title: The Notch pathway mediates the angiotensin II-induced synthesis of extracellular matrix components in podocytes.
abstract: The Notch pathway is known to contribute to the development of glomerular disease. Angiotensin II (Ang II), an important member of the renin-angiotensin system, stimulates the accumulation of extracellular matrix components in glomerular disease; however, the exact mechanisms involved remain to be elucidated. In the present study, we aimed to investigate the effects of the Notch pathway on the synthesis of extracellular matrix components in Ang II-stimulated podocytes. Mouse podocytes were stimulated with Ang II (10-6 mol/l). The activation of the Notch pathway was inhibited by a vector carrying short hairpin RNA (shRNA) targeting Notch1 (sh-Notch1) or by γ-secretase inhibitor (GSI). The protein levels of Notch1, Notch intracellular domain 1 (NICD1), hairy and enhancer of split-1 (Hes1), matrix metalloproteinase (MMP)-2, MMP-9, transforming growth factor-β1 (TGF-β1), type IV collagen and laminin were determined by western blot analysis. The Notch1, Hes1, MMP-2, MMP-9, TGF-β1, type IV collagen and laminin mRNA levels were detected by RT-PCR. The MMP-2 and MMP-9 activity was measured using a cell active fluorescence assay kit. The levels of TGF-β1, type IV collagen and laminin were determined in the culture medium of the podocytes by enzyme-linked immunosorbent assay (ELISA). Our results revealed that Ang II upregulated Notch1, NICD1, Hes1, TGF-β1, type IV collagen and laminin expression and downregulated MMP-2 and MMP-9 expression in the cultured podocytes. The inhibition of the Notch pathway by sh-Notch1 or GSI increased MMP-2 and MMP-9 expression, decreased the TGF-β1 level and suppressed type IV collagen and laminin expression. The inhibition of the Notch pathway by sh-Notch1 or GSI also increased MMP-2 and MMP-9 activity, and decreased TGF-β1 levels, type IV collagen levels and laminin secretion. These findings indicate that the Notch pathway potentially mediates the Ang II-induced synthesis of extracellular matrix components in podocytes through the regulation of MMPs and TGF-β1.
authors.affiliation: Department of Pathology, Τhe Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Yao
authors.affiliation: Department of Pathology, Τhe Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Pathology, Τhe Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Nephrology, Τhe Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Department of Nephrology, Τhe Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Yanqing
authors.initials: Y
authors.lastname: Chi
authors.affiliation: Department of Pathology, Τhe Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors; D000804:Angiotensin II / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D002460:Cell Line; D024141:Collagen Type IV / Q000378:metabolism; D004789:Enzyme Activation; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis*; D018398:Homeodomain Proteins / Q000378:metabolism; D007678:Kidney Glomerulus / Q000503:physiopathology; D007797:Laminin / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D009842:Oligopeptides / Q000494:pharmacology; D050199:Podocytes / Q000166:cytology / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D051881:Receptor, Notch1 / Q000235:genetics / Q000378:metabolism*; D000072056:Transcription Factor HES-1; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*
chemical_list: D053829:Amyloid Precursor Protein Secretases; D000804:Angiotensin II; D051792:Basic Helix-Loop-Helix Proteins; D024141:Collagen Type IV; D016326:Extracellular Matrix Proteins; D018398:Homeodomain Proteins; D007797:Laminin; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D009842:Oligopeptides; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051881:Receptor, Notch1; D000072056:Transcription Factor HES-1; D053773:Transforming Growth Factor beta1; C489734:Hes1 protein, mouse; C497762:Notch1 protein, mouse; C523462:benzyloxycarbonyl-leucyl-leucyl-norleucinal; C522362:Mmp2 protein, mouse; C501830:Mmp9 protein, mouse