GABA 受体拮抗剂促进急性脑缺血后小鼠海马神经发生和认知功能恢复。

GABA receptor antagonist promotes hippocampal neurogenesis and facilitates cognitive function recovery following acute cerebral ischemia in mice.

机构信息

Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.

Laboratory Research Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.

出版信息

Stem Cell Res Ther. 2021 Jan 7;12(1):22. doi: 10.1186/s13287-020-02059-x.

PURPOSE AND BACKGROUND

Previous studies have suggested that promoting endogenous neurogenesis has great significance for the recovery of cognitive dysfunction caused by cerebral ischemia (CI). Pharmacological inhibition of GABA receptor can enhance neurogenesis in adult healthy and depressed mice. In the study, we intended to investigate the effects of GABA receptor antagonists on cognitive function and hippocampal neurogenesis in mice following CI.

METHODS

Adult mice were subjected to bilateral common carotid artery occlusion (BCCAO) for 20 min to induce CI and treated with CGP52432 (antagonist of GABA receptor, CGP, 10 mg/kg intraperitoneal injection) starting 24 h after CI. The Morris water maze test was performed to test spatial learning and memory at day 28. Immunofluorescence was applied to detect neurogenesis in the DG region at day 14 and 28. In in vitro experiments, cell proliferation was detected by CCK8 and immunofluorescence, and the expression of cAMP/CREB signaling pathway-related proteins was detected by ELISA assay and Western blot.

RESULTS

CGP significantly improved spatial learning and memory disorders caused by CI, and it enhanced the proliferation of neural stem cells (NSCs), the number of immature neurons, and the differentiation from newborn cells to neurons. In vitro experiments further confirmed that CGP dose-dependently enhanced the cell viability of NSCs, and immunofluorescence staining showed that CGP promoted the proliferation of NSCs. In addition, treatment with CGP increased the expression of cAMP, PKA, and pCREB in cultured NSCs.

CONCLUSION

Inhibition of GABA receptor can effectively promote hippocampal neurogenesis and improve spatial learning and memory in adult mice following CI.

目的和背景

先前的研究表明，促进内源性神经发生对于脑缺血（CI）引起的认知功能障碍的恢复具有重要意义。GABA 受体的药理学抑制可以增强成年健康和抑郁小鼠的神经发生。在这项研究中，我们旨在研究 GABA 受体拮抗剂对 CI 后小鼠认知功能和海马神经发生的影响。

方法

成年小鼠接受双侧颈总动脉闭塞（BCCAO）20 分钟以诱导 CI，并在 CI 后 24 小时开始用 CGP52432（GABA 受体拮抗剂，CGP，腹腔注射 10mg/kg）进行治疗。在第 28 天进行 Morris 水迷宫测试以测试空间学习和记忆。免疫荧光法用于检测第 14 天和第 28 天 DG 区的神经发生。在体外实验中，通过 CCK8 和免疫荧光法检测细胞增殖，通过 ELISA 测定和 Western blot 检测 cAMP/CREB 信号通路相关蛋白的表达。

结果

CGP 显著改善了 CI 引起的空间学习和记忆障碍，增强了神经干细胞（NSCs）的增殖、未成熟神经元的数量以及从新生细胞向神经元的分化。体外实验进一步证实，CGP 剂量依赖性地增强了 NSCs 的细胞活力，免疫荧光染色显示 CGP 促进了 NSCs 的增殖。此外，CGP 处理增加了培养的 NSCs 中 cAMP、PKA 和 pCREB 的表达。