Cannabinoid receptor 1 blockade ameliorates albuminuria in experimental diabetic nephropathy.
OBJECTIVE
Cannabinoid receptor 1 (CB1) is localized in the central nervous system and in peripheral tissues involved in energy metabolism control. However, CB1 receptors are also expressed at low level within the glomeruli, and the aim of this study was to investigate their potential relevance in the pathogenesis of proteinuria in experimental type 1 diabetes.


METHODS
Streptozotocin-induced diabetic mice were treated with N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,3-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251), a selective CB1-receptor antagonist, at the dosage of 1 mg x kg(-1) x day(-1) via intraperitoneal injection for 14 weeks. Urinary albumin excretion was measured by enzyme-linked immunosorbent assay. CB1 receptor expression was studied by immunohistochemistry, immunoblotting, and real-time PCR. Expression of nephrin, podocin, synaptopodin, and zonula occludens-1 (ZO-1) was assessed by immunofluorescence and real-time PCR. Fibronectin, transforming growth factor-beta1 (TGF-beta1), and connective tissue growth factor (CTGF) mRNA levels were quantitated by real-time PCR.


RESULTS
In diabetic mice, the CB1 receptor was overexpressed within the glomeruli, predominantly by glomerular podocytes. Blockade of the CB1 receptor did not affect body weight, blood glucose, and blood pressure levels in either diabetic or control mice. Albuminuria was increased in diabetic mice compared with control animals and was significantly ameliorated by treatment with AM251. Furthermore, CB1 blockade completely prevented diabetes-induced downregulation of nephrin, podocin, and ZO-1. By contrast overexpression of fibronectin, TGF-beta1, and CTGF in renal cortex of diabetic mice was unaltered by AM251 administration.


CONCLUSIONS
In experimental type 1 diabetes, the CB1 receptor is overexpressed by glomerular podocytes, and blockade of the CB1 receptor ameliorates albuminuria possibly via prevention of nephrin, podocin, and ZO-1 loss.
Barutta
Federica
F
Diabetic Nephropathy Laboratory, Department of Internal Medicine, University of Turin, Turin, Italy.
Corbelli
Alessandro
A
Mastrocola
Raffaella
R
Gambino
Roberto
R
Di Marzo
Vincenzo
V
Pinach
Silvia
S
Rastaldi
Maria Pia
MP
Perin
Paolo Cavallo
PC
Gruden
Gabriella
G
D000419:Albuminuria / Q000517:prevention & control*; D000818:Animals; D055513:Connective Tissue Growth Factor / Q000235:genetics; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000150:complications / Q000503:physiopathology; D003928:Diabetic Nephropathies / Q000517:prevention & control*; D005353:Fibronectins / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D010750:Phosphoproteins / Q000235:genetics; D010880:Piperidines / Q000627:therapeutic use; D016133:Polymerase Chain Reaction / Q000379:methods; D011720:Pyrazoles / Q000627:therapeutic use; D012333:RNA, Messenger / Q000235:genetics; D043884:Receptor, Cannabinoid, CB1 / Q000037:antagonists & inhibitors*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053773:Transforming Growth Factor beta1 / Q000235:genetics; D062826:Zonula Occludens-1 Protein
C526318:CCN2 protein, mouse; D005353:Fibronectins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C407223:NPHS2 protein; D010750:Phosphoproteins; D010880:Piperidines; D011720:Pyrazoles; D012333:RNA, Messenger; D043884:Receptor, Cannabinoid, CB1; C569043:Tjp1 protein, mouse; D053773:Transforming Growth Factor beta1; D062826:Zonula Occludens-1 Protein; C113267:nephrin; D055513:Connective Tissue Growth Factor; C103505:AM 251