Trimetazidine reduces early and long-term effects of experimental renal warm ischemia: A dose effect study Jerome Cau, MD, Frederic Favreau, PhD, Jean.

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Trimetazidine reduces early and long-term effects of experimental renal warm ischemia: A dose effect study Jerome Cau, MD, Frederic Favreau, PhD, Jean Paul Tillement, MD, PhD, Lilach O. Lerman, MD, PhD, Thierry Hauet, MD, PhD, Jean Michel Goujon, MD, PhD Journal of Vascular Surgery Volume 47, Issue 4, Pages 852-860.e4 (April 2008) DOI: 10.1016/j.jvs.2007.10.036 Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1 Influence of trimetazidine on warm ischemia time on the glomerular filtration rate (A), urinary protein excretion (B). The values reported are the mean ±SE. *P < .05, **P < .01, ***P < .001 WI-60 and WI-60-TMZ1 mg groups vs WI-60-TMZ5 mg and WI-60-TMZ10 mg groups and WI-60-TMZ5 mg and WI-60-TMZ10 mg groups vs control and unif groups. Journal of Vascular Surgery 2008 47, 852-860.e4DOI: (10.1016/j.jvs.2007.10.036) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1, online only TMAO excretion in urine (C) and plasma levels (D). Renal function was measured in the different experimental groups after different warm ischemia and compared with control group and uninephrectomized group (unif). TMAO excretion was influenced by TMZ in urine (C) and plasma (D). The values reported are the mean ±SE. *P < 0.05, **P < 0.01, ***P < .001 WI-60 and WI-60-TMZ1 mg groups vs WI-60-TMZ5 mg and WI-60-TMZ10 mg groups and WI-60-TMZ5 mg and WI-60-TMZ10 mg groups vs control and unif groups. Journal of Vascular Surgery 2008 47, 852-860.e4DOI: (10.1016/j.jvs.2007.10.036) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 2 Western blot analysis of HIF-1α. HIF-1α expression was examined by western blot in cortex and outer medulla at days 0, 1, and 7, and 3 months after reperfusion. A, representative immunoblots for HIF-1α after experimental ischemia (see Material and Methods section) followed by reperfusion. HIF-1α expression in normal kidney (control) and uninephrectomized animals (unif) are compared under experimental conditions. B, Expression of HIF-1α determined at days 0 (D0), 1 (D1), and 7 (D7), and 3 months (M3) after reperfusion. Data were presented as means +SD from three independent experiments. *P < .05 TMZ-treated (5 and 10 mg/Kg) groups vs TMZ-untreated group, TMZ-treated (1 mg/Kg), **P < .01 TMZ-treated (5 and 10 mg/Kg) groups vs non-TMZ-treated group, TMZ-treated (1mg/Kg). Journal of Vascular Surgery 2008 47, 852-860.e4DOI: (10.1016/j.jvs.2007.10.036) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 3 Renal protein densitometry and representative immunoblots of the regulator of apoptosis at day 1 (top), day 7 (middle), and month 3 (bottom). *P < .05 vs normal and WI-60 or WI-60-TMZ1 mg. Journal of Vascular Surgery 2008 47, 852-860.e4DOI: (10.1016/j.jvs.2007.10.036) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 4, online only Effect of warm ischemia time inflammation in post reperfused kidney. A, CD4+ cells, and B, monocyte/macrophages. The number of positively stained per surface area were counted on biopsy samples from experimental groups. The values reported are the mean ± SE from 8 to 12 separate determinations. The values reported are the mean ±SE. *P < .05, **P < .01 WI-60 and WI-60-TMZ1 mg, vs WI-60-TMZ5 mg and WI-60-TMZ10 mg. Journal of Vascular Surgery 2008 47, 852-860.e4DOI: (10.1016/j.jvs.2007.10.036) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 5 Influence of warm ischemia on the onset of tubulointerstitial fibrosis: the percentage of kidney areas displaying interstitial fibrosis stained with Picrosirius was measured over several weeks following warm ischemia within kidneys between W8 and W12 (upper panel). Middle panel: Tubulointerstitial fibrosis determined using Red Picrosirius staining at W12 is shown in lower panel (A, WI-60, B, WI-60-TMZ1 mg, C, WI-60-TMZ5 mg, and D, WI-60-TMZ10 mg). Lower panel: tubular atrophy was determined following the grading described in material and methods section. The values reported are the mean ± SD. *P < .05, **P < .01 WI-60 or WI-60-TMZ1 mg vs WI-60-TMZ5 mg and WI-60-TMZ10 mg. Journal of Vascular Surgery 2008 47, 852-860.e4DOI: (10.1016/j.jvs.2007.10.036) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 6 Influence of warm ischemia on the onset of α-SMA and vimentin expression: Upper panel: α-SMA and vimentin staining in different experimental conditions (A, WI-60, B, WI-60-TMZ1mg, C, WI-60-TMZ5mg, and D, WI-60-TMZ10mg). Middle panel: the percentage of kidney areas displaying α-SMA and vimentin staining with was measured over several weeks following warm ischemia within kidneys between W8 and W12. Lower panel: Tubulointerstitial positive α-SMA cells counted in different experimental groups. The values reported are the mean ±SE. *P < .05, **P < .01 WI-60 or WI-60-TMZ1 mg vs WI-60-TMZ5 mg and WI-60-TMZ10 mg. Journal of Vascular Surgery 2008 47, 852-860.e4DOI: (10.1016/j.jvs.2007.10.036) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions