Open AccessDetection of glycosylated protein in glomeruli of STZ‐induced diabetic rats using the nitroblue tetrazolium (NBT) reaction
Author(s) -
Saitoh Akemi,
Tomino Yasuhiko,
Kuramoto Takao,
Fukui Mitsumine,
Ohmuro Hiroyuki,
Shirato Isao,
Koide Hikaru,
Itoh Kiichi
Publication year - 1994
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860080404
Subject(s) - fructosamine , diabetic nephropathy , medicine , endocrinology , laminin , fibronectin , streptozotocin , diabetes mellitus , immunofluorescence , chemistry , type iv collagen , extracellular matrix , immunology , biochemistry , antibody
Detection of glycosylated protein in renal tissues was determined in streptozotocin (STZ)‐induced diabetic rats using the nitroblue tetrazolium (NBT) reaction. The glycosylation of extra‐cellular matrix (ECM) components such as laminin and fibronectin was examined in vitro using the same method. Immunofluorescence staining of laminin or type IV collagen was also performed in renal tissues of STZ‐induced diabetic rats. There was no significant difference in the intensity of NBT in renal tissues between 4 week STZ‐induced diabetic rats and control rats of the same age. The intensity of NBT staining in the glomerular mesangial areas and capillary walls was marked in 12 week diabetic rats. The mean values of fructosamine measured by the NBT reaction in the glycosylated‐laminin and fibronectin were increased dose dependently. In immunofluorescence, laminin and type IV collagen were observed significantly in the glomerular mesangial areas and capillary walls of 12 week diabetic rats. However, there was no significant change in renal histopathology in 4 and 12 week diabetic rats. It appears that the non‐enzymatic glycosylation and expression of ECM components in glomeruli increased in the early stage of diabetic nephropathy prior to the appearance of marked histologic alterations. In conclusion, non‐enzymatic glycosylation of glomerular structural components may play an important role in the initiation of the early stage of renal injuries in diabetes. © 1994 Wiley‐Liss, Inc.