Journal: Mechanobiology in Medicine

Article Title: Stacked human aortic endothelial cells induce atherosclerotic fatty streaks and release proinflammatory cytokines and chemokines ☆

doi: 10.1016/j.mbm.2026.100192

Figure Lengend Snippet: RNA sequencing analysis of coralthelial cells and chemokines/cytokines detection. (A) Volcano plot of DEGs. The red and green spots indicate up- and down-regulated genes, respectively. The chemokines/cytokines IL-6, MCP-1, and CXCL8, which plays a role in human cardiovascular events, are marked in blue. (B) PCA based on the transcriptome profile indicates that the first principal component (PC1) can separate coralthelial cells (stack) from HAECs (monolayer). (C) GO analysis of DEGs indicates that the significantly enriched GO terms in coralthelial cells are related to chemokine/cytokine signaling in inflammation. (D) Pathway analysis of DEGs corroborates that chemokine/cytokine signaling pathways in inflammation are significantly altered in coralthelial cells. (E–G) qRT-PCR confirms increased mRNA expression of IL-6 (E), MCP-1 (F), and CXCL8 (G) in coralthelial cells. (H–J) ELISA detection of IL-6 (H), MCP-1 (I), and CXCL8 (J) in the culture medium of coralthelial cells and that of HAECs on days 2, 5, 8, and 11. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001 vs. HAECs (monolayer).

Article Snippet: The concentrations of IL-6, MCP-1, and CXCL8 in the culture supernatants of HAECs and coralthelial cells on days 2, 5, 8, and 11 were measured using human IL-6, MCP-1, and CXCL8 ELISA Kits (#ELH-IL6, #ELH-MCP1, and #ELH-IL8; RayBio, USA), per the manufacturer's instructions.

Techniques: RNA Sequencing, Protein-Protein interactions, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Mechanobiology in Medicine

Article Title: Stacked human aortic endothelial cells induce atherosclerotic fatty streaks and release proinflammatory cytokines and chemokines ☆

doi: 10.1016/j.mbm.2026.100192

Figure Lengend Snippet: RPL23 translocates into the nucleolus and facilitates the secretion of proinflammatory cytokines, which is attenuated by SAR1B siRNA. (A) Co-staining of RPL23 and FBL (the nucleolus marker) in monolayer HAECs and coralthelial cells. (B, C) Following SAR1B knockdown, the nuclear translocation of RPL23 (B) and GM130 (C) were assessed in coralthelial cells. Line scans were performed for RPL23 and FBL. The MFIs of RPL23, FBL, GM130, and SAR1B in the nucleus and cytoplasm are presented as fold-changes relative to their levels in the cytoplasm of monolayer HAECs or NC coralthelial cells (stack). Data are presented as the mean ± SD; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. (D) The levels of IL-6, MCP-1, and CXCL8 in coralthelial cells were quantified. ∗∗∗P < 0.001 vs. NC monolayer; ##P < 0.01, ###P < 0.01 vs. NC stack.

Article Snippet: The concentrations of IL-6, MCP-1, and CXCL8 in the culture supernatants of HAECs and coralthelial cells on days 2, 5, 8, and 11 were measured using human IL-6, MCP-1, and CXCL8 ELISA Kits (#ELH-IL6, #ELH-MCP1, and #ELH-IL8; RayBio, USA), per the manufacturer's instructions.

Techniques: Staining, Marker, Knockdown, Translocation Assay

Journal: Mechanobiology in Medicine

Article Title: Stacked human aortic endothelial cells induce atherosclerotic fatty streaks and release proinflammatory cytokines and chemokines ☆

