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Effects of advanced glycation end products on renal fibrosis and oxidative stress in cultured NRK-49F cells
- Source :
- Chinese medical journal. 120(9)
- Publication Year :
- 2007
-
Abstract
- Background Advanced glycation end products (AGEs) play a critical role in the development of diabetic nephropathy. Reactive oxygen species (ROS) may play a critical role in AGEs induced growth factor expression. In this study, the effects of AGEs on transforming growth factor beta1 (TGF-beta1), connective tissue growth factor (CTGF) and fibronectin (Fn) mRNA expression and oxidative stress in cultured NRK-49F cells were examined. Methods NRK-49F cells were incubated with medium containing different doses of AGEs (50, 100 or 200 microg/ml) for 24 hours, or with AGEs 100 microg/ml for different times (0, 12, 24 or 48 hours). Cells in the serum-free medium or medium containing 25 mmol/L glucose were controls. Cells were treated with 25 mmol/L glucose and 100 microg/ml AGEs for 24 hours to determine the effects between AGEs and glucose. We clarified the role of antioxidant by pretreating cells with N-acetylcysteine (10 mmol/L), ginkgo biloba extract (50 or 100 mg/L) for 24 hours and with 100 microg/ml AGEs for further 24 hours. Alamarblue dye assay was used to analyze cell growth; intracellular ROS generation was measured by flow cytometry; intracellular glutathione by fluorescence spectrophotometry; expressions of TGF-beta1, CTGF and Fn mRNA by semiquantitative RT-PCR. Results AGEs significantly increased the expressions of TGF-beta1, CTGF, Fn mRNA and intracellular ROS generation, and decreased the glutathion level in NRK-49F cells in dose- and time-dependent manners. High glucose and AGEs together significantly increased the expression of TGF-beta1, CTGF and Fn mRNA, compared with AGEs and high glucose separately. Preincubation with N-acetylcysteine or ginkgo biloba extract increased GSH level, suppressed AGEs-induced oxidative stress and TGF-beta1, CTGF and Fn mRNA overexpression. Conclusions AGEs can significantly increase expression of TGF-beta1, CTGF, Fn mRNA in NRK-49F cells through enhancement of oxidative stress. The accumulation of AGEs may play a pivotal role in the pathogenesis of tubulointerstitial fibrosis in diabetic nephropathy. Suppression of AGEs induced TGF-beta1, CTGF and Fn mRNA overexpression in renal fibroblasts through inhibition of oxidative stress may be a mechanism underlying effect of ginkgo biloba extract in diabetic nephropathy. In addition, antioxidant therapy may help prevent AGEs accumulation and its induced damage.
- Subjects :
- Glycation End Products, Advanced
medicine.medical_specialty
medicine.medical_treatment
medicine.disease_cause
Kidney
Immediate early protein
Antioxidants
Immediate-Early Proteins
Transforming Growth Factor beta1
chemistry.chemical_compound
Glycation
Fibrosis
Internal medicine
medicine
Humans
Diabetic Nephropathies
Cells, Cultured
Cell Proliferation
Chemistry
Growth factor
Connective Tissue Growth Factor
Kidney metabolism
General Medicine
Glutathione
medicine.disease
Fibronectins
CTGF
Oxidative Stress
Endocrinology
Intercellular Signaling Peptides and Proteins
Oxidative stress
Subjects
Details
- ISSN :
- 03666999
- Volume :
- 120
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- Chinese medical journal
- Accession number :
- edsair.doi.dedup.....bd09cae4f2dcb7460daedde74cae420b