Journal: Experimental Biology and Medicine

Article Title: Mechanisms of AGE-induced VSMC phenotypic switching and macrophage modulation in human abdominal aortic aneurysms

doi: 10.3389/ebm.2025.10527

Figure Lengend Snippet: AGEs Activate the NF-κB Pathway and NLRP3 Inflammasome via the RAGE/RhoA/ROCK Pathway. (A) The TransAM NF-κB Transcription Factor Assay detected the effect of the RAGE/RhoA/ROCK inhibitor on the nuclear translocation of c-Rel, p52, and p65; (B) The NF-κB dual luciferase reporter gene system assessed the impact of RAGE/RhoA/ROCK inhibitors on AGE-induced NF-κB signaling; (C) Western blot analysis showed that AGEs regulate the expression of NLRP3 via the RAGE/RhoA/ROCK/NF-κB signaling pathway; (D) The effects of the inhibitors on caspase-1 activation were assessed using a caspase-1 assay kit; (E) An LDH release assay showed that AGE promoted VSMC cell pyroptosis, but it could be reversed by RAGE, RhoA, and ROCK inhibitors; (F) An ELISA assay demonstrated that AGE promoted the release of the inflammatory factor IL-1β through RAGE/RhoA/ROCK. Data are presented as the mean ± SD from three independent experiments and were analyzed with a one-way ANOVA followed by a Dunnett’s multiple comparisons test. Asterisks indicate significant differences (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001) between the treatment group and the control group. Pound signs indicate a significant difference (#P < 0.05, ##P < 0.01, ###P < 0.001) between the treatment group and the AGE group.

Article Snippet: The enzymatic activity of caspase-1 was assayed using a Caspase-1 Colorimetric Assay Kit (MCE, Cat. No. HY-K2610-100T) according to the manufacturer’s protocol.

Techniques: Transcription Factor Assay, Translocation Assay, Luciferase, Western Blot, Expressing, Activation Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Control

Journal:

Article Title: High-Level Replication of Human Immunodeficiency Virus in Thymocytes Requires NF-?B Activation through Interaction with Thymic Epithelial Cells

doi:

Figure Lengend Snippet: TNF and, to a lesser extent, IL-1 are involved in the permanently induced NF-κB activity present in thymocytes during the coculture with TEC. Thymocytes were freshly isolated (lanes 1 and 2) or cultured for 45 h (lanes 3 to 11) either alone (lane 3 [Control thymocytes]) or in the presence of TEC under conditions avoiding contact (lanes 4 to 9) or allowing contact (lanes 10 and 11). During the coculture, the cells were either left untreated (lane 4 [Control coculture, 45 hours] and lane 10 [Control coculture]) or were treated at the start of the coculture with antibodies respectively raised against TNF (lane 5), IL-6 (lane 6), and IL-1β (lane 8), with the antagonist of the IL-1 receptor (lane 7), or with αTNF plus IL-1 ra plus αIL-6 (lanes 9 and 11). Whole-cell extracts from these various samples were incubated with a 32P-labeled oligonucleotide representing the HIV LTR-derived κB motif. Competition (comp [lane 1]) with a 40-fold molar excess of unlabeled oligonucleotide was used to confirm the specificity of the DNA-binding activity detected. NF-κB activity is indicated. The positions of two nonspecific (n.s 1 and n.s 2) binding activities are indicated. The positions of two nonspecific (n.s 1 and n.s 2) binding activities are indicated. This experiment is representative of three independent experiments, each carried out on a different thymus.

Article Snippet: MAb against IL-1β (B-A15) was purchased from Diaclone Research (Besançon, France).

Techniques: Activity Assay, Isolation, Cell Culture, Incubation, Labeling, Derivative Assay, Binding Assay

Journal:

Article Title: High-Level Replication of Human Immunodeficiency Virus in Thymocytes Requires NF-?B Activation through Interaction with Thymic Epithelial Cells

doi:

Figure Lengend Snippet: TNF and, to a lesser extent, IL-1 induce NF-κB activity in thymocytes, and IL-7 is required for this effect. Whole-cell extracts were prepared from freshly isolated thymocytes (panel A, lane 1) or from thymocytes cultivated for 30, 60, or 45 h. (A) During the indicated culture times, thymocytes were either left untreated (lane 1, control for freshly isolated; lane 2, control 30 h; and lane 5, control 60 h) or were stimulated with either TNF (lanes 3 and 6), IL-1β (lanes 4 and 7), or TNF in the presence of TEC CM (TEC CM + TNF) (lane 9). TEC conditioned medium was also added alone (lane 8 [TEC CM]). The data shown here are representative of three independent experiments carried out on three thymuses. (B) Thymocytes were cultured for 45 h either untreated (lane 1, control 45 h), with TNF (lane 2), with TEC CM (lane 3), or with TNF in the presence of TEC CM (TEC CM + TNF) preincubated (lane 5) or not (lane 4) with antibody against IL-7. In lanes 6 to 8, the thymocytes were left untreated for 6 h (lane 6) or incubated with an antibody against IL-7Rα (lane 7) or with an IgG1 control serum (lane 8) before being cocultured with TEC for 45 h. The data shown here are representative of two independent experiments carried out on two thymuses. (C) Cells were left untreated for the 45 h of the culture (lane 1, control 45 h) or treated with IL-1β (lane 2), TNF (lane 3), IL-1β plus TNF plus IL-6 plus GM-CSF (lane 4), IL-7 (lane 5), IL-7 plus IL-1β (lane 6), or IL-7 plus TNF (lane 7). This experiment is representative of three independent experiments carried out on three thymuses. (D) Thymocytes were left untreated (lane 1, control) or stimulated with IL-7 (lane 2) or IL-7 in presence of anti-TNF and Il-1ra (lane 3). Whole-cell extracts were incubated with a 32P-labeled oligonucleotide representing κB motif. NF-κB activity is indicated. This experiment is representative of two independent experiments, each carried out on a different thymus.

