Pulsatile shear stress leads to DNA fragmentation in human SH-SY5Y neuroblastoma cell line.
1. Using an in vitro model of shear stress-induced cell injury we demonstrate that application of shear to differentiated human SH-SY5Y cells leads to cell death characterized by DNA fragmentation. Controlled shear stress was applied to cells via a modified cone and plate viscometer. 2. We show that pulsatile shear stress leads to DNA fragmentation, as determined via flow cytometry of fluorescein-12-dUTP nick-end labelled cells, in 45 +/- 4 % of cells. No lactate dehydrogenase (LDH) release was observed immediately after injury; however, 24 h after injury significant LDH release was observed. 3. Nitric oxide production by cells subjected to pulsatile shear increased two- to threefold over that in unsheared control cells. 4. Inhibition of protein synthesis, nitric oxide production, Ca2+ entry into cells, and pertussis toxin-sensitive G protein activation attenuated the shear stress-induced cell injury. 5. Our results show for the first time that application of pulsatile shear stress to a neuron-like cell in vitro leads to nitric oxide-dependent cell death.
Department of Chemical Engineering, Texas A&M University, College Station, TX 77843-3122, USA.
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Triyoso
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D002118:Calcium / Q000378:metabolism; D053938:DNA Fragmentation / Q000502:physiology*; D005434:Flow Cytometry; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000502:physiology; D006801:Humans; D020287:In Situ Nick-End Labeling; D009474:Neurons / Q000502:physiology*; D009569:Nitric Oxide / Q000096:biosynthesis; D037342:Pertussis Toxin; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011673:Pulsatile Flow; D013314:Stress, Mechanical; D014407:Tumor Cells, Cultured / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D011500:Protein Synthesis Inhibitors; D010566:Virulence Factors, Bordetella; D009569:Nitric Oxide; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D002118:Calcium