sacubitrilat lbq657  (MedChemExpress)

Journal: Advanced Science

Article Title: Consequences of Amyloid‐β Deficiency for the Liver

doi: 10.1002/advs.202307734

Figure Lengend Snippet: Inhibition of neprilysin protects BL/6 mice from CCl 4 ‐induced fibrosis. A) Schematic presentation of treatment timeline: 5‐week CCl 4 ‐ treatment (2x/week) of BL/6 mice ± two different dosages of neprilysin inhibitor sacubitrilat (LBQ657), 5 mg or 30 mg kg −1 body weight versus corn oil‐treated BL/6 controls (BL/6‐Ctrl.); B) plasma liver enzymes (AST, ALT, and AP), n = 8 per group; C) Sirius Red staining ( n = 4 per group); D) Immunofluorescence staining of liver sections for Col1/ αSMA/DAPI, ( n = 4 per group); E) Western Blot of αSMA and CD31 ( n = 3 per group); F–I) Multiplex analysis of Osteopontin (OPN), TGFβ, TNFα, IL‐6, IL‐13, VEGF‐A, IFNγ, MMP‐12, and MMP‐9 in liver homogenates ( n = 8 per group); The data are presented as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.001; One‐way ANOVA with Bonferroni‘s post hoc test (B, F–I) and Kruskal–Wallis test for AP analysis (in B).

Article Snippet: Two groups of BL/6 mice ( n = 8) received either 5 or 30 mg kg −1 body weight sacubitrilat (LBQ657, Hoelzel Diagnostika GmBH, Germany), injected daily over the entire period of 5‐week CCl 4 treatment.

Techniques: Inhibition, Clinical Proteomics, Staining, Immunofluorescence, Western Blot, Multiplex Assay

Journal: Advanced Science

Article Title: Consequences of Amyloid‐β Deficiency for the Liver

doi: 10.1002/advs.202307734

Figure Lengend Snippet: Schematic presentation of processes regulated by Aβ in the liver. APP‐ and NOTCH‐cleaving enzyme PSEN1 favors NOTCH processing in the situation of decreased hepatic levels of Aβ during fibrosis. Left panel: neutralization of Aβ by 3D6‐antibody treatment (anti‐Aβ) or induction of liver fibrosis in wild type (WT) mice by CCl 4 or amyloid precursor protein (APP) knockout (APP‐KO) results in downregulation of Aβ in the liver leading to decreased ammonia detoxification by glutamine synthetase (GS), HSC activation, extracellular matrix (ECM) deposition, decreased liver endothelial cell (LSEC) permeability reflected by downregulation of eNOS and VEGF and an increased expression of CD31 in transgenic 5xFAD, CCl 4 ‐treated wild type (WT) and APP‐KO mice; knockdown of APP in human liver spheroids induces epithelial‐mesenchymal transition (EMT) of hepatocytes. Right panel: high systemic and hepatic levels of Aβ in 3xTg‐AD mice and treatment of BL/6 mice with the neprilysin inhibitor sacubitrilat provide protection against liver fibrosis, normalizing the processes presented in the right panel. High hepatic Aβ level is accompanied by an overweight of PSEN1‐mediated APP cleavage over its NOTCH‐cleaving function. The figure was created with BioRender.com.

Article Snippet: Two groups of BL/6 mice ( n = 8) received either 5 or 30 mg kg −1 body weight sacubitrilat (LBQ657, Hoelzel Diagnostika GmBH, Germany), injected daily over the entire period of 5‐week CCl 4 treatment.