Article Snippet: MAb against IL-1β (B-A15) was purchased from Diaclone Research (Besançon, France).

Techniques: Activity Assay, Isolation, Cell Culture, Incubation, Labeling

Journal: Pharmaceutics

Article Title: Elovanoids Counteract Inflammatory Signaling, Autophagy, Endoplasmic Reticulum Stress, and Senescence Gene Programming in Human Nasal Epithelial Cells Exposed to Allergens

doi: 10.3390/pharmaceutics14010113

Figure Lengend Snippet: ELVN-34 reduce the expression of pro-inflammatory cytokines, chemokines and cell adhesion molecule in HNEpC challenged with LPS or poly(I:C). HNEpC challenged with LPS or poly(I:C) (30 μg/mL) display enhanced production of pro-inflammatory cytokines and chemokines IL-6, IL-1β, IL-8/CXCL8, CCL2/MCP-1, CXCL1/KC/GRO, VEGF, and cell adhesion molecule ICAM1(CD54) compared to non-treated cells. This increase in the production of pro-inflammatory molecules is abrogated by the addition of ELVN-34 (500 nM) 30 min post-challenge. LPS and poly(I:C) induce a reduction of anti-inflammatory IL-10 expression in HNEpC that is restored to normal levels after treatment with ELVN-34 (500 nM). The results showed the averages of three independent experiments. (**** p < 0.0001, NS—not significant).

Article Snippet: In addition, the expression of the following cytokines was analyzed in the supernatant of HNEpC exposed to different allergic inductors: IL-6 (Catalog# 6802, Chondrex) range of detection: (9–600 pg/mL), IL-1β (Catalog# 6805, Chondrex) range of detection: (4–250 pg/mL) IL-8/CXCL8 (Quantikine ELISA Kit, Catalog# D8000C, R&D Systems) range of detection: (31–2000 pg/mL), vascular endothelial growth factor (VEGF) (Catalog# 6810, Chondrex) range of detection: (31–2000 pg/mL), ICAM1(CD54) (ELISA Kit, Catalog# ab100640, Abcam) range of detection: (16–1000 pg/mL), CXCL1/KC/GRO (Catalog# 6825, Chondrex) range of detection: CCL2/MCP-1 (Catalog# 6821, Chondrex) range of detection: (16–1000 pg/mL), and IL-10 (Catalog# 6806, Chondrex) range of detection: (8–500 pg/mL).

Techniques: Expressing

Journal: Pharmaceutics

Article Title: Elovanoids Counteract Inflammatory Signaling, Autophagy, Endoplasmic Reticulum Stress, and Senescence Gene Programming in Human Nasal Epithelial Cells Exposed to Allergens

doi: 10.3390/pharmaceutics14010113

Figure Lengend Snippet: HDM triggered multiple signaling in HNEpC. HNEpC challenged with HDM ( D. farinae + D. pteronyssinus ) (30 μg/mL) induces the expression of several genes related with autophagy (ATG3, ATG5, ATG7, BECLIN-1, and P62), unfolded protein response (UPR) (ATF6, CHOP, and IRE1), and matrix metalloproteinases (MMPs) (MMP8, MMP2, MMP9, MMP3, MMP12, TIMP1, and TIMP2). HDM stressors also induce the expression of senescence (P21, P16, P27, and P53) and inflammation genes (IL-1α, IL-6, and IL-1β) on HNEpC. The treatment with ELVN-34 Na (500 nM) reduces the expression of autophagy, UPR, MMP, senescence (except P53), and inflammation genes induced by HDM extracts. The results showed the averages of three independent experiments. (**** p < 0.0001, NS—not significant).

Article Snippet: In addition, the expression of the following cytokines was analyzed in the supernatant of HNEpC exposed to different allergic inductors: IL-6 (Catalog# 6802, Chondrex) range of detection: (9–600 pg/mL), IL-1β (Catalog# 6805, Chondrex) range of detection: (4–250 pg/mL) IL-8/CXCL8 (Quantikine ELISA Kit, Catalog# D8000C, R&D Systems) range of detection: (31–2000 pg/mL), vascular endothelial growth factor (VEGF) (Catalog# 6810, Chondrex) range of detection: (31–2000 pg/mL), ICAM1(CD54) (ELISA Kit, Catalog# ab100640, Abcam) range of detection: (16–1000 pg/mL), CXCL1/KC/GRO (Catalog# 6825, Chondrex) range of detection: CCL2/MCP-1 (Catalog# 6821, Chondrex) range of detection: (16–1000 pg/mL), and IL-10 (Catalog# 6806, Chondrex) range of detection: (8–500 pg/mL).

Techniques: Expressing