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- Title
Quantification of glomerular TGF-beta 1 mRNA in patients with diabetes mellitus.
- Authors
Iwano, M; Kubo, A; Nishino, T; Sato, H; Nishioka, H; Akai, Y; Kurioka, H; Fujii, Y; Kanauchi, M; Shiiki, H; Dohi, K
- Abstract
Transforming growth factor-beta 1 (TGF-beta 1) is a primary determinant of the mesangial expansion observed in diabetic nephropathy. In this study, we quantitated the levels of intraglomerular TGF-beta 1 mRNA in patients with diabetes mellitus using a competitive polymerase chain reaction (PCR) method. Renal biopsy specimens were obtained from 29 patients with non-insulin-dependent diabetes mellitus. Total RNA was extracted from the glomeruli and reverse transcribed into cDNA with reverse transcriptase. To prepare samples containing identical amounts of beta-actin cDNA (8 pg), we performed competitive PCR by co-amplifying mutant templates of beta-actin with a unique EcoRI site. We also used this competitive PCR method to measure TGF-beta 1 cDNA by co-amplifying mutant templates of TGF-beta 1. We observed higher expression of TGF-beta 1 mRNA in glomeruli of patients with diabetic nephropathy as compared with normal glomeruli. Intraglomerular TGF-beta 1 mRNA was elevated, even in the early stage of diabetic nephropathy. Moreover, levels of intraglomerular TGF-beta 1 mRNA correlated with values of HbA1c. These data suggest that hyperglycemia induces intraglomerular TGF-beta 1 mRNA expression in vivo, and that TGF-beta 1 overproduction may be associated with the progression of diabetic nephropathy.
- Subjects
RNA analysis; BIOPSY; COLLAGEN; COMPARATIVE studies; DIABETES; DIABETIC neuropathies; DOCUMENTATION; GROWTH factors; IMMUNOBLOTTING; IMMUNOHISTOCHEMISTRY; KIDNEY glomerulus; RESEARCH methodology; MEDICAL cooperation; MICROSCOPY; NUCLEOTIDES; NUCLEOTIDE separation; POLYMERASE chain reaction; RESEARCH; EVALUATION research
- Publication
Kidney International, 1996, Vol 49, Issue 4, p1120
- ISSN
0085-2538
- Publication type
journal article
- DOI
10.1038/ki.1996.162