Effect of nitric oxide-cGMP-dependent protein kinase activation on advanced glycation end-product-induced proliferation in renal fibroblasts.
Renal interstitial fibrosis is believed to play a key role in the development of diabetic nephropathy (DN), and advanced glycation end-products (AGE) may contribute importantly to this. Recent reports have shown that nitric oxide (NO) is closely linked to the renal interstitial fibrosis of DN. In this study, the mechanisms by which NO and its downstream signals mediate the AGE-induced proliferative response in normal rat kidney fibroblasts (NRK-49F) are examined. AGE decreased NO production, cyclic guanosine 5'monophosphate (cGMP) synthesis, and cGMP-dependent protein kinase (PKG) activation time- and dose-dependently. These effects were not observed when cells were treated with nonglycated BSA. NO and inducible nitric oxide synthase (iNOS) stimulated by NO donors S-nitroso-N-acetylpenicillamine (SNAP)/sodium nitroprusside (SNP) and PKG activator 8-para-chlorophenylthio-cGMP (8-pCPT-cGMP) prevented both AGE-induced proliferation and Janus kinase 2 (JAK2)-signal transducers and activators of transcription 5 (STAT5) activation but not p42/p44 mitogen-activated protein kinase (MAPK) activation. The ability of NO-PKG to inhibit AGE-induced cell cycle progression was verified by the observation that SNAP, SNP, and 8-pCPT-cGMP inhibited both cyclin D1 and cdk4 activation. Furthermore, induction of NO-PKG significantly increased p21Waf1/Cip1 expression in AGE-treated NRK-49F cells. The data suggest that the NO-PKG pathway inhibits AGE-induced proliferation by suppressing activation of JAK2-STAT5 and cyclin D1/cdk4 and induction of p21Waf1/Cip1.
Huang
Jau-Shyang
JS
Department of Biological Science and Technology, Chung Hwa College of Medical Technology, 89 Wen-Hwa, 1st Street, Jen-Te Hsiang, Tainan Hsien 717, Taiwan, Republic of China. jaushyang12@hotmail.com
Chuang
Lea-Yea
LY
Guh
Jinn-Yuh
JY
Chen
Chorng-Jia
CJ
Yang
Yu-Lin
YL
Chiang
Tai-An
TA
Hung
Min-Yuan
MY
Liao
Tung-Nan
TN
D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism*; D056741:Cyclin D; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism; D003928:Diabetic Nephropathies / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D047468:Immunoprecipitation; D053614:Janus Kinase 2; D007668:Kidney / Q000166:cytology*; D008954:Models, Biological; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D009599:Nitroprusside / Q000494:pharmacology; D009841:Oligonucleotides / Q000737:chemistry; D018384:Oxidative Stress; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051381:Rats; D026423:S-Nitroso-N-Acetylpenicillamine / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D013873:Thionucleotides / Q000378:metabolism; D013997:Time Factors
C493958:Cdkn1a protein, rat; D056741:Cyclin D; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D017127:Glycation End Products, Advanced; D009841:Oligonucleotides; D011518:Proto-Oncogene Proteins; C110881:S-nitro-N-acetylpenicillamine; D050799:STAT5 Transcription Factor; D013873:Thionucleotides; D009599:Nitroprusside; D009569:Nitric Oxide; C055656:8-((4-chlorophenyl)thio)cyclic-3',5'-GMP; D026423:S-Nitroso-N-Acetylpenicillamine; D052247:Nitric Oxide Synthase Type II; D011505:Protein-Tyrosine Kinases; C507927:Jak2 protein, rat; D053614:Janus Kinase 2; D017869:Cyclic GMP-Dependent Protein Kinases; D051358:Cyclin-Dependent Kinase 4; D010396:Penicillamine; D006152:Cyclic GMP