Cytosolic ionized calcium and bleb formation after acute cell injury of cultured rabbit renal tubule cells.
Cytosolic ionized calcium and bleb formation after acute cell injury of cultured rabbit renal tubule cells.
复制标题
DOI：
--
复制
发表时间：
1989-05
期刊：
影响因子：
--
通讯作者：
P. Phelps;M. Smith;B. Trump
中科院分区：
文献类型：
--
作者：
P. Phelps;M. Smith;B. Trump
本实验观察了体外培养的兔肾近曲小管细胞对多种损伤机制的反应，并观察了细胞内[Ca ~（2+）]i和细胞出泡的变化。[Ca2+]i在负载Fura 2的细胞中测量，并且通过相位显微镜观察起泡。损伤的严重程度通过电子显微镜进行评估，细胞杀伤通过台盼蓝染料摄取进行估计。损伤类型包括与巯基的相互作用（HgCl 2、N-乙基马来酰亚胺、对氯汞苯磺酸）、能量代谢的抑制（羰基氰间氯苯腙、KCN、KCN +碘乙酸）和离子失调（哇巴因、离子霉素、A23187）。细胞外钙离子（[Ca ~（2+）]_e）在损伤中的作用也进行了研究。HgCl 2、N-乙基马来酰亚胺和离子霉素+ [Ca 2 +]e引起[Ca 2 +]i的最高升高、最广泛的起泡和最快速的细胞死亡。对氯汞苯磺酸处理导致[Ca 2 +]i适度增加，以及不太广泛的起泡和较慢的细胞死亡。哇巴因和线粒体和细胞能量代谢的抑制剂仅引起[Ca 2 +]i增加2倍，一些水泡和延迟的细胞死亡。离子霉素- [Ca 2 +]e引起[Ca 2 +]i的短暂升高，最小的起泡和非常缓慢的细胞杀伤。[Ca 2 +]i的增加可能是由于细胞内储存的再分布（N-乙基马来酰亚胺、对氯汞苯磺酸、KCN、羰基氰间氯苯腙、离子霉素- [Ca 2 +]e）、细胞外[Ca 2 +]e的流入（离子霉素+ [Ca 2 +]e、哇巴因）或再分布和流入（HgCl 2）。因此，去除[Ca 2 +]e仅在某些类型的损伤中具有保护作用（HgCl 2，离子霉素）。在研究的所有类型的损伤中都观察到细胞质起泡，并且发生在细胞死亡之前。水泡形成迅速，扩大，有时脱离膜密封。我们的研究结果表明，细胞损伤，启动3倍或更大的持续升高[Ca 2 +]i，无论是由流入的[Ca 2 +]e或由Ca 2+释放从细胞内池，也与丰富的水泡形成和快速细胞死亡。
Changes in cytosolic calcium ([Ca2+]i) and cell blebbing of cultured rabbit kidney proximal tubule cells were studied in response to injury induced through a variety of mechanisms. [Ca2+]i was measured in Fura 2-loaded cells and blebbing was observed by phase microscopy. The severity of injury was evaluated by electron microscopy and cell killing was estimated by trypan blue dye uptake. The types of injury included interaction with sulfhydryl groups (HgCl2, N-ethylmaleimide, p-chloromercuribenzene sulfonic acid, inhibition of energy metabolism (carbonyl cyanide m-chlorophenylhydrazone, KCN, KCN + iodoacetate) and ion deregulation (ouabain, ionomycin, A23187). The role of extracellular calcium ([Ca2+]e) in injury was also studied. HgCl2, N-ethylmaleimide and ionomycin + [Ca2+]e caused the highest elevations of [Ca2+]i, the most extensive blebbing, and most rapid cell death. P-chloromercuribenzene sulfonic acid treatment resulted in a moderate increase in [Ca2+]i, as well as less extensive blebbing and slower cell death. Ouabain and inhibitors of mitochondrial and cellular energy metabolism caused only a 2-fold increase in [Ca2+]i, a few blebs and delayed cell death. Ionomycin - [Ca2+]e caused a transient elevation of [Ca2+]i, minimal blebbing and very slow cell killing. The increase in [Ca2+]i may result from redistribution of intracellular stores (N-ethylmaleimide, p-chloromercuribenzene sulfonic acid, KCN, carbonyl cyanide m-chlorophenylhydrazone, ionomycin - [Ca2+]e), from influx of extracellular [Ca2+]e (ionomycin + [Ca2+]e, ouabain), or from both redistribution and influx (HgCl2). Therefore, removing [Ca2+]e is protective only in certain types of injury, (HgCl2, ionomycin). Cytoplasmic blebbing was seen with all the types of injury studied and occurred before to cell death. Blebs formed rapidly, enlarged, and sometimes detached with membrane sealing. Our results indicate that cell injury which initiates a 3-fold or greater sustained elevation in [Ca2+]i, resulting from either an influx of [Ca2+]e or by Ca2+ release from intracellular pools, is also associated with abundant bleb formation and rapid cell death.