Journal: Aging Cell

Article Title: Telomerase Knockout in Myeloid Cells Predisposes Mice to Foam Cell Formation, Dyslipidemia, Lung Fibrosis, and Cardiac Dysfunction

doi: 10.1111/acel.70490

Figure Lengend Snippet: Tert KO induces lipid‐associated macrophages (LAMs). Starch‐induced i.p. macrophages from 2‐year‐old female mice were analyzed. (a) Upon adherence in primary culture, IF with antibodies against CD80 (M1‐macrophage) and CD206 (M2‐polarization) markers reveals a lower frequency of CD206 + macrophages in KO mice. (b) Data quantification from multiple fields of view in (a), indicating macrophage polarization shift toward the M1 phenotype. (c) Upon LPS (100 ng/mL, 4 h) treatment in primary culture, q‐RT‐PCR (normalized to 18S RNA) demonstrates higher expression of genes coding for inflammation markers IL1 and IL6 in KO macrophages. (d) Macrophages were induced to convert into foam cells by oxLDL (0.025 mg/mL) treatment for 24 h. Note increased uptake of red‐fluorescent C 12 ‐BODIPY (0.3 μM, 5 min) by mG+ KO cells (yellow arrows) compared to mG+ WT cells (green arrows) in primary culture. (e) q‐RT‐PCR (normalized to 18S RNA) demonstrates lower expression of genes coding for lipid efflux effectors APOE, LDLR, ABCA1, ABCG1, and higher expression of genes coding for lipid transporters CD36 and FABP5 in KO oxLDL‐treated macrophages. (f) OxLDL‐treated macrophages stained with Oil Red O: Note larger lipid droplets (arrows) in KO cells. For all data, mean+/− SEM (error bars). * p < 0.05, ** p < 0.01, *** p < 0.001 (two‐sided Student's t ‐test). Scale bar: 50 μm.

Article Snippet: Prior to Oil Red O staining (Sigma‐Aldrich, O0625) and BODIPY‐FL‐C 12 (Invitrogen, D3822) uptake analysis, oxLDL (Athensresearch, 12‐16‐120412‐OX) was added to the medium at 25 μg/mL for 24 h.

Techniques: Starch, Reverse Transcription Polymerase Chain Reaction, Expressing, Staining

Journal: Frontiers in Cardiovascular Medicine

Article Title: Establishing an atherosclerosis diagnostic model based on WGCNA and machine learning algorithms with key genes in cholesterol metabolism and ferroptosis, and revealing the regulatory role of HMOX1 in cellular ferroptosis

doi: 10.3389/fcvm.2026.1715946

Figure Lengend Snippet: HMOX1 promotes ox-LDL-induced dysfunction of foam macrophages by regulating ferroptosis. (A) Expression of diagnostic genes in normal and AS groups. (B) Expression of diagnostic genes in AS progression and regression groups. The progression group and regression group were isolated from Cx3cr1 CreERT2−IRES−YFP/+ ; Rosa26 floxed−tdTomato/+ mice treated with AAV-mPCSK9 and subjected to dietary intervention. (C) Single-cell RNA sequencing data of the GSE155512 dataset, processed by the “Seurat” package and subjected to UMAP dimensionality reduction, showing the main cell type composition in the atherosclerotic samples. (D) Expression distribution of HMOX1 in different cell types. (E,F) Expression of HMOX1 in ox-LDL-induced THP-1 derived foam macrophages and verification of knockdown by qRT-PCR (E) and Western blot (F) (G) Effects of HMOX1 knockdown on macrophage viability under different conditions assessed by CCK-8 assay. (H–J) Effects of HMOX1 knockdown on oxidative stress (H) , lipid peroxidation (I) , and iron accumulation (J) induced by ox-LDL. (K) : Effects of HMOX1 knockdown on the expression of the key ferroptotic gene ACSL4 and anti-ferroptotic genes SLC7A11 and GPX4. All in vitro experiments were independently repeated three times. Continuous variables were expressed as mean ± standard deviation. Comparisons between two groups were performed using Student's t -test, and comparisons among multiple groups were evaluated using one-way ANOVA. Single-cell data analysis was based on R (version 4.4.2) and was mainly completed using R packages such as “Seurat” and “Harmony”. Statistical analysis and visualization were performed in R software and GraphPad Prism 10.1.2. ns: no statistical difference, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001.

