Abstract

The rapid healing of impaired intestinal surface plays a role in maintaining intestinal homeostasis. This study investigated the effect of calcium-sensing receptor (CaSR) on the migration and proliferation of intestinal porcine epithelial cells (IPEC-J2). Results showed that cell migration area and width were increased by R568 (CaSR activator) and decreased by NPS2143 (CaSR inhibitor). The protein level of GTP-rac1 and the phosphorylation of phospholipase C gamma 1 (PLCγ1) were increased by 2 µM R568. Furthermore, R568 + 120 µM NSC23766 (Rac1 inhibitor) and R568 + 1 µM U73122 (PLCγ1 inhibitor) decreased the protein level of GTP-rac1 and the phosphorylated PLCγ1, respectively, and both inhibited cell migration compared with R568. In addition, spermine increased the protein expression levels of CaSR and the levels of GTP-rac1 and the phosphorylated PLCγ1 and thereby promoted the migration of IPEC-J2 cells. Moreover, R568 improved the proliferation of the IPEC-J2 cells. Spermine increased cell proliferation, but NPS2143 incubated with spermine decreased cell proliferation compared with the spermine group. This study suggests that CaSR activation increased cell migration by activating Rac1 and PLCγ1 signaling and improved cell proliferation, and both effects were regulated by spermine by activating CaSR.

摘要

受损肠表面的快速愈合在维持肠稳态中发挥作用。本研究探讨了钙敏感受体 (CaSR) 对猪肠上皮细胞 (IPEC-J2) 迁移和增殖的影响。结果表明，R568（CaSR 激活剂）增加了细胞迁移面积和宽度，NPS2143（CaSR 抑制剂）减少了细胞迁移面积和宽度。GTP-rac1 的蛋白质水平和磷脂酶 C 伽马 1 (PLCγ1) 的磷酸化增加了 2 µM R568。此外，R568 + 120 µM NSC23766（Rac1 抑制剂）和 R568 + 1 µM U73122（PLCγ1 抑制剂）分别降低了 GTP-rac1 和磷酸化 PLCγ1 的蛋白质水平，并且与 R568 相比，两者都抑制了细胞迁移。此外，精胺增加CaSR的蛋白表达水平和GTP-rac1和磷酸化的PLCγ1的水平，从而促进IPEC-J2细胞的迁移。此外，R568 改善了 IPEC-J2 细胞的增殖。精胺增加细胞增殖，但与精胺组相比，与精胺一起孵育的 NPS2143 降低了细胞增殖。这项研究表明，CaSR 激活通过激活 Rac1 和 PLCγ1 信号传导增加细胞迁移并改善细胞增殖，并且这两种作用均受精胺通过激活 CaSR 调节。