Watanabe S, Hirose M, Yasuda T, Miyazaki A, Sato N
Department of Gastroenterology, Juntendo University School of Medicine, Tokyo, Japan.
J Gastroenterol Hepatol. 1994 Jul-Aug;9(4):325-33. doi: 10.1111/j.1440-1746.1994.tb01250.x.
The aim of this study was to develop a new cellular restoration model using primary cultured rabbit gastric mucosal cells, and to evaluate the role of cytoskeleton and calmodulin in this process. Confluent monolayer mucosal cell sheets consisting mainly of mucous cells were wounded, and a rotating silicon tip was used to make cell-free areas of constant size. The process of restoration was monitored, and the cell-free area was measured and analysed quantitatively. Artificial wounds recovered in 36 h in controls; however, mucosal cell repair was inhibited by treatment with the actin inhibitor, cytochalasin B, and the calmodulin inhibitor, W-7. In the process of restoration, bromodeoxyuridine (BrdU)-positive cells appeared around the wound 24 h after injury and then disappeared after the complete repair. Because BrdU-positive cells were never detected within 24 h after wounding, initial recovery of the wound occurred only by cell migration. Both the migration and the proliferation stage are seen with this model. This culture model is suitable for the analysis of gastric mucosal restoration. Present data are consistent with the hypothesis that the integrity of the cytoskeletal system is important for restoration after damage of the gastric mucosa.
本研究的目的是利用原代培养的兔胃黏膜细胞建立一种新的细胞修复模型,并评估细胞骨架和钙调蛋白在此过程中的作用。主要由黏液细胞组成的汇合单层黏膜细胞片受到损伤,使用旋转硅尖制造大小恒定的无细胞区域。监测修复过程,并对无细胞区域进行定量测量和分析。对照组中人工伤口在36小时内愈合;然而,肌动蛋白抑制剂细胞松弛素B和钙调蛋白抑制剂W-7的处理抑制了黏膜细胞修复。在修复过程中,溴脱氧尿苷(BrdU)阳性细胞在损伤后24小时出现在伤口周围,然后在完全修复后消失。由于在受伤后24小时内从未检测到BrdU阳性细胞,伤口的初始恢复仅通过细胞迁移发生。该模型同时可见迁移和增殖阶段。这种培养模型适用于胃黏膜修复的分析。目前的数据与细胞骨架系统的完整性对胃黏膜损伤后修复很重要这一假设一致。