Figures & data
Figure 1. Effect of diabetes on biochemical parameters in vivo. Diabetes increased levels of blood sugar (A), HbA1c (B), kidney/body weight (%) (D) and urinary protein/Cr (E) in experimental animals. Diabetes mildly but no significantly attenuated body weight (C) and no significantly difference in serum Cr level (F) between these two groups. Rats were given STZ to induce diabetes. Symbol * indicates significant difference from the normal group (p < 0.05).
Figure 2. (A) Immunohistochemical photograph of glomeruli in rats with and without diabetes. The diabetic group expressed stronger type I collagen (A) and type IV collagen (B) expression when compared with the normal group. Semi-quantitative evaluation of the positive immune-labeled cells was shown below for type I and IV collagen staining. Data were expressed as mean ± SE. *p < 0.05 versus control group. Specimens were observed under magnification 400×. (C) Histological examination of rat glomeruli subjected to magnetic bead perfusion. An adult rat was perfused with magnetic beads through the heart and glomeruli were isolated.
Figure 3. (A) Representative 2D gel electrophoretograms of glomerular proteins in rats with and without diabetes; 250 μg of glomerular lysate was subjected to IEF (pH 4–7), SDS–polyacrylamide gel separation and silver staining. (B) Enlarged regions of the 10 spots of interest in the silver-stained SDS-polyacrylamide gels. The spots in the gels of normal glomeruli and diabetic glomeruli from rats were matched using Bio-Rad Proteoweaver 2D Analysis Software Version 4.0. The arrows in the images indicate the spots of interest in both groups. (C) Representative mass spectrum of Spot 15863 (Protein disulfide isomerase associated 3). In-gel trypsin digestion products were assessed using an Ultraflex™ TOF/TOF mass spectrophotometer. Peptide mass fingerprints were submitted to the NCBI or the SwissPort bioinformation stations using MASCOT search engine. The X- and Y-axes in the mass spectrum indicate m/z and intensities of peptides (arbitrary unit × 104), respectively.
Table 1. The mass spectrometric identified proteins.
Figure 4. Relative intensities of the positively identified proteins. Rats with diabetic glomeruli displayed higher levels of the (A) protein disulfide isomerase associated 3, (B) aspartoacylase-3, (C) 3-hydroxymethyl-3-methylglutaryl-Coenzyme A lyase, (D) lactamase beta 2 and (E) Agmat protein (171.2 ± 76.7%; p < 0.01). The intensity of protein disulfide isomerase associated 3 remained undetectable in six subjects of the normal group. The relative density was calculated by dividing the density of matched spot by the density of all the matched spots in the respective gel. * indicates significant difference from the normal group, p < 0.05.
Figure 5. Rats with diabetic isolated glomeruli had lower levels of (A) regucalcin, (B) rCG52140, (C) aldo-keto reductase family 1, (D) peroxiredoxin 1, and (D) l-arginine: glycine and (E) amidinotransferase. The relative density was calculated by dividing the density of matched spot by the density of all the matched spots in the respective gel. * indicates significant difference from the normal group, p < 0.05.
