mmp 2 inhibitor sb 3ct  (MedChemExpress)

Journal: Kidney International Reports

Article Title: COL4A3 Mutation Induced Podocyte Apoptosis by Dysregulation of NADPH Oxidase 4 and MMP-2

doi: 10.1016/j.ekir.2023.06.007

Figure Lengend Snippet: Characterization of C ol4a3 transgenic mouse model. (a) Western blot analysis of type IV collagen α3 chain level in glomeruli of C ol4a3 transgenic mice and wild type (WT) mice. The transgenic group (Mut) had a significantly lower type IV collagen α3 chain level compared with the WT group. (b) Sanger sequencing of C ol4a3 mutant mice constructed by CRISPR/Cas9 technology. (c)–(e) The mutant and WT mice treated with or without NOX4 inhibitor at 28 weeks of age, quantification of urinary ACR ([c] each group N = 4), 24 hour proteinuria ([d] each group N = 3), serum creatinine ([e] each group N = 3, and serum creatinine was transformed as Ln[Scr] because of skewed distribution). ∗∗ P < 0.05, ∗∗∗ P < 0.001. (f) Real-time PCR validation show the expression of NOX4 and MMP-2 mRNA was higher in mutant group compared with that in WT group. N = 4; ∗∗ P < 0.01. (g) Representative light micrographs of HE stained kidney sections from WT and mutant mice at 28 weeks of age. Light micrographs represent focal adhesion of glomeruli (red arrow), tubular cast (yellow arrow), interstitial inflammatory cell infiltration (yellow triangle) in mutant group. N = 4. (h) Electron micrographs of kidney tissue from WT and mutant mice at 28 weeks of age. Electron micrographs show partial podocyte foot process effacement (red star), segmental thickening of GBM (red arrow) in mutant group. N = 4. ACR, albumin-to-creatinine ratio.

Article Snippet: MMP-2 inhibitor, SB-3CT (Selleckchem, USA) was administrated intraperitoneally for 6 days before sacrifice at the dosage of 50 mg/kg; NOX4 inhibitor, GKT137831 (Selleckchem, USA) was administrated gavage for 28 days before sacrifice at the dosage of 60 mg/kg.

Techniques: Transgenic Assay, Western Blot, Sequencing, Mutagenesis, Construct, CRISPR, Transformation Assay, Real-time Polymerase Chain Reaction, Expressing, Staining

Journal: Kidney International Reports

Article Title: COL4A3 Mutation Induced Podocyte Apoptosis by Dysregulation of NADPH Oxidase 4 and MMP-2

doi: 10.1016/j.ekir.2023.06.007

Figure Lengend Snippet: Microarray analysis of patients with ADAS with COL4A3 mutation followed by validation of DEGs from both transcriptomic and proteomic level. (a) The heat map of gene expression analysis in glomeruli of patients with ADAS and patients with MCD. The line highlighted represents top highest DEG, MMP-2. Red indicates upregulated. Green indicates down-regulated. Black indicates no significant difference between the 2 groups. F1–F3 indicated patients with ADAS with COL4A3 mutation. M1–M5 indicates patients with MCD as control. (b)–(c) Pathways of genes upregulated in patients with ADAS compared with patients with MCD, using Kyoto Encyclopedia of Genes and Genomes analysis (b), and Enrichr Gene Ontology Biological Process program (c). (d) Real-time PCR validation of NOX4 and MMP-2 in glomeruli of 3 additional patients with ADAS and 7 additional patients with MCD. (e) Immunohistochemical staining of MMP-2 in glomeruli of patients with ADAS and patients with MCD. (f) Histogram of immunohistochemical staining from (e) showed a significant increase in the expression of MMP-2 in glomeruli of patients with ADAS compared with that in glomeruli of patients with MCD. ∗ P < 0.05, ∗∗ P < 0.01. N = 3. ADAS, autosomal dominant Alport syndrome; MCD, minimal change disease.

Article Snippet: MMP-2 inhibitor, SB-3CT (Selleckchem, USA) was administrated intraperitoneally for 6 days before sacrifice at the dosage of 50 mg/kg; NOX4 inhibitor, GKT137831 (Selleckchem, USA) was administrated gavage for 28 days before sacrifice at the dosage of 60 mg/kg.

