Journal: Acta Physiologica (Oxford, England)

Article Title: Hypotaurine Reduces Glucose‐Mediated Vascular Calcification

doi: 10.1111/apha.70075

Figure Lengend Snippet: Glucose promoted extracellular matrix (ECM) calcification in primary human coronary artery smooth muscle cells (pSMC) concentration‐ and time‐dependently. pSMC were cultured in control (CM; 5.5 mM glucose) or calcium/phosphate (CaP)‐enriched media with 0, 5.5, or 25 mM glucose over 7 days. Mannitol served as osmotic control. (A) Representative images of ECM mineral accessed by Alizarin Red staining after treatment with glucose and mannitol in CM or CaP media for 1, 3, 5, and 7 days. Scale bars: 1000 μm. (B) Quantification of eluted Alizarin Red staining from the ECM from Figure . Two‐way ANOVA with Dunnett's post hoc test. * p < 0.05 compared to all other conditions at day 7. (C) Representative images of cell viability visualized with fluorescein diacetate (FDA) and propidium iodide (PI) dual immunofluorescence staining at day 7. Negative control: Permeabilized cells with 0.05% Triton X‐100. Phase contrast shows the mineral indicated by the arrow. Scale bars: 75 μm. (D) Cell viability assessed by AlamarBlue assay. pSMC were cultured with glucose and CaP media for 7 days. (E) Apoptosis at day 7. (F) Representative images of the mitochondria‐specific dye MitoTracker Red (red) and nuclear Hoechst staining (blue) at day 7. Scale bars: 75 μm. n = 3–4 in duplicates, each n represents an independent pSMC donor. Mean ± SD. One‐way ANOVA with Dunnett's post hoc test compared to 5.5 mM glucose control (CM), n.s.; not significant.

Article Snippet: Human primary coronary artery smooth muscle cells (pSMC, Promocell) were cultured in Smooth Muscle Cell Growth Medium 2 (Promocell) and supplemented with Smooth Muscle Cell Growth Medium 2 Supplement Mix (Promocell), consisting of epidermal growth factor (0.5 ng/mL), insulin (5 μg/mL), basic fibroblast growth factor‐B (2 ng/mL), 5% fetal bovine serum (FBS), and 1% penicillin–streptomycin (P/S).

Techniques: Concentration Assay, Cell Culture, Control, Staining, Immunofluorescence, Negative Control, Alamar Blue Assay

Journal: Acta Physiologica (Oxford, England)

Article Title: Hypotaurine Reduces Glucose‐Mediated Vascular Calcification

doi: 10.1111/apha.70075

Figure Lengend Snippet: Distinct glucose concentration‐specific gene expression profiles in primary human coronary artery smooth muscle cells. (A) Principal component analysis plot showed the segregation of the genes according to control (CM; 5.5 mM glucose), calcium/phosphate (CaP), and glucose treatment. The percentages indicate the proportion of variance explained by each feature. (B) The Venn diagram displayed the shared and unique differentially expressed genes (fold change ±1.2, p < 0.05) between different glucose treatments in CaP. n = 3, each n represents an independent pSMC donor.

Article Snippet: Human primary coronary artery smooth muscle cells (pSMC, Promocell) were cultured in Smooth Muscle Cell Growth Medium 2 (Promocell) and supplemented with Smooth Muscle Cell Growth Medium 2 Supplement Mix (Promocell), consisting of epidermal growth factor (0.5 ng/mL), insulin (5 μg/mL), basic fibroblast growth factor‐B (2 ng/mL), 5% fetal bovine serum (FBS), and 1% penicillin–streptomycin (P/S).

Techniques: Concentration Assay, Gene Expression, Control

Journal: Acta Physiologica (Oxford, England)

Article Title: Hypotaurine Reduces Glucose‐Mediated Vascular Calcification

doi: 10.1111/apha.70075

Figure Lengend Snippet: Glucose concentrations and time points shape unique metabolomic profiles in calcifying primary human coronary artery smooth muscle cells (pSMC). (A) Venn diagram displayed the shared and unique metabolites between supernatant and cells (calcifying SMCs). The principal component analysis (PCA) plot showed the segregation of the metabolites according to glucose treatment in (B) supernatant and (C) cells. PCA analysis was performed to distinguish between calcifying supernatant and cell samples across different timepoints. PCA biplots illustrate the positions of metabolites, with vectors indicating their relative contributions to the principal components. (D–F) Supernatant at day 3. (G–I) Supernatant at day 5. (J–L) Cell metabolites at day 3. (M–O) Cell metabolites at day 5. n = 3, each n represents an independent pSMC donor. Fold change ±1.2, p < 0.05.

Article Snippet: Human primary coronary artery smooth muscle cells (pSMC, Promocell) were cultured in Smooth Muscle Cell Growth Medium 2 (Promocell) and supplemented with Smooth Muscle Cell Growth Medium 2 Supplement Mix (Promocell), consisting of epidermal growth factor (0.5 ng/mL), insulin (5 μg/mL), basic fibroblast growth factor‐B (2 ng/mL), 5% fetal bovine serum (FBS), and 1% penicillin–streptomycin (P/S).

Techniques:

Journal: Acta Physiologica (Oxford, England)

Article Title: Hypotaurine Reduces Glucose‐Mediated Vascular Calcification

doi: 10.1111/apha.70075

Figure Lengend Snippet: The multi‐omics network of hyperglycemia‐induced vascular calcification. (A) Multi‐omics network based on genes and metabolites differentially regulated, comparing 0 vs. 25 mM glucose treatment of calcifying human coronary artery smooth muscle cells. (B) Focused network on hypotaurine/taurine and cysteine metabolic pathways and their interactors. Blue nodes represent input molecules. Yellow nodes represent metabolites, and red nodes are based on protein–protein interactions from genes. Input: Differentially expressed genes (fold change 1.5, p < 0.05, day 3) and metabolites from the supernatant (fold change 1.2, p < 0.05, day 5) from the 0 vs. 25 mM glucose comparison. (C) Overview of the effect of glucose on the hypotaurine/taurine metabolic pathway. Blue: Decreased metabolites by glucose. Orange: Increased metabolites by glucose. White: Not regulated. Green: Enzyme gene names. (D) Abundance of secreted/extracellular hypotaurine based on an untargeted metabolomics approach. n = 3, each n represents an independent pSMC donor. Mean ± SD. One‐way ANOVA with Tukey's post hoc test.

Article Snippet: Human primary coronary artery smooth muscle cells (pSMC, Promocell) were cultured in Smooth Muscle Cell Growth Medium 2 (Promocell) and supplemented with Smooth Muscle Cell Growth Medium 2 Supplement Mix (Promocell), consisting of epidermal growth factor (0.5 ng/mL), insulin (5 μg/mL), basic fibroblast growth factor‐B (2 ng/mL), 5% fetal bovine serum (FBS), and 1% penicillin–streptomycin (P/S).

Techniques: Biomarker Discovery, Protein-Protein interactions, Comparison