Article Snippet: Differentiation into macrophages was induced by stimulating THP-1 cells with 50 nM phorbol myristate acetate (16561-29-8, Merck, USA) for 48 h. Then, 100 μg/mL oxidized low-density lipoprotein (ox-LDL) (308068-14-6, Merck, USA) was applied for 48 h to form foam-like macrophages.

Techniques: Expressing, Diagnostic Assay, Isolation, Single Cell, RNA Sequencing, Derivative Assay, Knockdown, Quantitative RT-PCR, Western Blot, CCK-8 Assay, In Vitro, Standard Deviation, Software

Journal: International Journal of Molecular Sciences

Article Title: Twenty-Four-Month rhGH Intervention: Insights into Redox Regulation, Vascular Biomarkers, and Body Composition in Adult GHD Patients

doi: 10.3390/ijms27031451

Figure Lengend Snippet: Longitudinal changes in ox-LDL, Trx, E-selectin, P-selectin, ICAM-1, and VCAM-1 during 24-month rhGH therapy at baseline, 12 months, and 24 months. Statistical significance is indicated by asterisks: * p < 0.05 vs. baseline; *** p < 0.001 vs. baseline; **** p < 0.00001 vs. baseline.

Article Snippet: OS and endothelial markers were quantified using commercially available enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturers’ instructions: ox-LDL (Novus Biologicals, Bio-Techne, NBP3-18794; Minneapolis, MN, USA), Trx (XpressBio, XPEH0291; Frederick, MD, USA), OGG1 (Cloud-Clone Corp., SEC704Hu; Wuhan, China), VCAM-1 (Cloud-Clone Corp., SEA547Hu; Wuhan, China), ICAM-1 (Cloud-Clone Corp., SEA548Ca; Wuhan, China), and E-selectin (Cloud-Clone Corp., SEA029Hu; Wuhan, China).

Techniques:

Journal: European Journal of Medical Research

Article Title: lncRNA SND1-IT1/miR-494-3p regulation in ox-LDL-induced endothelial cell injury: novel insights into coronary heart disease pathogenesis and diagnostic biomarkers

doi: 10.1186/s40001-026-03918-8

Figure Lengend Snippet: ox-LDL effects on HUVECs. a – b Expression levels of SND1-IT1 ( a ) and miR-494-3p ( b ) measured by RT-qPCR after treatment with increasing concentrations of ox-LDL. c Cell viability assessed by CCK-8 assay (72 h). d Apoptosis rate determined by flow cytometry. e mRNA expression of apoptosis-related genes (Bax, Bcl-2) analyzed by RT-qPCR. f mRNA expression of cell adhesion molecules (VCAM-1, ICAM-1) measured by RT-qPCR. g Inflammatory cytokine levels (IL-6, TNF-α) detected by ELISA. h Antioxidant enzyme activities (CAT, SOD) measured using specific kits

Article Snippet: Simultaneously, ox-LDL (sourced from Nanjing Jiancheng Bioengineering Institute, China) was diluted with PBS buffer to concentrations of 0, 25, 50, and 100 μg/mL.

Techniques: Expressing, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: European Journal of Medical Research

Article Title: lncRNA SND1-IT1/miR-494-3p regulation in ox-LDL-induced endothelial cell injury: novel insights into coronary heart disease pathogenesis and diagnostic biomarkers

doi: 10.1186/s40001-026-03918-8

Figure Lengend Snippet: The role of SND1-IT1 downregulation on ox-LDL-treated HUVECs. a SND1-IT1 expression after transfection with si-SND1-IT1 measured by RT-qPCR. b Cell viability by CCK-8 assay. c Apoptosis rate determined by flow cytometry. d mRNA expression of apoptosis-related genes (Bax, Bcl-2) analyzed by RT-qPCR. e mRNA expression of cell adhesion molecules (VCAM-1, ICAM-1) measured by RT-qPCR. f Inflammatory cytokine levels (IL-6, TNF-α) detected by ELISA. g Antioxidant enzyme activities (CAT, SOD) measured using specific kits

Article Snippet: Simultaneously, ox-LDL (sourced from Nanjing Jiancheng Bioengineering Institute, China) was diluted with PBS buffer to concentrations of 0, 25, 50, and 100 μg/mL.

