Histological and Functional Assessment of Diabetic Nephropathy

Kidney sections were stained with Periodic-Acid Schiff (PAS) and Masson’s trichrome to reveal renal morphologic changes^{23 (link),48 (link)}. The changes of DN features—glomerulosclerosis (based on PAS images, scale from 0 to 4) and glomerular fibrosis (based on Masson staining) were scored with the scorer blinded to the group^{23 (link),48 (link)}. Relative staining was quantified with NIH Image J software (Bethesda, MD, USA). The images (N = 6~10 per animal, 6–11 mice/group) were analyzed and quantitated in a blinded fashion^{23 (link),48 (link)}. Glomerular filtration rate (GFR) was measured in conscious mice by the fluorescein isothiocyanate-inulin method as reported previously^{23 (link),49 (link)}, as recommended by the Diabetic Complications Consortium (http://www.diacomp.org/). Urine samples, collected from mice individually housed in metabolic cages, were assayed for albumin/creatinine ratio (ACR) (Albuwell and Creatinine Companion, Exocell Inc., Philadelphia, PA, USA)^{23 (link),49 (link)}.

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Zhao X.P., Chang S.Y., Liao M.C., Lo C.S., Chenier I., Luo H., Chiasson J.L., Ingelfinger J.R., Chan J.S, & Zhang S.L. (2018). Hedgehog Interacting Protein Promotes Fibrosis and Apoptosis in Glomerular Endothelial Cells in Murine Diabetes. Scientific Reports, 8, 5958.

Publication 2018

Albuwell and Creatinine Companion

Albumin Animal Companion Conscious Creatinine Diabetic complications Fibrosis Fluorescein Glomerular Glomerular filtration rate Inulin Isothiocyanate Mice Periodic acid Renal Urine

Corresponding Organization : Centre Hospitalier de l’Université de Montréal

Other organizations : Massachusetts General Hospital, Harvard University

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Variable analysis

independent variables

Kidney sections were stained with Periodic-Acid Schiff (PAS) and Masson's trichrome to reveal renal morphologic changes

dependent variables

Glomerulosclerosis (based on PAS images, scale from 0 to 4)
Glomerular fibrosis (based on Masson staining)
Relative staining quantified with NIH Image J software
Glomerular filtration rate (GFR) measured in conscious mice by the fluorescein isothiocyanate-inulin method
Urine albumin/creatinine ratio (ACR)

control variables

Scorer blinded to the group

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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