Attenuation of collagen deposition by sarpogrelate and rosuvastatin.

<p>(A) Kidney tissues were stained with Sirius red and cortex areas with collagen deposition were quantified. Two histological fields for each kidney tissue were analyzed. All data represent means ± SD. Scale bar = 120 μm (B) Protein levels of collagen type IV in kidney homogenates were assessed with western blot analysis. Data are means ± SD from three determinants of pooled samples. ^aP < 0.05 compared with the NFD control group. ^bP < 0.05 compared with the HFD/STZ group. ^cP < 0.05 compared with the HFD/STZ +Sa group. ^dP < 0.05 compared with the HFD/STZ + R group. (C) Protein levels of renal collagen type Ia1 were monitored with western blot analysis. Similar blots were obtained from three independent blots.</p

Effects of sarpogrelate and rosuvastatin on histomorphological changes in HFD/STZ mice.

<p>(A) H&E staining was performed in kidney tissues and representative histopathological images (cortex (a), glomerulus (b), and renal tubules (c)) are shown. Scale bar = 120 μm. (B) Numbers of glomeruli that display vasodilation and mesangial expansion were quantified with a computer-based automated image analyzer. (C) Numbers of tubules with vacuolation were quantified. (D) Oil red O staining was conducted and cortex areas with positive staining were quantified with a computer-based automated image analyzer. (E) Immunohistochemical staining was performed with caspase-3-specific antibody. Areas with caspase-3-positive staining were quantified. ^aP < 0.05 compared with the NFD control group. ^bP< 0.05 compared with the HFD/STZ group. ^cP < 0.05 compared with the HFD/STZ + Sa group. ^dP < 0.05 compared with the HFD/STZ + R group. Two histological fields for each kidney tissue were analyzed. All data represent means ± SD. Scale bar = 120 μm.</p

Suppressive effects of sarpogrelate on TGF-β1/5-HT-mediated PAI-1 increase in mesangial cells.

<p>(A) MES13 cells were incubated with sarpogrelate (Sa, 10 μM) along with vehicle (V, medium) or rosuvastatin (R, 2.5–40 μM) for 24 h, and viable cell numbers were assessed with MTT analysis. Data represent ratios with respect to untreated (UT) control group. Data are means ± SD from three experiments. (B) MES13 cells were pre-incubated with sarpogrelate (10 μM) for 6 h and then TGF-β1 (10 ng/ml) was additionally incubated for 24 h along with either vehicle (V) or 5-HT (10 μM). Levels of PAI-1 were assessed with western blotting. Data are means ± SD from three independent experiments. (C) MES13 cells were pretreated with sarpogrelate (10 μM), rosuvastatin (10 μM) or both for 6 h, and then TGF-β1 (10 ng/ml) and 5-HT (10 μM) were co-incubated for a further 24 h. Data are means ± SD from three independent experiments. (D) After the treatment of MES13 with sarpogrelate and TGF-β1/5-HT, transcript levels for PAI-1 were quantified with relative real-time RT-PCR analysis. Sarpogrelate was pre-incubated for 6 h and the TGF-β1/5-HT incubation was followed for a further 24 h. Data are means ± SD from three experiments.</p

Inhibitory effects of sarpogrelate and rosuvastatin on pro-fibrotic factor TGF-β1 and PAI-1.

<p>(A-B) Immunohistological analysis was performed with TGF-β1 and PAI-1 antibodies. Cortex areas with TGF-β1 (A) and PAI-1 (B) expression were quantified. Two histological fields for each kidney tissue were analyzed. All data represent means ± SD. Scale bar = 120 μm. (C) Protein levels of PAI-1 in the kidney were determined with western blot analysis. Data are means ± SD from 3 measurement of pooled samples. ^aP < 0.05 compared with NFD control group. ^bP < 0.05 compared with the HFD/STZ group. ^cP < 0.05 compared with the HFD/STZ + Sa group. ^dP < 0.05 compared with the HFD/STZ + R group. (D) Serum levels of PAI-1 were monitored in blood samples that were obtained at the terminal step using an ELISA-based kit. Data are means ± SD from 4–8 samples. ^aP < 0.05 compared with the NFD control group.</p

Amelioration effects of sarpogrelate and rosuvastatin on the expression of angiogenic markers CD31 and VEGFR-2.

<p>(A) Immunohistochemcial analyses were carried out with CD31- or VEGFR-2-specific antibodies and representative immunoreactive cortex regions are shown. (B-C) The CD31 (B) or VEGFR-2 (C)-positive cortex areas were quantified. ^aP < 0.05 compared with the NFD control group. ^bP< 0.05 compared with the HFD/STZ group. ^cP < 0.05 compared with the HFD/STZ + Sa group. ^dP < 0.05 compared with the HFD/STZ + R group. Two histological fields for each kidney tissue were analyzed. All data represent means ± SD. Scale bar = 120 μm.</p

After the completion of animal experiment, body weight and kidney weight were determined, and blood levels of cholesterol, triglycerides, and BUN were measured.

<p>Data are means ± SD</p

Establishment of the HFD/STZ mouse model.

<p>(A) After STZ treatment at Week 8, body weight changes in HFD-fed mice were monitored once per week. HFD was stared a Week 1 and was continued for 22 weeks. Drug treatments began at Week 9 and continued for 13 weeks. (B) Fasting blood glucose levels were monitored at Week 15 of HFD. Mice were fasted overnight (12 h) and blood glucose levels were measured with an Accu-Chek blood glucose meter. The data are means ± standard deviation (SD) from 4–8 samples. ^aP < 0.05 compared with the NFD group.</p

Effects of sarpogrelate (Sa) and rosuvastatin (R) on HFD/STZ-induced renal function.

<p>(A) At Week 20 of the HFD, urine samples were collected overnight (15 h) for three consecutive days, and levels of urine albumin were determined in pooled urines. Albumin levels were normalized with corresponding urine creatinine levels to obtain the urine albumin-to-creatinine ratio (UACR) values. (B) Urine cystatin C levels were normalized with corresponding creatinine (CR) levels. The data are means ± SD from 3 pooled urine samples. ^aP < 0.05 compared with the NFD group. ^bP < 0.05 compared with the HFD/STZ group.</p

A 38-year-old woman with invasive ductal carcinoma (tumor size 37 mm, triple negative, strongly positive Ki-67).

<p>The HR-MAS MR spectrum (11.7T) obtained using the core needle biopsy specimen shows peaks of each choline metabolite. The tCho concentration measured with HR-MAS MR spectroscopy was 6.5 mmol/kg. Note. Lac, lactate; Ala, alanine; Glu, glutamate; Cr, creatine; Cho, free choline; GPC, glycerophosphocholine; PC, phosphocholine; tCho, total choline, sum of Cho, PC, and GPC; Tau, taurine; m-Ins, myo-inositol; Gly, glycine.</p

HR-MAS MR spectroscopy values for 36 breast cancer specimens.

<p>Data represent the mean ± standard deviation (median).</p><p>Cho: choline, PC: phosphocholine, GPC: glycerophosphocholine, tCho: total choline (the sum of Cho, PC, and GPC), Cr: creatine, Tau: taurine, Gly: glycine, m-Ins: myo-inositol, s-Ins: scyllo-inositol, Ala: alanine, Suc: succinate.</p