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Cadmium activates NLRP3 inflammasome-mediated pyroptosis in murine macrophage cell line RAW 264.7

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Caila Robinson

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Tampa

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Narasaiah Kolliputi

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Introduction: Cadmium (Cd) is an environmental toxin that affects the kidney, liver and lungs. Its effect on innate immunity is not well known.

Methods: RAW 264.7 macrophage cells were treated with different concentrations of CdCl₂ (5, 10, 15 μM) for 24h. Whole cell lysate was prepared, and western blot analysis was carried out for NLRP3, Caspase-1, and Gasdermin D. To detect pyroptosis, RAW cells were treated with 5, 10, 15 μM of CdCl₂, and Nigericin (positive control) followed by staining with FLICA reagent and Confocal microscopy.

Results: Western blot analysis revealed a higher expression of NLRP3, Caspase-1 and Gasdermin D in RAW 264.7 cells treated with CdClâ‚‚ relative to controls. Confocal imaging revealed an increased level of Caspase-1 activity in CdClâ‚‚ treated cells relative to controls.

Conclusion: The data shows that Cadmium induces NLRP3 inflammasome-mediated Caspase-1 activity resulting in pyroptosis induced cell death in RAW 264.7 macrophage cells.

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Cadmium activates NLRP3 inflammasome-mediated pyroptosis in murine macrophage cell line RAW 264.7

Introduction: Cadmium (Cd) is an environmental toxin that affects the kidney, liver and lungs. Its effect on innate immunity is not well known.

Methods: RAW 264.7 macrophage cells were treated with different concentrations of CdCl₂ (5, 10, 15 μM) for 24h. Whole cell lysate was prepared, and western blot analysis was carried out for NLRP3, Caspase-1, and Gasdermin D. To detect pyroptosis, RAW cells were treated with 5, 10, 15 μM of CdCl₂, and Nigericin (positive control) followed by staining with FLICA reagent and Confocal microscopy.

Results: Western blot analysis revealed a higher expression of NLRP3, Caspase-1 and Gasdermin D in RAW 264.7 cells treated with CdClâ‚‚ relative to controls. Confocal imaging revealed an increased level of Caspase-1 activity in CdClâ‚‚ treated cells relative to controls.

Conclusion: The data shows that Cadmium induces NLRP3 inflammasome-mediated Caspase-1 activity resulting in pyroptosis induced cell death in RAW 264.7 macrophage cells.