Immumohistochemistry staining for collagen type I.

<p>(A) Representative images. Brown region represents positive immunostaining for collagen type I which was substantially increased in diabetic rats and reduced by DiOHF treatment. The amount of collagen type I was found to be similar in control and control-treated rats. Collagen type I interstitial accumulation as assessed by percentage proportional area showing positive immunostaining in control and diabetic rats treated with or without DiOHF (B). Data expressed as mean ± SE. *<i>P</i>&lt;0.05 vs control (non-diabetic) rats; †<i>P</i>&lt;0.05 vs diabetic rats. Original magnification ×200.</p

Representative images for myocyte hypertrophy.

<p>(A) Diabetic rats demonstrated myocyte hypertrophy as evidenced by increased cross sectional area when compared with control rats. Treatment with DiOHF reduced cross sectional area in diabetic rats but had no effect on control rats. (B) Quantitative data for myocyte cross sectional area. *<i>P</i>&lt;0.05 vs control (non-diabetic) rats; †<i>P</i>&lt;0.05 vs diabetic rats. Original magnification ×200.</p

Txnip protein expression.

<p>(A) Representative western blot and (B) quantitation of Txnip normalized to loading control pan-actin. The significant increase in Txnip protein in diabetic rats was moderately reduced by DiOHF treatment. Data expressed as mean ± SE. *<i>P</i>&lt;0.05 vs control (non-diabetic) rats.</p

Representative pressure volume (PV) loops during preload reduction.

<p>Note the steeper slope of EDPVR (green line) and rightward shift in the diabetic group (C) compared to control (A). An increase in the slope of EDPVR indicated decreased chamber compliance as seen in the diabetic animals which was reduced with DiOHF treatment (D). *<i>P</i>&lt;0.05 vs control (non-diabetic) rats; †<i>P</i>&lt;0.05 vs diabetic rats.</p

Immunohistochemistry staining for collagen type III.

<p>(A) Representative images. Brown region represents positive immunostaining for collagen type III which was substantially increased in diabetic rats and reduced by DiOHF treatment. The amount of collagen type III was found to be similar in control and control-treated rats. Collagen type III interstitial accumulation as assessed by proportional area on sections showing positive immunostaining in control and diabetic rats treated with or without DiOHF (B). Data expressed as mean ± SE. Data expressed as mean ± SE. *<i>P</i>&lt;0.05 vs control (non-diabetic) rats; †<i>P</i>&lt;0.05 vs diabetic rats. Original magnification ×200.</p

Measurements of oxidative stress.

<p>Representative images for the localization of 3-nitrotyrosine in LV tissues, as assessed by the proportional area of positive immunostaining in control and diabetic rats treated with or without DiOHF (A). The marked increase in positive immunostaining (brown region) for 3-nitrotyrosine in diabetic rats was reduced by DiOHF treatment. The amount of 3-nitrotyrosine was found to be comparable in control and control-treated Ren-2 rats (B). Original magnification ×200. (C) NADPH-activated superoxide production in LV tissues as measured by lucigenin-enhanced chemiluminescence. Data expressed as mean ± SE. *<i>P</i>&lt;0.05 vs control (non-diabetic) rats; †<i>P</i>&lt;0.05 vs diabetic rats.</p

Intracellular antioxidants mRNA gene expression.

<p>(A) Measurement of Cu/Zn-superoxide dismutase (SOD1) and (B) gluthathione peroxidase (Gpx1) mRNA by real time RT-PCR, normalized to the housekeeping gene 18s. Diabetic rats demonstrated increased gene expression of SOD1 and Gpx1, which were not affected by DiOHF. Data expressed as mean ± SE. *<i>P</i>&lt;0.001 vs control (non-diabetic) rats.</p

Echocardiographic parameters in control and diabetic rats, treated with or without DiOHF for 6 weeks.

<p>Values are Mean ± SE; FAC, Fractional Area Change; LVID d, Left Ventricular Internal end Diastolic dimension; LVID s, Left Ventricular Internal end Systolic dimension; EF, Ejection Fraction; FS, Fractional Shortening; Dec T, Deceleration time.</p><p>*<i>P</i>&lt;0.05 vs control (non-diabetic) rats;</p>†<p><i>P</i>&lt;0.05 vs diabetic rats.</p

CXCR4 and SDF-1 mRNA in sham-operated and subtotally nephrectomized (SNx) rats.

<p>CXCR4 expression is increased in the kidneys of SNx rats (A), whereas SDF-1 is reduced (B). The kidneys of SNx rats also demonstrated an upregulation of the pro-fibrotic growth factor, transforming growth factor-β (TGF-β) (C). In cultured NRK-49F renal fibroblasts, recombinant TGF-β downregulated SDF-1 mRNA, with this effect being attenuated by HDAC inhibition with vorinostat (D). *p<0.001 vs. sham, ^†p<0.05 vs. sham, ^‡p<0.01 vs. sham, ^§p<0.001 vs. control, ^¶p<0.001 vs. TGF-β.</p

Functional characteristics of sham and subtotal nephrectomy (SNx) rats treated with vehicle or AMD3100.

<p>SBP = systolic blood pressure.</p><p>*p<0.05 vs. sham + vehicle, ^†p<0.05 vs. sham + AMD3100, ^‡p<0.01 vs. sham + vehicle, ^§p<0.01 vs. sham + AMD3100, ^¶p<0.001 vs. sham + vehicle, ^||p<0.05 vs. SNx + vehicle, **p<0.01 vs. SNx + vehicle.</p