机械应变对胃黏膜愈合过程中细胞迁移和增殖的影响：Rho 依赖性和 Rac 依赖性细胞骨架重组的作用
Effect of mechanical strain on gastric cellular migration and proliferation during mucosal healing: role of Rho dependent and Rac dependent cytoskeletal reorganisation.
作者信息
Osada T, Watanabe S, Tanaka H, Hirose M, Miyazaki A, Sato N
机构信息
Department of Gastroenterology, Juntendo University School of Medicine, Tokyo, Japan.
出版信息
Gut. 1999 Oct;45(4):508-15. doi: 10.1136/gut.45.4.508.
BACKGROUND
Various factors affect gastric wound healing. The influence of physical stimulation on gastric mucosal cells during the process of gastric wound healing is not completely understood.
AIMS
To assess the role of a physical stimulant, in this case mechanical strain, on gastric mucosal restoration.
METHODS
Mechanical strain was applied to adherent rat gastric mucosal cells (RGM1) cultured confluently on collagen type I coated silicone elastomer membrane in order to increase the dimension by an average of 5% and 10% at 5 cycles/minute for 72 hours after wounding. Repair of the wound was monitored every 12 hours for up to 72 hours using an inverted phase contrast microscope. Cell proliferation was detected using 5-bromodeoxyuridine staining. The cytoskeletal protein actin, a component of focal adhesion plaque protein, vinculin, and the small GTP-binding proteins RhoA and Rac1 were detected by immunohistochemistry in the cells located at the margin of and remote from the wound.
RESULTS
The cells located at the margin of the wound showed the greatest migration and proliferation and were found to express more rudimentary lamellipodia and filopodia in the absence of mechanical strain. Vinculin, RhoA, and Rac1 were also strongly expressed in the wound margin. Under conditions of mechanical strain, the speed of migration of cells slowed and fewer proliferating cells were detected around the wound in a strain strength dependent manner. Lamellipodial formation, vinculin, RhoA, and Rac1 were poorly expressed in the same area. However, in cells located more than 1 mm from the wound edge, cytoskeletal rearrangement, and the expression of vinculin, RhoA, and Rac1 were not influenced by mechanical strain.
CONCLUSIONS
Migration and proliferation of RGM1 cells in culture during wound healing were inhibited by mechanical strain, which caused dysfunction of the cytoskeletal and cell adhesion systems. It is suggested that physical stimulants such as mechanical strain may play a role in gastric wound healing in vivo by modifying cellular migration and proliferation.
背景
多种因素影响胃伤口愈合。在胃伤口愈合过程中，物理刺激对胃黏膜细胞的影响尚未完全明确。
目的
评估一种物理刺激物（在本研究中为机械牵张）对胃黏膜修复的作用。
方法
将机械牵张施加于在I型胶原包被的硅橡胶弹性体膜上汇合培养的贴壁大鼠胃黏膜细胞（RGM1），在损伤后以5次/分钟的频率使尺寸平均增加5%和10%，持续72小时。使用倒置相差显微镜每12小时监测一次伤口修复情况，直至72小时。采用5-溴脱氧尿苷染色检测细胞增殖。通过免疫组织化学检测位于伤口边缘和远离伤口处细胞中的细胞骨架蛋白肌动蛋白、粘着斑蛋白成分纽蛋白以及小GTP结合蛋白RhoA和Rac1。
结果
位于伤口边缘的细胞显示出最大的迁移和增殖，并且发现在无机械牵张的情况下表达更多原始的片状伪足和丝状伪足。纽蛋白、RhoA和Rac1在伤口边缘也强烈表达。在机械牵张条件下，细胞迁移速度减慢，并且在伤口周围检测到的增殖细胞数量减少，呈牵张强度依赖性。在同一区域，片状伪足形成、纽蛋白、RhoA和Rac1表达较差。然而，在距离伤口边缘超过1毫米的细胞中，细胞骨架重排以及纽蛋白、RhoA和Rac1的表达不受机械牵张影响。
结论
机械牵张抑制了培养的RGM1细胞在伤口愈合过程中的迁移和增殖，导致细胞骨架和细胞粘附系统功能障碍。提示机械牵张等物理刺激物可能通过改变细胞迁移和增殖在体内胃伤口愈合中发挥作用。
Zotero 插件