Techniques: Microarray, Mutagenesis, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemistry, Staining

Journal: Kidney International Reports

Article Title: COL4A3 Mutation Induced Podocyte Apoptosis by Dysregulation of NADPH Oxidase 4 and MMP-2

doi: 10.1016/j.ekir.2023.06.007

Figure Lengend Snippet: Validation of the relation between NOX4 and MMP-2. (a) In vitro validation, Western blot analysis of NOX4, MMP-2, and apoptosis-related protein cleaved caspase 3 in COL4A3 transgenic podocytes. (b) In vivo analysis, Western blot verification showed increased expression of NOX4, MMP-2, and cleaved caspase 3 in glomeruli of C ol4a3 transgenic mouse. (c) Measurement of podocyte H 2 O 2 before and after NOX4 inhibition. ( N = 4). (d) Densitometric analysis represented Western blot analysis from (a). ( N = 4) (e) Densitometric analysis represented Western blot analysis from (b). ( N = 4) ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: MMP-2 inhibitor, SB-3CT (Selleckchem, USA) was administrated intraperitoneally for 6 days before sacrifice at the dosage of 50 mg/kg; NOX4 inhibitor, GKT137831 (Selleckchem, USA) was administrated gavage for 28 days before sacrifice at the dosage of 60 mg/kg.

Techniques: In Vitro, Western Blot, Transgenic Assay, In Vivo, Expressing, Inhibition

Journal: Bioscience Reports

Article Title: Resveratrol-Linoleate protects from exacerbated endothelial permeability via a drastic inhibition of the MMP-9 activity

doi: 10.1042/BSR20171712

Figure Lengend Snippet: After seeding primary HUVEC (3 × 10 5 ) on collagen-coated insert, cells were incubated for 48 h. Different concentrations of NADES or DMSO incubated for 24 h at 37°C with primary HUVEC monolayers ( A ). Cells were incubated with TNF-α 100 ng/ml with or without 10 µM of RES, RES-LA, or SB-3CT ( B ). Data represent mean ± S.D.; ** P ≤0.01, *** P ≤0.001.

Article Snippet: SB-3CT (commercial MMP-2, -9 inhibitor) was purchased from Enzo Life Sciences™ (Villeurbanne, France).

Techniques: Incubation

Journal: Bioscience Reports

Article Title: Resveratrol-Linoleate protects from exacerbated endothelial permeability via a drastic inhibition of the MMP-9 activity

doi: 10.1042/BSR20171712

Figure Lengend Snippet: HUVEC (3 × 10 5 cells/well) were plated for 48 h on biotinylated-gelatin coated chambers. TNF-α (100 ng/ml) was added in the presence or in the absence of 10 µM RES, RES-LA, SB-3CT, or with anti-MMP-9 mAb for 24 h. All samples were treated either with a Cy3-conjugated mouse anti-human CD31/PECAM-1 mAb ( A ) or with a fluorescein-streptavidin compound ( B ).

Article Snippet: SB-3CT (commercial MMP-2, -9 inhibitor) was purchased from Enzo Life Sciences™ (Villeurbanne, France).

Techniques:

Journal: Bioscience Reports

Article Title: Resveratrol-Linoleate protects from exacerbated endothelial permeability via a drastic inhibition of the MMP-9 activity

doi: 10.1042/BSR20171712

Figure Lengend Snippet: HUVEC were seeded (2 × 10 5 cells/well) in gelatin A-coated golden ECIS ‘8W10E+’ well arrays. After formation of a confluent HUVEC monolayer, cells were treated with 100 ng/ml TNF-α in the presence or absence of 10 µM of either RES, RES-LA, or SB-3CT. Real-time TEER measurements were done at and 8-s frequency. Results were analyzed using ECIS software. ( A ) Represents untreated, negative controls (0.1% NADES or DMSO), and TNF-α groups. ( B ) Represents RES, RES-LA, or SB-3CT groups.

Article Snippet: SB-3CT (commercial MMP-2, -9 inhibitor) was purchased from Enzo Life Sciences™ (Villeurbanne, France).

Techniques: Software

Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors

doi: 10.1007/978-1-4939-7111-4_8

Figure Lengend Snippet: Representative synthetic MMPIs and their IC 50 or K i toward specific MMPs.

Article Snippet: For example, stock solution (10 −2 M) of SB-3CT (MMP-2/MMP-9 inhibitor IV) (EMD Millipore), Ro-28-2653 (5-biphenyl-4-yl-5-[4-(-nitro-phenyl)-piperazin-1-yl]-pyrimidine-2,4,6-trione, Roche Diagnostics) and Batimastat (BB-94, Tocris Bioscience) is prepared in dimethylsulfoxide (DMSO) (Sigma) [ 54 ].

Techniques: