OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms
Atherosclerosis is characterised by abnormal lipid and cell accumulation within arterial layers that leads to disturbed blood flow. Modified cholesterol forms such as oxidised low-density lipoprotein (oxLDL) enter cells altering their phenotype, triggering over-exuberant repair and arterial occlusio...
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| Format: | Article |
| Language: | English |
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Bioscientifica
2022-09-01
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| Series: | Vascular Biology |
| Subjects: | |
| Online Access: | https://vb.bioscientifica.com/view/journals/vb/4/1/VB-22-0009.xml |
| _version_ | 1798033044904345600 |
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| author | Majid Almansouri Pooja Patel Janet Chamberlain Sheila Francis |
| author_facet | Majid Almansouri Pooja Patel Janet Chamberlain Sheila Francis |
| author_sort | Majid Almansouri |
| collection | DOAJ |
| description | Atherosclerosis is characterised by abnormal lipid and cell accumulation within arterial layers that leads to disturbed blood flow. Modified cholesterol forms such as oxidised low-density lipoprotein (oxLDL) enter cells altering their phenotype, triggering over-exuberant repair and arterial occlusion, myocardial infarction or stroke. We hypothesised that oxLDL enters vascular wall cells and induces interleukin-1β (IL-1β) secretion, potentially via a caspase-1/NLRP3 mechanism. Human coronary artery endothelial cells (HCAEC) and smooth muscle cells (HCASMC), isolated from different donors, were cultured and stimulated (primed) with pro-inflammatory cytokines TNFα and IL-1α (10 ng/mL each, for 48 h), followed by incubation with human oxLDL (10–50 ug/mL) for up to 6 h. Inhibitors of caspase-1 (YVAD), NLRP3 (MCC950) and gasdermin D (disulfiram) were added 1 h before oxLDL. Cell lysates and culture supernatants were collected and analysed for IL-1β using ELISA. Microscopy imaging showed oxLDL entered stimulated cells and formed particles. OxLDL at 20 and 50 ug/mL induced the maximum release of IL-1β from stimulated HCASMCs and HCAECs, respectively, compared to control. Inhibition of either NLRP3, caspase-1 or gasdermin D significantly reduced the release of IL-1β (4-fold, P < 0.0001; 14-fold, P < 0.0001, 1.5-fold, P < 0.0003, respectively) in HCAEC. In contrast, in HCASMCs, only caspase-1 inhibition reduced the release of IL-1β (2.1-fold, P < 0.0001). HCAECs and HCASMCs elicited the release of IL-1β in response to the same stimulus via different mechanisms. In HCAECs, released IL-1β potentially exits via a GSDMD-induced membrane pore. These data suggest that caspase-1 or gasdermin D inhibition is likely to be effective vessel wall cell-specific strategies for the reduction of atherosclerosis. |
| first_indexed | 2024-04-11T20:23:58Z |
| format | Article |
| id | doaj.art-a213f6cd3c37453ca6b35addbd85a764 |
| institution | Directory Open Access Journal |
| issn | 2516-5658 |
| language | English |
| last_indexed | 2024-04-11T20:23:58Z |
| publishDate | 2022-09-01 |
| publisher | Bioscientifica |
| record_format | Article |
| series | Vascular Biology |
| spelling | doaj.art-a213f6cd3c37453ca6b35addbd85a7642022-12-22T04:04:43ZengBioscientificaVascular Biology2516-56582022-09-01411118https://doi.org/10.1530/VB-22-0009OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanismsMajid Almansouri0Pooja Patel1Janet Chamberlain2Sheila Francis3Department of Infection Immunity and Cardiovascular Disease, University of Sheffield, Medical School, Sheffield, UKDepartment of Infection Immunity and Cardiovascular Disease, University of Sheffield, Medical School, Sheffield, UKDepartment of Infection Immunity and Cardiovascular Disease, University of Sheffield, Medical School, Sheffield, UKDepartment of Infection Immunity and Cardiovascular Disease, University of Sheffield, Medical School, Sheffield, UK; Healthy Lifespan Institute HELSI, University of Sheffield, Medical School, Sheffield, UKAtherosclerosis is characterised by abnormal lipid and cell accumulation within arterial layers that leads to disturbed blood flow. Modified cholesterol forms such as oxidised low-density lipoprotein (oxLDL) enter cells altering their phenotype, triggering over-exuberant repair and arterial occlusion, myocardial infarction or stroke. We hypothesised that oxLDL enters vascular wall cells and induces interleukin-1β (IL-1β) secretion, potentially via a caspase-1/NLRP3 mechanism. Human coronary artery endothelial cells (HCAEC) and smooth muscle cells (HCASMC), isolated from different donors, were cultured and stimulated (primed) with pro-inflammatory cytokines TNFα and IL-1α (10 ng/mL each, for 48 h), followed by incubation with human oxLDL (10–50 ug/mL) for up to 6 h. Inhibitors of caspase-1 (YVAD), NLRP3 (MCC950) and gasdermin D (disulfiram) were added 1 h before oxLDL. Cell lysates and culture supernatants were collected and analysed for IL-1β using ELISA. Microscopy imaging showed oxLDL entered stimulated cells and formed particles. OxLDL at 20 and 50 ug/mL induced the maximum release of IL-1β from stimulated HCASMCs and HCAECs, respectively, compared to control. Inhibition of either NLRP3, caspase-1 or gasdermin D significantly reduced the release of IL-1β (4-fold, P < 0.0001; 14-fold, P < 0.0001, 1.5-fold, P < 0.0003, respectively) in HCAEC. In contrast, in HCASMCs, only caspase-1 inhibition reduced the release of IL-1β (2.1-fold, P < 0.0001). HCAECs and HCASMCs elicited the release of IL-1β in response to the same stimulus via different mechanisms. In HCAECs, released IL-1β potentially exits via a GSDMD-induced membrane pore. These data suggest that caspase-1 or gasdermin D inhibition is likely to be effective vessel wall cell-specific strategies for the reduction of atherosclerosis.https://vb.bioscientifica.com/view/journals/vb/4/1/VB-22-0009.xmlhcaecshcasmcsatherosclerosisoxldlcaspase-1gasdermin dil-1β |
| spellingShingle | Majid Almansouri Pooja Patel Janet Chamberlain Sheila Francis OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms Vascular Biology hcaecs hcasmcs atherosclerosis oxldl caspase-1 gasdermin d il-1β |
| title | OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms |
| title_full | OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms |
| title_fullStr | OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms |
| title_full_unstemmed | OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms |
| title_short | OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms |
| title_sort | oxldl induces the release of il 1β from primed human endothelial and smooth muscle cells via different caspase 1 dependent mechanisms |
| topic | hcaecs hcasmcs atherosclerosis oxldl caspase-1 gasdermin d il-1β |
| url | https://vb.bioscientifica.com/view/journals/vb/4/1/VB-22-0009.xml |
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