Journal: Frontiers in Pharmacology

Article Title: Dauricine Attenuates Vascular Endothelial Inflammation Through Inhibiting NF-κB Pathway

doi: 10.3389/fphar.2021.758962

Figure Lengend Snippet: Chemical structure and cytotoxicity induced by dauricine. (A) Chemical structure of dauricine. (B) Dauricine was safe to use for HUVECs at concentrations of 0–40 μM, as determined by MTT assay (n = 5). HUVECs, human umbilical vein endothelial cells.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were cultured in specific endothelial medium (ScienCell, Carlsbad, CA, United States) containing 1% endothelial cell growth supplement, 5% fetal bovine serum, and 1% penicillin-streptomycin solution in a cell incubator (37°C with 5% CO 2 ).

Techniques: MTT Assay

Journal: Frontiers in Pharmacology

Article Title: Dauricine Attenuates Vascular Endothelial Inflammation Through Inhibiting NF-κB Pathway

doi: 10.3389/fphar.2021.758962

Figure Lengend Snippet: Dauricine inhibited monocyte–endothelial cell interaction. (A) Representative images of THP-1 cells (green) that adhered to HUVECs (scale bar: 100 μm). (B) Quantitative analysis of THP-1 cells that adhered to HUVECs. Data are expressed as mean ± SD. ### p < 0.001 versus the vehicle group; *** p < 0.001 versus the vehicle + IL-1β group. DMSO, dimethyl sulfoxide; IL-1β, interleukin-1β; HUVECs, human umbilical vein endothelial cells.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were cultured in specific endothelial medium (ScienCell, Carlsbad, CA, United States) containing 1% endothelial cell growth supplement, 5% fetal bovine serum, and 1% penicillin-streptomycin solution in a cell incubator (37°C with 5% CO 2 ).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Dauricine Attenuates Vascular Endothelial Inflammation Through Inhibiting NF-κB Pathway

doi: 10.3389/fphar.2021.758962

Figure Lengend Snippet: Dauricine decreased the expression of ICAM-1, VCAM-1, and E-selectin in HUVECs. (A) Relative mRNA expression levels of ICAM-1, VCAM-1, and E-selectin in HUVECs treated with IL-1β and dauricine by quantitative PCR (qPCR). (B) The protein levels of ICAM-1, VCAM-1, and E-selectin in HUVECs by Western blotting. (C) Quantification of the relative expression level of proteins in panel B. Data are expressed as mean ± SD. ### p < 0.001 versus the vehicle group; *** p < 0.001, ** p < 0.01, * p < 0.05 versus the vehicle + IL-1β group. HUVECs, human umbilical vein endothelial cells; IL-1β, interleukin-1β.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were cultured in specific endothelial medium (ScienCell, Carlsbad, CA, United States) containing 1% endothelial cell growth supplement, 5% fetal bovine serum, and 1% penicillin-streptomycin solution in a cell incubator (37°C with 5% CO 2 ).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Dauricine Attenuates Vascular Endothelial Inflammation Through Inhibiting NF-κB Pathway

doi: 10.3389/fphar.2021.758962

Figure Lengend Snippet: Dauricine inhibited IL-1β-induced NF-κB activation. (A) Phosphorylation of p65 and IκBα in HUVECs treated with IL-1β and dauricine at different concentrations. (B) Quantification of the phosphorylation of p65 and IκBα displayed in panel A. ## p < 0.01, # p < 0.05 versus the vehicle group; * p < 0.05 versus the vehicle + IL-1β group. (C) Nuclear translocation of p65 as indicated by the immunofluorescence in HUVECs treated with IL-1β and dauricine (scale bar: 200 μm). DAPI was used for nucleus staining (blue). P65 (green) was stained with a secondary antibody of Alexa Fluor 488. (D) Cytoplasmic and nuclear distribution of p65 in HUVECs as detected by Western blotting analysis. IL-1β, interleukin-1β; HUVECs, human umbilical vein endothelial cells.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were cultured in specific endothelial medium (ScienCell, Carlsbad, CA, United States) containing 1% endothelial cell growth supplement, 5% fetal bovine serum, and 1% penicillin-streptomycin solution in a cell incubator (37°C with 5% CO 2 ).

Techniques: Activation Assay, Phospho-proteomics, Translocation Assay, Immunofluorescence, Staining, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Dauricine Attenuates Vascular Endothelial Inflammation Through Inhibiting NF-κB Pathway

doi: 10.3389/fphar.2021.758962

Figure Lengend Snippet: Dauricine attenuated LPS-induced acute lung injury in vivo . (A) Survival curves for mice treated with LPS and dauricine (20 mg/kg), n = 10/group. (B) Protein levels of ICAM-1, VCAM-1, and E-selectin and the phosphorylation of IκBα and p65 in lung tissues. (C) Quantification analysis of the protein levels in panel B. ### p < 0.001, ## p < 0.01, # p < 0.05 versus the vehicle group; ** p < 0.01, * p < 0.05 versus the vehicle + LPS group. (D) Level of endothelial cell-expressed ICAM-1 in lung tissues, as detected by immunofluorescence assay (scale bar: 20 μm). DAPI was used for nucleus staining (blue), ICAM-1 was stained red, and CD31 was stained green. (E) Nuclear translocation of p65 in endothelial cells as detected by immunofluorescence assay in lung tissues (scale bar: 10 μm). DAPI was used for nucleus staining (blue), p65 was stained red, and CD31 was stained green. (F) Quantification analysis of ICAM-1 in panel D. ## p < 0.01 versus the vehicle group; * p < 0.05 versus the vehicle + LPS group. (G) Quantification analysis of nucleic p65 in panel E. ### p < 0.001 versus the vehicle group; * p < 0.05 versus the vehicle + LPS group. LPS, lipopolysaccharide.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were cultured in specific endothelial medium (ScienCell, Carlsbad, CA, United States) containing 1% endothelial cell growth supplement, 5% fetal bovine serum, and 1% penicillin-streptomycin solution in a cell incubator (37°C with 5% CO 2 ).

Techniques: In Vivo, Phospho-proteomics, Immunofluorescence, Staining, Translocation Assay

Journal: Frontiers in Pharmacology

Article Title: Dauricine Attenuates Vascular Endothelial Inflammation Through Inhibiting NF-κB Pathway

doi: 10.3389/fphar.2021.758962

Figure Lengend Snippet: Schematic illustration of the anti-inflammatory effect of dauricine. Proinflammatory stimulus such as LPS and IL-1β increased the phosphorylation and nuclear translocation of the NF-κB p65 subunit, which resulted in the upregulation of adhesion molecules and many other inflammatory cytokines that could aggravate vascular inflammation. Dauricine showed an inhibition of NF-kB pathway and downregulated the expression of its downstream genes to attenuate endothelial inflammation. IL-1β, interleukin-1β; LPS, lipopolysaccharide.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were cultured in specific endothelial medium (ScienCell, Carlsbad, CA, United States) containing 1% endothelial cell growth supplement, 5% fetal bovine serum, and 1% penicillin-streptomycin solution in a cell incubator (37°C with 5% CO 2 ).

Techniques: Phospho-proteomics, Translocation Assay, Inhibition, Expressing