apelin receptor antagonist ml221  (TargetMol)

Journal: Theranostics

Article Title: Exercise ameliorates osteopenia in mice via intestinal microbial-mediated bile acid metabolism pathway

doi: 10.7150/thno.104186

Figure Lengend Snippet: FMT from exercised mice activated apelin signaling pathway. (A) PCA plot of samples from TranspCtrl and TranspExer groups (n = 3 per group). (B) Volcano plot of DEGs. (C) Enrichment analysis of upregulated DEGs in the TranspExer group. (D) Heatmap of expression levels of genes associated with the apelin signaling pathway. (E) Relative mRNA expression levels of Apln , Aplnr , Adcy4 , and Adcy5 . (F) Serum apelin concentration. (G) Representative apelin-stained IHC sections. (H) Relative mRNA expression levels of Pparg , Cebpa , Fabp4 , and Lpl . (I) Representative H&E sections depicting adipose tissue. (J-K) Quantitative analysis of J) number of adipocytes and K) adipocytes area. (L) Representative µCT images of tibias in TranspCtrl and TranspExer mice treated with AAV-Ctrl or AAV-Apln. (M-N) Trabecular bone microarchitecture showing BMD and Tb. N. Graphs show mean ± SEM (n = 6 per group), with statistical significance determined by two-tailed student t test in E-F, H, and J-K, and one-way ANOVA followed by Bonferroni's multiple comparisons test in M-N. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The apelin signaling pathway was suppressed using the apelin receptor antagonist ML221 (TargetMol, USA), wherein mice were administered ML221 (150 μg/kg) via tail vein injections three times a week for 8 weeks .

Techniques: Expressing, Concentration Assay, Staining, Two Tailed Test

Journal: Theranostics

Article Title: Exercise ameliorates osteopenia in mice via intestinal microbial-mediated bile acid metabolism pathway

doi: 10.7150/thno.104186

Figure Lengend Snippet: Exercise-matched FMT reshaped the gut metabolite profiles in OVX mice. (A) PCA plot. (B) Volcano plot of differential metabolites. (C) Enrichment analysis of upregulated metabolites in the TranspExer group. (D) Heatmap of differential metabolites between the TranspCtrl and TranspExer groups. (E) Flowchart of the gavage procedure for key metabolites in OVX mice. (F) Representative µCT images of tibias from mice in Sham, Ctrl, Tau, and UDCA groups. (G) Representative H&E-stained, OCN-stained, and TRAP-stained sections. Black arrows indicating osteoclasts. (H-I) Quantitative results of N. OBs/BS and N. OCs/BS. (J) Relative RNA expression levels of genes Apln and Aplnr . Graphs show mean ± SEM (n = 6 per group), with statistical significance determined by one-way ANOVA followed by Bonferroni's multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The apelin signaling pathway was suppressed using the apelin receptor antagonist ML221 (TargetMol, USA), wherein mice were administered ML221 (150 μg/kg) via tail vein injections three times a week for 8 weeks .

Techniques: Staining, RNA Expression

Journal: Theranostics

Article Title: Exercise ameliorates osteopenia in mice via intestinal microbial-mediated bile acid metabolism pathway

doi: 10.7150/thno.104186

Figure Lengend Snippet: Exercise-matched FMT and key metabolites ameliorated age-related bone loss. (A) Flowchart of FMT administration in aged mice. (B) Representative µCT images of tibias from aged mice in the TranspCtrl group and the TranspExer group. (C-E) Trabecular bone microarchitecture showing C) BMD, D) BV/TV, and E) Tb. N. (F) Ct. Th statistical results. (G-H) Biomechanical results of G) yield point and H) ultimate force. (I) Representative H&E-stained, OCN-stained, and TRAP-stained sections from the TranspCtrl group and the TranspExer group. Black arrows indicating osteoclasts. (J) Quantitative results of N. OBs/BS. (K) Relative mRNA expression levels of genes Apln and Aplnr . (L) Flowchart of metabolites supplementation in aged mice. (M) Representative µCT images of tibias from aged mice in the AgeCtrl, AgeTau, and AgeUDCA groups. (N-P) Trabecular bone microarchitecture showing N) BMD, O) BV/TV, and P) Tb. N. (Q) Representative H&E-stained, OCN-stained, and TRAP-stained sections from the AgeCtrl, AgeTau, and AgeUDCA groups. Black arrows indicating osteoclasts. (R) Quantitative results of N. OBs/BS. (S) Relative mRNA expression levels of genes Apln and Aplnr . Graphs show mean ± SEM (n = 6 per group), with statistical significance determined by two-tailed student t test in C-H and J-K, and one-way ANOVA followed by Bonferroni's multiple comparisons test in N-S. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The apelin signaling pathway was suppressed using the apelin receptor antagonist ML221 (TargetMol, USA), wherein mice were administered ML221 (150 μg/kg) via tail vein injections three times a week for 8 weeks .

