Suppression of nNOS expression in rat enteric neurones by the receptor for advanced glycation end-products.
Diabetes mellitus results in a loss of neuronal nitric oxide synthase (nNOS) expression in the myenteric plexus but the underlying mechanisms remain unknown. We hypothesized that this may be mediated by advanced glycation end-products (AGEs), a class of modified protein adducts formed by non-enzymatic glycation that interact with the receptor for AGE (RAGE) and which are important in the pathogenesis of other diabetic complications. Whole mount preparations of longitudinal muscles with adherent myenteric plexus (LM-MPs) from the duodenum of adult male rats were exposed to glycated bovines serum albumin (AGE-BSA) or BSA for 24 h. Western blotting, immunohistochemistry and real-time reverse transcriptase polymerase chain reaction (RT-PCR) for mRNA showed a significant reduction in nNOS expression in LM-MPs after exposure to AGE-BSA. NO release, as measured by the Griess reaction, was also significantly reduced by AGE-BSA. A neutralizing antibody against RAGE attenuated the reduction of nNOS protein caused by AGE-BSA. Immunohistochemistry revealed co-localization of RAGE expression with Hu, a marker for neuronal cells but not for S-100, a glial marker. Advanced glycation end-products reduce nNOS expression in the rat myenteric neurones acting via the receptor RAGE. Our results suggest novel pathways for disruption of the nitrergic phenotype in diabetes.
Korenaga
K
K
Enteric Neuromuscular Disorders and Pain Group, Division of Gastroenterology and Hepatology, Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX 77555-0764, USA.
Micci
M-A
MA
Taglialatela
G
G
Pasricha
P J
PJ
D000818:Animals; D015153:Blotting, Western; D003920:Diabetes Mellitus / Q000503:physiopathology; D004386:Duodenum / Q000294:innervation / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D009130:Muscle, Smooth / Q000294:innervation / Q000378:metabolism; D009197:Myenteric Plexus / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D052248:Nitric Oxide Synthase Type I / Q000096:biosynthesis*; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D052248:Nitric Oxide Synthase Type I