Techniques: Neutralization, Knock-Out, Activation Assay, Permeability, Expressing, Transgenic Assay, Knockdown

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Angiotensin Receptor–Neprilysin Inhibition (Sacubitril/Valsartan) Reduces Structural Arterial Stiffness in Middle‐Aged Mice

doi: 10.1161/JAHA.123.032641

Figure Lengend Snippet: A , Soluble collagen content in supernatant from hAoSMCs treated with TGF‐β±BNP or untreated (ctrl). Data from n=5 independent experiments. Values are individual data points, presented as mean±SEM. B , Soluble collagen content in supernatant from TGF‐β+BNP‐stimulated hAoSMCs cotreated with Val, neprilysin inhibitor LBQ657 (LBQ; active metabolite of sacubitril), valsartan+LBQ657 (Val+LBQ), or untreated (ctrl). Data from n=5 independent experiments. Values are individual data points, presented as mean±SEM. hAoSMCs from corresponding experiments were stained for collagen I (red) ( C ), and mean fluorescence (normalized to cell number) was quantified in 3 high‐power fields of 5 independent experiments. Values are individual data points, presented as mean±SEM ( D ). Nuclei are Hoechst stained (blue), and scale bar indicates 100 μm ( C ). ARNI indicates angiotensin receptor–neprilysin inhibitor; BNP, brain natriuretic peptide; ctrl, control; hAoSMCs, human aortic smooth muscle cells; n.s., not significant; TGF‐β, transforming growth factor‐β; and Val, valsartan.

Article Snippet: BNP (10 nM; TOCRIS Bioscience, Bristol, UK), valsartan (1 μM; provided by Novartis), and LBQ657 (10 μM; provided by Novartis) were added as indicated 1 hour before stimulation.

Techniques: Staining, Fluorescence

Journal: Frontiers in Cardiovascular Medicine

Article Title: Sacubitril/valsartan attenuates atrial conduction disturbance and electrophysiological heterogeneity with ameliorating fibrosis in mice

doi: 10.3389/fcvm.2024.1341601

Figure Lengend Snippet: BNP and CNP suppressed the activation of primary adult cardiac fibroblasts. ( A ) The protocol of the in vitro assay. ( B ) Effects of ANP (left), BNP (middle) or CNP (right) (at 1.0 µ M each) against the pro-fibrotic alteration of Ccn2 and Acta2 induced by TGFβ ( n = 5–8). ( C ) Additional effect of LBQ657 (50 µ M) on BNP (left) or CNP (right) (5.0 µ M each) ( n = 6–7). ( D ) Antagonizing actions of P19 (1.0 µ M) on CNP (1.0 µ M) added with LBQ657 (10 µ M) ( n = 6). ( E ) Effects of AP811 (1.0 µ M) on CNP (1.0 µ M) with LBQ657 (10 µ M) ( n = 7). Multiple comparison was performed using one-way ANOVA with Tuckey's post-hoc test. Error bars, SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. ANP, atrial natriuretic peptide; BNP, B-type natriuretic peptide; CNP, C-type natriuretic peptide; TGFβ, transforming growth factor β.

Article Snippet: At the same time, a pre-treatment with Atrial Natriuretic Peptide (ANP) (Phoenix Pharmaceuticals, Inc., Burlingame, California, USA), B-type Natriuretic Peptide (BNP) (Bachem, Bubendorf, Switzerland) or C-type Natriuretic Peptide (CNP) (MedChemExpress, Monmouth Junction, New Jersey, USA) at 1.0 or 5.0 µ M was initiated with or without antagonists of their cognate receptors; A71915 (Sigma Aldrich, Burlington, Vermont, USA) antagonizing natriuretic peptide receptor-A (NPR-A); P19 (Phoenix Pharmaceuticals, Inc.) antagonizing natriuretic peptide receptor-B (NPR-B); AP811 (R&D Systems, Minneapolis, Minnesota, USA) antagonizing natriuretic peptide receptor-C (NPR-C), at 1.0 µ M each, as well as LBQ657 (sacubitrilat; Cayman Chemical, Ann Arbor, Michigan, USA) at 10 or 50 µ M. Transforming growth factor β (TGFβ; PeproTech, Cranbury, New Jersey, USA) (1.0 ng/ml) was added 30 min after the application of NPs, followed by another incubation for 24 h. After incubation with the drugs, RNA was extracted from the cultured cells and reverse transcription was performed as described above.

Techniques: Activation Assay, In Vitro, Comparison