[Frontiers in Bioscience 11, 968-976, January 1, 2006] 968 Chronically increased intrarenal angiotensin II causes nephropathy in an animal model of type 2 diabetes

1. Abstract 2. Introduction 3. Methods 3.1. Animals 3.2. Urine Collection 3.3. Blood collection and kidneys removal 3.4. Determination of plasma and intra-renal ANG II 3.4.1. Extraction of ANG peptides by Solid Phase Extraction Chromatography (SPEC) 3.4.2. Quantification of ANG II levels by radioimmunoassay 3.5. Measurement of functional parameters 3.6. Isolation of glomeruli 3.6.1. Preparation of glomerular RNA 3.6.2. Preparation of glomerular protein lysate 3.7. Assessment of changes in the levels of mRNA for nephrin by RT-PCR 3.8. Assessment of glomerulosclerosis 3.8.1. Assessment of changes in the levels of mRNA for collagen types IV and fibronectin by RT-PCR 3.8.2. Assessment of changes in protein levels of collagen types IV and fibronectin 3.8.3. Histology 3.9. Statistics 4. Results 4.1. Measurement of body weight, mean arterial blood pressure, serum insulin and glucose 4.2. Measurement of proteinuria 4.3. Measurement of plasma and intra-renal ANG II 4.4. Expression of nephrin by RT-PCR 4.5. Documentation of sclerotic changes 5. Discussion 6. Acknowledgment 7. References