; Cre islets with haemotoxylin (red) and eosin (black) co-stains. Quantification of percent strong collagen stain is shown below the images. Scale bar, 50 μm. (WTFe/Fe [n = 4 with BHA], [n = 5 regular chow]), (KOFe/Fe; Cre [n = 5 with BHA], [n = 6 without BHA]). Percent strong collagen stain significance for WTFe/Fe without BHA versus KOFe/Fe; Cre without BHA p = 0.01049). (E) 8-mo-old male mice carrying the doubly floxed allele (Ire1αFe/Fe) with and without RIP-Cre 12 wk post-Tam had their pre-BHA GTTs taken, and then half were fed the antioxidant BHA supplemented chow diet for 3 wk or not before examining the mice by GTT again. (WTFe/Fe [n = 11 with BHA], [n = 12 regular chow], [p = 0.035]), (KOFe/Fe; Cre [n = 18 with BHA], [n = 16 without BHA], [p = 0.041]). P-values were calculated by one-tailed student’s t test comparison of the areas under the GTT curves for the biological replicates of control group WTFe/Fe versus the Tam-induced KOFe/Fe; Cre group.