University Grenoble-Alpes, CEDEX 9, 38043 Grenoble, France.
Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Laboratory of Biology & Biotechnology for Health, Interdisciplinary Research Institute of Grenoble, 38000 Grenoble, France.
Int J Mol Sci. 2023 Oct 21;24(20):15428. doi: 10.3390/ijms242015428.
Prokineticins are a family of small proteins with diverse roles in various tissues, including the brain. However, their specific effects on different cerebral cell types and blood-brain barrier (BBB) function remain unclear. The aim of this study was to investigate the effects of PROK1 and PROK2 on murine cerebral cell lines, bEnd.3, C8.D30, and N2a, corresponding to microvascular endothelial cells, astrocytes and neurons, respectively, and on an established BBB co-culture model. Western blot analysis showed that prokineticin receptors (PROKR1 and PROKR2) were differentially expressed in the considered cell lines. The effect of PROK1 and PROK2 on cell proliferation and migration were assessed using time-lapse microscopy. PROK1 decreased neural cells' proliferation, while it had no effect on the proliferation of endothelial cells and astrocytes. In contrast, PROK2 reduced the proliferation of all cell lines tested. Both PROK1 and PROK2 increased the migration of all cell lines. Blocking PROKRs with the PROKR1 antagonist (PC7) and the PROKR2 antagonist (PKR-A) inhibited astrocyte PROK2-mediated migration. Using the insert co-culture model of BBB, we demonstrated that PROKs increased BBB permeability, which could be prevented by PROKRs' antagonists.
趋化因子是一类具有多种功能的小蛋白家族，在包括大脑在内的各种组织中都有表达。然而，它们对不同脑内细胞类型和血脑屏障（BBB）功能的具体影响尚不清楚。本研究旨在探讨 PROK1 和 PROK2 对分别代表微血管内皮细胞、星形胶质细胞和神经元的小鼠脑细胞系 bEnd.3、C8.D30 和 N2a 以及已建立的 BBB 共培养模型的影响。Western blot 分析显示，趋化因子受体（PROKR1 和 PROKR2）在考虑的细胞系中差异表达。使用延时显微镜评估 PROK1 和 PROK2 对细胞增殖和迁移的影响。PROK1 降低了神经细胞的增殖，而对内皮细胞和星形胶质细胞的增殖没有影响。相比之下，PROK2 降低了所有测试细胞系的增殖。PROK1 和 PROK2 均增加了所有细胞系的迁移。用 PROKR1 拮抗剂（PC7）和 PROKR2 拮抗剂（PKR-A）阻断 PROKRs 可抑制星形胶质细胞 PROK2 介导的迁移。通过 BBB 的插入共培养模型，我们证明 PROKs 增加了 BBB 的通透性，而 PROKRs 的拮抗剂可以预防这种通透性增加。