Techniques: Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: European Journal of Medical Research

Article Title: lncRNA SND1-IT1/miR-494-3p regulation in ox-LDL-induced endothelial cell injury: novel insights into coronary heart disease pathogenesis and diagnostic biomarkers

doi: 10.1186/s40001-026-03918-8

Figure Lengend Snippet: SND1-IT1-miR-494-3p interaction. a Dual-luciferase reporter assay confirming direct binding. b RNA immunoprecipitation (RIP) showing enrichment of both molecules. c SND1-IT1 expression after miR-494-3p inhibition measured by RT-qPCR. d miR-494-3p expression after SND1-IT1 knockdown measured by RT-qPCR. e – j Effects of miR-494-3p downregulation on cell viability ( e , CCK-8), apoptosis ( f , flow cytometry), apoptosis-related gene mRNA ( g , RT-qPCR), cell adhesion molecule mRNA expression ( h , RT-qPCR), inflammatory cytokines ( i , ELISA), and antioxidant enzymes ( j , specific kits). ox-LDL + si-SND1: HUVECs stimulated with ox-LDL and transfected with siRNA targeting SND1-IT1; ox-LDL + si + miR-NC: ox-LDL-stimulated HUVECs co-transfected with si-SND1-IT1 and a negative control for miRNA; ox-LDL + si + miR-inhibitor: ox-LDL-stimulated HUVECs co-transfected with si-SND1-IT1 and an inhibitor of miR-494-3p

Article Snippet: Simultaneously, ox-LDL (sourced from Nanjing Jiancheng Bioengineering Institute, China) was diluted with PBS buffer to concentrations of 0, 25, 50, and 100 μg/mL.

Techniques: Luciferase, Reporter Assay, Binding Assay, RNA Immunoprecipitation, Expressing, Inhibition, Quantitative RT-PCR, Knockdown, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Transfection, Negative Control

Journal: European Journal of Medical Research

Article Title: lncRNA SND1-IT1/miR-494-3p regulation in ox-LDL-induced endothelial cell injury: novel insights into coronary heart disease pathogenesis and diagnostic biomarkers

doi: 10.1186/s40001-026-03918-8

Figure Lengend Snippet: The role of ZBTB20 overexpression in ox-LDL-treated HUVECs. a ZBTB20 expression after transfection measured by RT-qPCR. b ZBTB20 protein expression detected by Western blot. c Cell viability assessed by CCK-8 assay. d Apoptosis rate determined by flow cytometry. e mRNA expression of apoptosis-related genes (Bax, Bcl-2) analyzed by RT-qPCR. f mRNA expression of cell adhesion molecules (VCAM-1, ICAM-1) measured by RT-qPCR. g Inflammatory cytokine levels (IL-6, TNF-α) detected by ELISA. h Antioxidant enzyme activities (CAT, SOD) measured using specific kits. ox-LDL + miR-NC: HUVECs stimulated with ox-LDL and transfected with a miRNA negative control; ox-LDL + miR-mimic: ox-LDL-stimulated HUVECs transfected with a miR-494-3p mimic; ox-LDL + miR + pcDNA: ox-LDL-stimulated HUVECs co-transfected with miR-494-3p mimic and an empty pcDNA vector; ox-LDL + miR + pcDNA-ZBTB20: ox-LDL-stimulated HUVECs co-transfected with miR-494-3p mimic and a ZBTB20-overexpressing plasmid (pcDNA-ZBTB20)

Article Snippet: Simultaneously, ox-LDL (sourced from Nanjing Jiancheng Bioengineering Institute, China) was diluted with PBS buffer to concentrations of 0, 25, 50, and 100 μg/mL.

Techniques: Over Expression, Expressing, Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Negative Control, Plasmid Preparation