Crystal-cell interaction and apoptosis in oxalate-associated injury of renal epithelial cells.
Crystal-cell interaction and apoptosis in oxalate-associated injury of renal epithelial cells.
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发表时间：
1999-11
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通讯作者：
Saeed R. Khan;K. Byer;Sivagnanan Thamilselvan;R. Hackett;Wayne T. McCormack;Neal A. Benson;Karen L. Vaughn;Gregory W. Erdos
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作者：
Saeed R. Khan;K. Byer;Sivagnanan Thamilselvan;R. Hackett;Wayne T. McCormack;Neal A. Benson;Karen L. Vaughn;Gregory W. Erdos
两种肾上皮细胞系LLC-PK1和Madin-Darby犬肾(MDCK)在单层培养中，暴露于草酸(Ox)和/或草酸钙(CaOx)晶体中，以观察细胞对这些挑战的反应。此外，将LLC-PK1细胞暴露于高浓度的Ox不同时间段，以探讨细胞凋亡在Ox相关细胞损伤中的作用。这两种类型的细胞在暴露于Ox时都显示出损坏的迹象。然而，LLC-PK1细胞比MDCK细胞更敏感。单层细胞乳酸脱氢酶释放量显著增加，台盼蓝拒染率明显降低。最明显的是细胞从底物上脱落。细胞暴露在CaOx晶体中，导致它们附着在细胞表面，随后内化。用流式细胞术定量细胞凋亡量，用透射电子显微镜观察细胞形态，用凝胶电泳法检测DNA梯状条带，观察到明显的细胞凋亡改变，包括核染色质凝集、边集、DNA断裂、质膜磷脂酰丝氨酸从细胞内向细胞表面迁移等。然而，这些细胞也出现了一些坏死性改变，如质膜完整性丧失和乳酸脱氢酶释放，表明细胞的凋亡过程被中断。
Two renal epithelial cell lines, LLC-PK1 and Madin-Darby canine kidney (MDCK), were grown in monolayers and exposed to oxalate (Ox) and/or calcium oxalate (CaOx) crystals to investigate cellular responses to these challenges. In addition, LLC-PK1 cells were exposed to high concentrations of Ox for various time periods to investigate the role of apoptosis in Ox-associated cell injury. Both cell types showed signs of damage when exposed to Ox. However, LLC-PK1 cells appeared more sensitive than MDCK cells. There was a significant increase in release of lactate dehydrogenase into the medium and decrease in trypan blue exclusion by cells in the monolayer. Most noticeable was the detachment of cells from the substrate. Exposure of cells to CaOx crystals resulted in their attachment to cell surfaces followed by internalization. Using flow cytometry for quantification of apoptotic cells, transmission electron microscopy for morphology, and electrophoresis for DNA laddering detection, we observed significant apoptotic changes including condensation and margination of nuclear chromatin, DNA fragmentation, and migration of phosphatidylserine of the plasma membrane from inside to the cell surface. However, these cells also showed some necrotic changes such as loss of plasma membrane integrity and release of lactate dehydrogenase, indicating that the apoptotic process was interrupted.