搏动性剪切应力导致人SH-SY5Y神经母细胞瘤细胞系中的DNA片段化。

Pulsatile shear stress leads to DNA fragmentation in human SH-SY5Y neuroblastoma cell line.

Triyoso D H, Good T A

Department of Chemical Engineering, Texas A&M University, College Station, TX 77843-3122, USA.

J Physiol. 1999 Mar 1;515 ( Pt 2)(Pt 2):355-65. doi: 10.1111/j.1469-7793.1999.355ac.x.

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.

使用剪切应力诱导细胞损伤的体外模型，我们证明对分化的人SH-SY5Y细胞施加剪切力会导致以DNA片段化为特征的细胞死亡。通过改良的锥板粘度计对细胞施加可控的剪切应力。2. 我们发现，通过对荧光素-12-dUTP缺口末端标记细胞进行流式细胞术测定，搏动性剪切应力导致45±4%的细胞出现DNA片段化。损伤后立即未观察到乳酸脱氢酶（LDH）释放；然而，损伤后24小时观察到显著的LDH释放。3. 经受搏动性剪切的细胞产生的一氧化氮比未剪切的对照细胞增加了两到三倍。4. 蛋白质合成、一氧化氮产生、Ca2+进入细胞以及百日咳毒素敏感的G蛋白激活的抑制作用减弱了剪切应力诱导的细胞损伤。5. 我们的结果首次表明，在体外对神经元样细胞施加搏动性剪切应力会导致一氧化氮依赖性细胞死亡。

Pulsatile shear stress leads to DNA fragmentation in human SH-SY5Y neuroblastoma cell line.