Techniques: Staining, Expressing, Two Tailed Test

Journal: Journal of molecular and cellular cardiology

Article Title: MiR-185-5p regulates the development of myocardial fibrosis.

doi: 10.1016/j.yjmcc.2021.12.011

Figure Lengend Snippet: Fig. 4. MiR-185-5p targets apelin receptor. Human cardiac fibroblasts (hCFs) were treated with mimic-185-5p (n = 3) or mimic-Ctrl (n = 2) for 24 h and samples were subjected to RNA sequencing. Shown are (A) Gene Ontology (GO) enrichment analysis and (B) Reactome pathway analysis for the genes regulated by mimic-185-5p. (C) Schematic presentation of the binding sequence of miR-185-5p and the 5′ untranslated region (UTR) of human apelin receptor (APLNR). (D) Human umbilical vein endothelial cells were treated with mimic-185-5p (50 nM) or mimic-Ctrl. Shown is qPCR analysis for APLNR. (E-F) hCFs were treated with mimic-185-5p (20 nM) or 50 nM antimiR-185-5p and with respective controls (mimic-Ctrl or antimiR-Ctrl) for 36 h. Shown are Western blot and densitometry analyses for expression of APLNR. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. N = 3. (G) hCFs were treated with 20 nM mimic-185-5p for 24 h, and then co-transfected with mutated (mutAPLNR) or wild-type (wtAPLNR) APLNR luciferase reporter construct. Shown is normalized firefly luciferase activity (FLuc) to Renilla luciferase (RLuc) activity. N = 6. N represents the number of biological replicates. Data are shown as mean ± SD. Student's t-test was used for statistical comparison, and significance was defined as *p < 0.05, **p < 0.01 vs Ctrl.

Article Snippet: Recombinant human transforming growth factor β1 (TGF-β1, Peprotech, #100–21) was used at 5 ng/mL, apelin-13 (Phoenix Peptides, #057–18) at 200 nM and apelin receptor antagonist ML221 (Selleckchem/Absource, S8695) at 5 μM and 10 μM concentrations.

Techniques: RNA Sequencing, Binding Assay, Sequencing, Western Blot, Expressing, Control, Transfection, Luciferase, Construct, Activity Assay, Comparison

Journal: Journal of molecular and cellular cardiology

Article Title: MiR-185-5p regulates the development of myocardial fibrosis.

doi: 10.1016/j.yjmcc.2021.12.011

Figure Lengend Snippet: Fig. 5. MiR-185-5p inhibits the anti-fibrotic effects of apelin. (A) Human cardiac fibroblasts (hCFs) were treated with mimic-185-5p (N = 3) or mimic-Ctrl (N = 2) for 24 h and RNA sam ples were subjected to RNA sequencing. Shown is heatmap presentation of expression of extracellular matrix and transforming growth factor β -related components that were significantly altered by mimic-185-5p. (B) hCFs were treated with TGF-β1 (5 ng/ mL) and with mimic-185-5p (20 nM) where indicated. Shown is analysis for collagen deposition. N = 5–6. (C) TGF-β1 treated hCFs were co-treated with apelin-13 (200 nM) with or without mimic-185-5p (20 nM). Shown is analysis for collagen deposition (left panel, N = 5–6) and qPCR analysis for expression of collagen I (COL1A1, right panel, N = 4). (D) hCFs were treated apelin receptor antagonist ML221 (5 μM or 10 μM) and co-treated with antimiR-185-5p (50 nM) where indicated. Shown is analysis for collagen deposition and cell metabolic ac tivity (N = 5–6). N represents the number of biological replicates. Data are expressed as mean ± SD. One-way ANOVA followed by Tukey's post hoc test and Student's t-test was used for statistical comparison. Statistical significance was defined as **p < 0.01, ***p < 0.001 vs Ctrl; #p < 0.05, ##p < 0.01 vs TGF-β1; ††p < 0.01 vs TGF-β1 + apln-13; $p < 0.05, $$p < 0.01, $$$p < 0.001 vs anti miR-185-5p.

Article Snippet: Recombinant human transforming growth factor β1 (TGF-β1, Peprotech, #100–21) was used at 5 ng/mL, apelin-13 (Phoenix Peptides, #057–18) at 200 nM and apelin receptor antagonist ML221 (Selleckchem/Absource, S8695) at 5 μM and 10 μM concentrations.

Techniques: RNA Sequencing, Expressing, Comparison