Fenofibrate prevents high glucose-induced oxidative stress and apoptosis in HSCs under VEGF inhibition.

<p>Marked increases in TUNEL-positive human Schwann cells (HSCs) were observed in the 30 Glu group and 30 Glu VEGFR12 compared to the 5 Glu group with or without fenofibrate treatment. Quantitative analyses of the results are shown (A and B, original magnification, ×1,000). **<i>P</i><0.01 and ^#<i>P</i><0.001 versus 5 Glu, 5 Glu VEGFR12, or 5 Glu VEGFR12+Feno groups. Western blot analysis of Bcl-2, Bax, and β-actin and quantitative analyses of the results are shown (C and D). *<i>P</i><0.05 and **<i>P</i><0.01 versus <i>db/m</i> controls. Analysis of intracellular PPARα-phospho-The¹⁷²/total AMPK-PGC-1α pathway in cultured HSCs. Representative Western blot of PPARα, phospho-The¹⁷²/total AMPK, PGC-1α, and β-actin and quantitative analyses of the results are shown (E to H). *<i>P</i><0.05 and **<i>P</i><0.01 versus other groups. 5 Glu; 5 mM D-glucose concentration, 5 Glu VEGFR12; 5 mM D-glucose concentration with VEGFR12 inhibition, 5 Glu VEGFR12+Feno; 5 mM D-glucose concentration with VEGFR12 inhibition+Feno; IV, 30 Glu; 30 mM glucose, 30 Glu VEGFR12; 30 mM D-glucose concentration with VEGFR12 inhibition; 30 Glu VEGFR12+Feno; 30 mM D-glucose concentration with VEGFR12+Feno.</p

Body weight and biochemical characteristics of <i>db/m</i> and <i>db/db</i> mice under VEGFR inhibition treated with fenofibrate.

<p>Abbreviations: Feno; fenofibrate, TC; total cholesterol, HDL-C; high density lipoprotein-cholesterol, TG; triacylglycerol, VEGFR12; VEGFR-1 and –2 inhibition.</p><p><i>P</i><0.01,</p><p><i>P</i><0.001 compared with <i>db/m</i>, <i>db/m</i>+VEGFR12, and <i>db/m</i>+VEGFR12+Feno groups.</p><p><i>P</i><0.05 compared with <i>db/db</i> and <i>db/db</i>+VEGFR12 groups.</p

Fenofibrate attenuates fibrosis and ischemia of the sciatic nerve in <i>db/db</i> mice with VEGF inhibition.

<p>Nerve fibrosis, pro-fibrotic transforming growth factor (TGF)-β and platelet endothelial cell adhesion molecule (PECAM)-1 expression in the sciatic nerves of <i>db/m</i> and <i>db/db</i> mice after 12 weeks of VEGFR inhibition plus fenofibrate or VEGFR inhibition alone. Representative Masson's trichrome staining and quantitative analysis of the fibrotic area (1000×) (A). Representative immunohistochemical staining for TGF-β1 and PECAM-1 (mouse endothelial cell maker) and quantitative analyses of the results are shown (C to F). Representative Western blot analysis of TGF-β1, PECAM-1 and hypoxia-inducible factor (HIF)-1α (a marker for hypoxia) and quantitative analyses of the results are shown (G to J). *<i>P</i><0.05 and **<i>P</i><0.01 versus <i>db/m</i> controls, <i>db/m</i>+Feno, <i>db/m</i> VEGFR12, <i>db/m</i> VEGFR12+Feno, <i>db/db</i>+Feno and <i>db/db</i> VEGFR12+Feno mice (A to D). *<i>P</i><0.05, **<i>P</i><0.01 and ^#<i>P</i><0.001 versus <i>db/m</i> controls.</p

Fenofibrate upregulates the expression of PPARα-PGC-1α-phospho-The¹⁷² AMPK in <i>db/db</i> mice with VEGF inhibition.

<p>Peroxisome proliferative-activated receptor (PPAR)α, PPARγ coactivator (PGC)-1α, phospho-The¹⁷²/total AMPK and β-actin expression in the sciatic nerves of <i>db/m</i> and <i>db/db</i> mice after 12 weeks VEGFR inhibition plus fenofibrate or VEGFR inhibition alone. Representative immunohistochemical staining of PPARα and quantitative analyses of the results are shown (A and B). *<i>P</i><0.05 versus <i>db/m</i> controls. Western blot analysis of PPARα, PGC-1α, phospho-The¹⁷²/total AMPK and β-actin (C, respectively) and quantitative analyses of the results are shown. *<i>P</i><0.05 and **<i>P</i><0.01 versus <i>db/m</i> controls.</p

Fenofibrate improves sensory and motor functions of the sciatic nerve in <i>db/db</i> mice with VEGF inhibition.

