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 Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: H1N1 infection affects cell viability, inflammatory cytokine secretion and interactions between HBEpiCs and THP-1 cells. (A) CCK-8 assay revealed that HBEpiC viability decreased in a concentration-dependent manner following H1N1 infection. (B) ELISA revealed that the levels of IL-1β, IL-6, TNF-α, and IL-8 in HBEpiCs decreased with increasing H1N1 infection. (C) CCK-8 assay indicated that supernatants from H1N1-infected HBEpiC cultures reduced the viability of THP-1 cells in a dose-dependent manner. (D) ELISA results suggested that the levels of inflammatory cytokines (IL-1β, IL-6, TNF-α and IL-8) in THP-1 cells were decreased following exposure to supernatants from H1N1-infected HBEpiC cultures. (E) Cell adhesion assay revealed that the number of THP-1 cells adhering to HBEpiCs increased with increasing H1N1 concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) Transwell assay suggested that H1N1 infection enhanced the migration capacity of THP-1 cells, with increased migration observed at higher virus concentrations (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; ** P<0.01, *** P<0.001 vs. control. H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control; MOI, multiplicity of infection.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Infection, CCK-8 Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Adhesion Assay, Transwell Assay, Migration, Virus, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: TP modulates the inflammatory response and immune cell activity in H1N1-infected HBEpiCs and THP-1 cells. (A) No significant changes were observed in HBEpiCs treated with various concentrations of TP (5, 10 and 20 nM) following H1N1 infection compared with the control. (B) After TP treatment, the levels of the inflammatory cytokines IL-1β, IL-6, TNF-α and IL-8 in HBEpiCs were markedly lower than those in the untreated group. (C) The viability of THP-1 cells pretreated with H1N1-infected HBEpiC culture supernatant decreased after TP treatment. (D) The levels of IL-1β, IL-6, TNF-α, and IL-8 in THP-1 cells were markedly lower after TP treatment. (E) The adhesion of THP-1 cells to HBEpiCs induced by H1N1 infection decreased in a dose-dependent manner with increasing TP concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) The migration capacity of THP-1 cells was markedly reduced when the supernatant from H1N1-infected HBEpiC cultures was treated with TP (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; * P<0.05, ** P<0.01, *** P<0.001 vs. control. TP, triptolide; H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Activity Assay, Infection, Control, Concentration Assay, Migration, Standard Deviation