doi: 10.1016/j.mbm.2026.100192

Figure Lengend Snippet: Major differences between human aortic endothelial cells and coralthelial cells, and the mechanism of cytokine release in coralthelial cells . (A) HAECs display a typical endothelial morphology with a flattened, elongated structure, organized actin filaments, intact glycocalyx, and minimal lipid accumulation. (B) By contrast, coralthelial cells, derived from stacked HAECs, exhibit a distinct coral-like morphology with surface blebbing, smaller cell bodies and nuclei, disorganized actin filaments, degraded glycocalyx, smaller mitochondria, enhanced nucleolar stress, and prominent lipid droplet accumulation, resembling foam cells found in atherosclerotic plaques. Coralthelial cells also lack typical endothelial markers such as PECAM-1 (CD31) and FVIII, which are expressed in HAECs. Coralthelial cells show a marked increase in collagen production, helping form the extracellular matrix scaffold observed in atherosclerotic fatty streak–like structures. In addition, coralthelial cells exhibit significant upregulation of proinflammatory cytokines and chemokines, including IL-6, MCP-1 and CXCL8, promoting a proinflammatory microenvironment characteristic of atherosclerosis. This cytokine/chemokine release by coralthelial cells is likely driven by the ER–Golgi–nucleus axis and involves the nuclear translocation of RPL23, in conjunction with the Golgi apparatus and COPII vesicles (key component SAR1B). Targeting RPL23 and ER–Golgi dynamics represents a promising therapeutic strategy for modulating cytokine release in vascular diseases, including atherosclerosis. go, Golgi apparatus; ER, endoplasmic reticulum; mi, mitochondria; LD, lipid droplet; N, nucleolus; HAECs, human aortic endothelial cells; PECAM-1, platelet endothelial cell adhesion molecule-1; FVIII, factor VIII; COPII, coat protein complex II; IL-6, interleukin-6; MCP-1, monocyte chemoattractant protein-1; CXCL8, C-X-C motif chemokine ligand 8.

Article Snippet: The concentrations of IL-6, MCP-1, and CXCL8 in the culture supernatants of HAECs and coralthelial cells on days 2, 5, 8, and 11 were measured using human IL-6, MCP-1, and CXCL8 ELISA Kits (#ELH-IL6, #ELH-MCP1, and #ELH-IL8; RayBio, USA), per the manufacturer's instructions.

Techniques: Derivative Assay, Translocation Assay

Journal: International Journal of Molecular Sciences

Article Title: ASC-Derived Extracellular Vesicles Suppress Macrophage-Driven Inflammatory Amplification and Contractile Activation of Uterine Smooth Muscle Cells

doi: 10.3390/ijms27104273

Figure Lengend Snippet: ASC-derived EVs suppress LPS-induced inflammatory signaling in HUtSMCs. ( A ) HUtSMCs were pretreated with ASC-derived EVs and stimulated with LPS for the indicated time points (5–20 min). Phosphorylation of JNK and ERK1/2 was analyzed by Western blotting. ( B ) mRNA expression levels of TSG6, IL-6, IL-8, MCP-1, and MIP-2 were measured by quantitative RT-PCR at 8–24 h after LPS stimulation and normalized to GAPDH. ( C ) Secretion levels of IL-6, IL-8, and MCP-1 were quantified by ELISA. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: IL-6, IL-8, and MCP-1 levels were measured using ELISA kits, including Human IL-6 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL6-1), Human IL-8 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL8-1), and Human MCP-1 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-MCP1-1), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: ASC-Derived Extracellular Vesicles Suppress Macrophage-Driven Inflammatory Amplification and Contractile Activation of Uterine Smooth Muscle Cells

doi: 10.3390/ijms27104273

Figure Lengend Snippet: ASC-derived EVs modulate TLR4-dependent inflammatory signaling in HUtSMCs. ( A ) HUtSMCs were pretreated with ASC-EVs and/or a TLR4 inhibitor prior to LPS stimulation. Phosphorylation of JNK and ERK1/2 was analyzed by Western blotting. Representative images from three independent experiments are shown. ( B ) mRNA expression levels of IL-6, IL-8, MCP-1, TNF-α, MIP-1α, and MIP-2 were analyzed by quantitative RT-PCR and normalized to GAPDH. Data are presented as mean ± SEM from three independent experiments and are expressed relative to the untreated control. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: IL-6, IL-8, and MCP-1 levels were measured using ELISA kits, including Human IL-6 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL6-1), Human IL-8 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL8-1), and Human MCP-1 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-MCP1-1), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR, Control

