Mo Weiwei, Liu Guangmang, Wu Caimei, Jia Gang, Zhao Hua, Chen Xiaoling, Wang Jing
Institute of Animal Nutrition, Sichuan Agricultural University, Chengdu, China.
Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Chengdu, China.
Anim Biotechnol. 2022 Dec;33(7):1492-1503. doi: 10.1080/10495398.2021.1910044. Epub 2021 Apr 18.
Intestinal epithelial restitution is partly dependent on cell migration, which reseals superficial wounding after injury. Here, we tested the hypothesis that stromal interaction molecule 1(STIM1) regulates porcine intestinal epithelial cell migration by activating transient receptor potential canonical 1 (TRPC1) signaling. Results showed that the knockdown of STIM1 repressed cell migration after wounding, reduced the protein concentration of STIM1 and TRPC1, and decreased the inositol trisphosphate (IP3) content in IPEC-J2 cells ( < 0.05). However, overexpression of STIM1 obtained opposite results ( < 0.05). The inhibition of TRPC1 activity by treatment with SKF96365 in cells overexpressing wild-type and mutant STIM1 attenuated the STIM1 overexpression-induced increase of cell migration, STIM1, TRPC1 and IP3 ( < 0.05). In addition, polyamine depletion caused by α-difluoromethylornithine (DFMO) resulted in the decrease of above-mentioned parameters, and exogenous polyamine could attenuate the negative effects of DFMO on IPEC-J2 cells ( < 0.05). Moreover, the overexpression of STIM1 could rescue cell migration, the protein level of STIM1 and TRPC1, and IP3 content in polyamine-deficient IPEC-J2 cells ( < 0.05). These results indicated that STIM1 could enhance porcine intestinal epithelial cell migration via the TRPC1 signaling pathway. Inhibition of cell migration by polyamine depletion resulted from the reduction of STIM1 activity.
肠上皮修复部分依赖于细胞迁移，细胞迁移可在损伤后重新封闭表面伤口。在此，我们验证了基质相互作用分子1（STIM1）通过激活瞬时受体电位香草酸亚型1（TRPC1）信号通路来调节猪肠上皮细胞迁移的假说。结果显示，敲低STIM1可抑制伤口愈合后的细胞迁移，降低IPEC-J2细胞中STIM1和TRPC1的蛋白浓度，并降低肌醇三磷酸（IP3）含量（<0.05）。然而，过表达STIM1则得到相反的结果（<0.05）。用SKF96365处理过表达野生型和突变型STIM1的细胞，抑制TRPC1活性，可减弱STIM1过表达诱导的细胞迁移、STIM1、TRPC1和IP3的增加（<0.05）。此外，α-二氟甲基鸟氨酸（DFMO）导致的多胺耗竭使上述参数降低，而外源性多胺可减弱DFMO对IPEC-J2细胞的负面影响（<0.05）。此外，过表达STIM1可挽救多胺缺乏的IPEC-J2细胞的细胞迁移、STIM1和TRPC1的蛋白水平以及IP3含量（<0.05）。这些结果表明，STIM1可通过TRPC1信号通路增强猪肠上皮细胞迁移。多胺耗竭导致的细胞迁移抑制是由于STIM1活性降低所致。
