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Bioengineered Volume 12, 2021 - Issue 1
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Research Paper

The protective effects of naproxen against interleukin-1β (IL-1β)- induced damage in human umbilical vein endothelial cells (HUVECs)

Yuliang Wua Department of Cardiology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China
,
Ruina Haoa Department of Cardiology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China
,
Beidi Lanb Department of Structural Heart Disease, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China
,
Yiping Muc Department of Medical Information Management Office, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China
,
Fuping Dangd Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China
&
Ruitao Wangd Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, ChinaCorrespondence[email protected]
ORCID Iconhttps://orcid.org/0000-0002-1223-8732
ORCID Icon
Pages 5361-5372 | Received 31 Mar 2021, Accepted 10 Jul 2021, Published online: 24 Aug 2021

Figures & data

Figure 1. Cytotoxicity of naproxen in HUVECs. Cells were stimulated with naproxen at the concentrations of 1, 5, 10, 50, 100, and 500 μM for 24 h. (a). Molecular structure of naproxen; (b). Cell viability of HUVECs was measured using CCK-8 kit (#, ##, ###, P < 0.05, 0.01, 0.005 vs. vehicle group)

Figure 1. Cytotoxicity of naproxen in HUVECs. Cells were stimulated with naproxen at the concentrations of 1, 5, 10, 50, 100, and 500 μM for 24 h. (a). Molecular structure of naproxen; (b). Cell viability of HUVECs was measured using CCK-8 kit (#, ##, ###, P < 0.05, 0.01, 0.005 vs. vehicle group)

Figure 2. Naproxen prevents IL-1β-induced LDH release in HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). Morphology of HUVECs in different culture conditions; (b). LDH release was measured using a commercial kit (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 2. Naproxen prevents IL-1β-induced LDH release in HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). Morphology of HUVECs in different culture conditions; (b). LDH release was measured using a commercial kit (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 3. Naproxen suppresses IL-1β-induced expression of pro-inflammatory cytokines in HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). mRNA of IL-6 (1, 6.5, 4.6, 3.5); (b). mRNA of IL-12; (c). mRNA of TNF-α; (d). Secretions of IL-6; (e). Secretions of IL-12; (f). Secretions of TNF-α (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 3. Naproxen suppresses IL-1β-induced expression of pro-inflammatory cytokines in HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). mRNA of IL-6 (1, 6.5, 4.6, 3.5); (b). mRNA of IL-12; (c). mRNA of TNF-α; (d). Secretions of IL-6; (e). Secretions of IL-12; (f). Secretions of TNF-α (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 4. Naproxen inhibits IL-1β-induced expression of VEGF and tissue factor (TF) in HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). mRNA of VEGF; (b). mRNA of TF; (c). Production of VEGF; (d). Production of TF (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 4. Naproxen inhibits IL-1β-induced expression of VEGF and tissue factor (TF) in HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). mRNA of VEGF; (b). mRNA of TF; (c). Production of VEGF; (d). Production of TF (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 5. Naproxen reduces IL-1β-induced expression of ICAM-1 and E-selectin. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). mRNA of ICAM-1; (b). mRNA of E-selectin; (c). Production of ICAM-1; (d). Production of E-selectin (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 5. Naproxen reduces IL-1β-induced expression of ICAM-1 and E-selectin. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. (a). mRNA of ICAM-1; (b). mRNA of E-selectin; (c). Production of ICAM-1; (d). Production of E-selectin (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 6. Naproxen prevents IL-1β-induced attachment of THP-1 monocytes to HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. Attachment of THP-1 monocytes to HUVECs was measured using Calcein-AM staining (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 6. Naproxen prevents IL-1β-induced attachment of THP-1 monocytes to HUVECs. Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. Attachment of THP-1 monocytes to HUVECs was measured using Calcein-AM staining (###, P < 0.005 vs. vehicle group; *, **, P < 0.05, 0.01 vs. IL-1β group)

Figure 7. The protective effects of naproxen against IL-1β-induced attachment of THP-1 monocytes to HUVECs are dependent on KLF6. (a-b). Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. mRNA and protein levels of KLF6 were measured; (c-d). Cells were transfected with KLF6 siRNA, followed by stimulation with IL-1β (10 ng/mL) with or without naproxen (10 μM) for 24 h. Production of ICAM-1 and E-selectin was measured by ELISA. Attachment of THP-1 monocytes to HUVECs was measured using Calcein-AM staining (###, P < 0.005 vs. vehicle group; **, P < 0.01 vs. IL-1β group; $$, P < 0.01 vs. IL-1β+ IL-1β group)

Figure 7. The protective effects of naproxen against IL-1β-induced attachment of THP-1 monocytes to HUVECs are dependent on KLF6. (a-b). Cells were incubated with IL-1β (10 ng/mL) with or without naproxen (5, 10 μM) for 24 h. mRNA and protein levels of KLF6 were measured; (c-d). Cells were transfected with KLF6 siRNA, followed by stimulation with IL-1β (10 ng/mL) with or without naproxen (10 μM) for 24 h. Production of ICAM-1 and E-selectin was measured by ELISA. Attachment of THP-1 monocytes to HUVECs was measured using Calcein-AM staining (###, P < 0.005 vs. vehicle group; **, P < 0.01 vs. IL-1β group; $$, P < 0.01 vs. IL-1β+ IL-1β group)

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