Journal: International Journal of Molecular Sciences

Article Title: ASC-Derived Extracellular Vesicles Suppress Macrophage-Driven Inflammatory Amplification and Contractile Activation of Uterine Smooth Muscle Cells

doi: 10.3390/ijms27104273

Figure Lengend Snippet: ASC-derived EVs attenuate macrophage-mediated inflammatory signaling in HUtSMCs. ( A ) HUtSMCs were pretreated with ASC-derived EVs and stimulated with conditioned media (CM) derived from RAW264.7 macrophages treated with LPS (10–100 ng/mL). Cells were harvested after 10 min, and phosphorylation of JNK and ERK1/2 was analyzed by Western blotting. ( B ) mRNA expression levels of IL-6, IL-8, MCP-1, TNF-α, MIP-1α, and MIP-2 were analyzed by quantitative RT-PCR 20 h after CM stimulation. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: IL-6, IL-8, and MCP-1 levels were measured using ELISA kits, including Human IL-6 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL6-1), Human IL-8 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL8-1), and Human MCP-1 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-MCP1-1), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: ASC-Derived Extracellular Vesicles Suppress Macrophage-Driven Inflammatory Amplification and Contractile Activation of Uterine Smooth Muscle Cells

doi: 10.3390/ijms27104273

Figure Lengend Snippet: ASC-derived EVs inhibit macrophage TLR4-dependent NF-κB and MAPK activation in HUtSMCs. ( A ) RAW264.7 macrophages were stimulated with LPS in the presence or absence of the TLR4 inhibitor TAK242. mRNA expression levels of IL-1β, IL-10, TNF-α, MIP-2, and MCP-1 were analyzed by quantitative RT-PCR and normalized to β-actin. ( B ) HUtSMCs were treated with conditioned media derived from RAW264.7 macrophages under the indicated conditions, with or without ASC-EVs pretreatment. Phosphorylation of p65, JNK, and ERK1/2 was analyzed by Western blotting. ( C ) Immunofluorescence analysis of phosphorylated p65 (green) in HUtSMCs. Nuclei were stained with DAPI (blue). ( D ) Cytoplasmic and nuclear fractionation analysis of NF-κB signaling. Levels of phosphorylated p65, total p65, phosphorylated IκB, and total IκB were analyzed by Western blotting. α-tubulin and Lamin B1 were used as cytoplasmic and nuclear markers, respectively. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: IL-6, IL-8, and MCP-1 levels were measured using ELISA kits, including Human IL-6 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL6-1), Human IL-8 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL8-1), and Human MCP-1 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-MCP1-1), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Activation Assay, Expressing, Quantitative RT-PCR, Phospho-proteomics, Western Blot, Immunofluorescence, Staining, Fractionation

Journal: bioRxiv

Article Title: Middle-aged mice treated with GHK-Cu peptide administered intraperitoneally or intranasally show behavioral rescue but divergent hippocampal aging programs

doi: 10.64898/2026.04.09.717524

Figure Lengend Snippet: Positive area-staining by antibody in intranasally-treated GHK-Cu mice of both sexes for (A) Synaptophysin, (B) GFAP, (C) MCP-1, (D) PSD-95, and (E) TGF-β. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Following blocking, sections were incubated overnight at 4°C with primary antibodies against synaptophysin (1:250, Invitrogen MA5-16402), PSD95 (1:250, Abcam ab18258), phospho-SMAD2 (1:50, Invitrogen 44-244G), MCP-1 (1:800, Novus NBP1-07035), p21 (1:200, Abcam ab188224), TGF-β (1:50, Abcam ab215715), and GFAP (1:1500, Invitrogen: PA1-10019).

Techniques: Staining

Journal: bioRxiv

Article Title: Middle-aged mice treated with GHK-Cu peptide administered intraperitoneally or intranasally show behavioral rescue but divergent hippocampal aging programs

doi: 10.64898/2026.04.09.717524

Figure Lengend Snippet: Positive area-staining by antibody in intraperitoneally-treated GHK-Cu mice of both sexes for (A) TGF-β, (B) GFAP), (C) MCP-1, (D) p21, (E) Synaptophysin, (F) pSMAD-2, and (G) PSD-95. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: Following blocking, sections were incubated overnight at 4°C with primary antibodies against synaptophysin (1:250, Invitrogen MA5-16402), PSD95 (1:250, Abcam ab18258), phospho-SMAD2 (1:50, Invitrogen 44-244G), MCP-1 (1:800, Novus NBP1-07035), p21 (1:200, Abcam ab188224), TGF-β (1:50, Abcam ab215715), and GFAP (1:1500, Invitrogen: PA1-10019).

Techniques: Staining