<p>Tactile threshold (A) and motor conduction latency of the sciatic nerves (B) in <i>db/m</i> and <i>db/db</i> mice after 12 weeks of vascular endothelial growth factor receptor (VEGFR) inhibition plus fenofibrate or VEGFR inhibition alone. *<i>P</i><0.05, **<i>P</i><0.01 and ^#<i>P</i><0.001 versus <i>db/m</i> controls, <i>db/m</i>+Feno, <i>db/m</i> VEGFR12, <i>db/m</i> VEGFR12+Feno, <i>db/db</i>+Feno, and <i>db/db</i> VEGFR12+Feno mice.</p

Fenofibrate decreases inflammatory cell infiltration, oxidative stress and apoptosis of the sciatic nerve in <i>db/db</i> mice with VEGF inhibition.

<p>Representative immunohistochemical stain for F4/80-, 8-OH-dG- and TUNEL-positive cells and quantitative analyses of the results are shown (A to F). *<i>P</i><0.05, **<i>P</i><0.01 and ^#<i>P</i><0.001 versus <i>db/m</i> controls. Representative electron microscopic images of the sciatic nerve endoneural endothelial cells (15,000×) and nerve bundles (5,000×) (G and H). Disorganized, vacuolized and thickened endothelial cells were observed in <i>db/db</i> controls and <i>db/db</i> VEGFR12 mice. These characteristics decreased after 12-week fenofibrate treatment in the <i>db/db</i>+Feno and <i>db/db</i> VEGR12+Feno mice. A marked decrease in the number of unmyelinated nerve bundles (red arrowheads) and prominent axonal degeneration (red arrows) were observed in the <i>db/db</i> controls and <i>db/db</i> VEGFR12 mice. These deficits were also improved by the 12-week fenofibrate treatment in the <i>db/db</i>+Feno and <i>db/db</i> VEGR12+Feno mice. Quantitative analyses of number of endoneurial blood vessel, area of unmyelinated fiber, axonal diameter and number of degenerated axon are shown (I to L). *<i>P</i><0.05 versus other groups, *<i>P</i><0.05, **<i>P</i><0.01, and ^#<i>P</i><0.001 versus <i>db/m</i> controls. Representative immunofluorescent stains for PECAM-1 and TUNEL-double positive cells in <i>db/db</i> mice groups and quantitative analyses of the results are shown (M and N). An increase in the number of PECAM-1 and TUNEL-double positive cells (white arrows) was observed in the <i>db/db</i> controls and <i>db/db</i> VEGFR12 mice. **<i>P</i><0.01 and ^#<i>P</i><0.001 versus <i>db/m</i> controls.</p

Fenofibrate prevents high glucose-induced oxidative stress and apoptosis in HUVECs under VEGF inhibition.

<p>Marked increases in TUNEL-positive human umbilical vein endothelial cells (HUVECs) were observed in the 30 Glu group and 30 Glu VEGFR12 compared to the 5 Glu group with or without fenofibrate treatment. Quantitative analyses of the results are shown (A and B). *<i>P</i><0.05, **<i>P</i><0.01 and ^#<i>P</i><0.001 versus 5 Glu, 5 Glu VEGFR12, or 5 Glu VEGFR12+Feno groups. Analysis of intracellular PPARα-PGC-1α-phospho-The¹⁷²/total AMPK pathway and PI3K-Akt-eNOS signaling and apoptosis in cultured HUVECs. Representative Western blot of PPARα, PGC-1α and phospho-The¹⁷²/total AMPK, and β-actin and quantitative analyses of the results are shown (C to F). Representative Western blot of PI3K, total and phospho-Ser⁴⁷³ Akt, total and phospho-Ser¹¹⁷³ eNOS, and β-actin and quantitative analyses of the results are shown (G to J). The NOx concentrations from the supernatant of HUVECs are shown (K). *<i>P</i><0.05 and **<i>P</i><0.01 versus other groups. 5 Glu; 5 mmol/L D-glucose concentration, 5 Glu VEGFR12; 5 mmol/L D-glucose concentration with VEGFR12 inhibition, 5 Glu VEGFR12+Feno; 5 mmol/L D-glucose concentration with VEGFR12 inhibition+Feno; IV, 30 Glu; 30 mmol/L glucose, 30 Glu VEGFR12; 30 mmol/L D-glucose concentration with VEGFR12 inhibition; 30 Glu VEGFR12+Feno; 30 mmol/L D-glucose concentration with VEGFR12+Feno.</p