Journal: Frontiers in Pharmacology

Article Title: Gastrodin mitigates aortic valve calcification by inhibiting glycolysis and histone lactylation through interfering with valve interstitial cells

doi: 10.3389/fphar.2025.1547716

Figure Lengend Snippet: GAS inhibits the expression of osteogenesis-specific markers induced by OM. (A) RT-qPCR results show that GAS can significantly inhibit the expression of specific markers of osteogenesis in OM-induced VICs (OMI, OM-induced VICs group; GAS 50 μM treatment group; GAS 100 μM treatment group; AGP 10 μM treatment group) (n = 3). (B) Western blot results show that GAS can significantly inhibit the expression of specific markers of osteogenesis in OM-induced VICs (n = 3); (C) Semi-quantitative statistics of the band thickness of the Western blot results in B; (D) In-cell Western blot results show GAS can significantly inhibit the expression of osteogenic specific markers in OM-induced VICs (n = 3); (E) Semi-quantitative statistics of the intensity of In-cell blotting results in (D) Statistical significance is indicated by # for OMI group vs control group and * for GAS or AGP group vs OMI group ( p ≤ 0.05).

Article Snippet: Osteogenic medium (OM, Stem Cell Technologies Cat No. 05465) was continuously treated for 12 days to induce osteogenic differentiation of VICs.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, In-Cell ELISA, Control

Journal: Frontiers in Pharmacology

Article Title: Gastrodin mitigates aortic valve calcification by inhibiting glycolysis and histone lactylation through interfering with valve interstitial cells

doi: 10.3389/fphar.2025.1547716

Figure Lengend Snippet: Gene expression profiles of mRNA sequencing with gastrodin (GAS) treatment under the condition of osteogenic medium induction (OMI). (A,B) Volcano map of differentially expressed genes (DEGs) in OMI versus control (OMI/Con), upregulation: 823 and downregulation: 484; in GAS versus OMI (GAS/OMI), upregulation: 247 and downregulation: 305, |log2 (fold change) (FC)| > 1 and false discovery rate (FDR) < 0.001 was considered as DEGs; (C) Venn interaction of DEGs of OMI/Con and GAS/OMI; (D) GO enrichment of the common DEGs including biological process, cellular component and molecular function; (E) KEGG pathway enrichment; a lower Q value indicates a higher degree of enrichment.

Article Snippet: Osteogenic medium (OM, Stem Cell Technologies Cat No. 05465) was continuously treated for 12 days to induce osteogenic differentiation of VICs.

Techniques: Gene Expression, Sequencing, Control

Journal: Frontiers in Pharmacology

Article Title: Gastrodin mitigates aortic valve calcification by inhibiting glycolysis and histone lactylation through interfering with valve interstitial cells

doi: 10.3389/fphar.2025.1547716

Figure Lengend Snippet: Metabolome analysis and expression evaluation of osteogenic medium-induced and gastrodin-treated VICs. (A) Total ion flow diagram of cell metabolites (n = 10); (B) Principal component analysis (PCA) diagram, (C,D) orthogonal partial least squares discrimination analysis diagram (OPLS-DA) (n = 10); (E) Cluster analysis of metabolome of OM-induced and GAS-treated VICs (n = 10); (F) Pathway enrichment based on differential metabolites (VIP>1), glycolysis/gluconeogenesis, pyruvate metabolism, fructose/mannose metabolism and glycerolipid metabolism are highlighted; (G) Top differential metabolite analysis including acetic acid and lactic acid, * p < 0.05 versus OMI (n = 10); (H) Glycolytic key gene expression detection of VICs with OM and OM plus GAS treatment (n = 3). # p < 0.05 versus control (con), * p < 0.05 versus OM.

Article Snippet: Osteogenic medium (OM, Stem Cell Technologies Cat No. 05465) was continuously treated for 12 days to induce osteogenic differentiation of VICs.

Techniques: Expressing, Gene Expression, Control

Journal: Frontiers in Pharmacology

Article Title: Gastrodin mitigates aortic valve calcification by inhibiting glycolysis and histone lactylation through interfering with valve interstitial cells

doi: 10.3389/fphar.2025.1547716

Figure Lengend Snippet: Gastrodin inhibits VICs calcification via H3 histone lactylation by interfering with PKM2. (A) Pan Kla expression levels of VICs with osteogenic medium (OM) and OM plus gastrodin (GAS) treatments (n = 3); (B) The binding energy of gastrodin and corelating proteins; (C) Molecular docking of gastrodin and the proteins with 4 least binding energy PKM2, ALDOA, ALDOB, LDHB; (D) Enzyme activity of PKM2 in VICs treated with gastrodin and untreated with gastrodin protein under OM induction (n = 3); (E,F) Expression levels of PKM2, SPP1, COL1A2, RUNX2, BMP2 and H3 Kla of VICs under OM inducing with gastrodin and siPKM2 treating (n = 3); (G) Enzyme activity of PKM2 in VICs under OM inducing with gastrodin and siPKM2 treating (n = 3); (H) Alizarin Red staining in VICs under OM inducing with gastrodin and siPKM2 treating (n = 3); (I,J) Expression levels of BMP2, PKM2 and H3 Kla of VICs under OM inducing with and without gastrodin treating (n = 3); (K) Correlation analysis of the expression data above; ( L,M) In-cell Western blotting of BMP2, RUNX2, PKM2, LDHA, H3K9la of VICs under OM inducing with and without gastrodin treating, Histone H3 (n = 3); (N) Correlation analysis of the expression data above. # p ≤ 0.05 in OMI vs Control, * p ≤ 0.05 in GAS-treated vs OMI.

Article Snippet: Osteogenic medium (OM, Stem Cell Technologies Cat No. 05465) was continuously treated for 12 days to induce osteogenic differentiation of VICs.

Techniques: Expressing, Binding Assay, Activity Assay, Staining, In-Cell ELISA, Control

Journal: Stem Cells International

Article Title: Differentiation of Human Embryonic Stem Cells into Neuron, Cholinergic, and Glial Cells

doi: 10.1155/2020/8827874

Figure Lengend Snippet: Flow chart of sequential procedure used to produce cholinergic cells derived from human neural stem cells in adherent culture. The protocol involves daily medium change with increasing concentration of NGF and decreasing concentration of EGF, bFGF, and B-27. The medium on day 10 was kept constant for the next four days. The protocol is previously described by .

Article Snippet: DMEM/F12 (Thermo Fisher Scientific, Cat. No 11320033) medium together with increasing concentration of NGF-beta (Stem Cell Technology, Cat. No. 78092) and reducing concentration of B-27™ Supplement, EGF (Stem Cell Technology, Cat. No. 78006), and BFGF (Stem Cell Technology, Cat. No. 788003.1) was used as complete differentiation medium.

Techniques: Derivative Assay, Concentration Assay