PDE9 inhibition promotes proliferation of neural stem cells via cGMP-PKG pathway following oxygen-glucose deprivation/reoxygenation injury in vitro
PDE9 inhibition promotes proliferation of neural stem cells via cGMP-PKG pathway following oxygen-glucose deprivation/reoxygenation injury in vitro
复制标题
体外氧糖剥夺/复氧损伤后，PDE9 抑制可通过 cGMP-PKG 途径促进神经干细胞增殖。
DOI：
10.1016/j.neuint.2019.104630
复制
发表时间：
2020-02-01
影响因子：
4.2
通讯作者：
Jiang Qingsong
中科院分区：
文献类型：
--
作者：
Xiao Huan;Cheng Oumei;Jiang Qingsong
脑缺血是世界范围内导致死亡和长期残疾的主要原因之一。干细胞治疗有望为缺血性损伤后的结构和功能恢复提供一些有价值的策略。抑制磷酸二酯酶(PDEs)通过刺激神经干细胞(NSCs)的增殖而具有广谱的神经保护作用。然而，PDE9对脑缺血后NSCs增殖的潜在作用还没有得到很好的研究。本研究旨在评价PDE9抑制对神经干细胞增殖的影响，并探讨其抗脑缺血的作用机制。分别用CCK-8比色法和BrdU免疫荧光染色检测神经干细胞的存活和增殖情况。用酶联免疫试剂盒检测PDE9活性和cGMP水平。Western印迹法检测PKG和BDNF的蛋白表达。原代培养的海马神经干细胞暴露于缺氧-葡萄糖剥夺/复氧(OGD/R)可显著降低NSCs的存活率，但增加NSCs的增殖。同时，PDE9活性降低，cGMP水平升高，PKG和BDNF蛋白表达增加。与OGD/R模型组相比，PDE9抑制剂PF04447953提高了NSCs的存活率，PDE9活性进一步降低，NSCs增殖、cGMP水平、PKG和BDNF蛋白表达进一步增加。除PDE9活性和cGMP水平外，PF-04447953的上述作用可被PKG抑制剂KT5823逆转。综上所述，抑制PDE9可以促进OGD/R损伤后NSCs的增殖，这可能至少部分是通过cGMP-PKG途径实现的。
Cerebral ischemia is one of leading causes of death and long-term disability worldwide. Stem cell-based therapy is promising some valuable strategies for the structural and functional recovery after ischemic insult. The inhibition of phosphodiesterases (PDEs) has wide spectrum neuroprotective properties by stimulating proliferation of neural stem cells (NSCs). However, the potential role of PDE9 on NSCs proliferation after cerebral ischemia is not well investigated. The present study aimed to assess the contribution of PDE9 inhibition on the proliferation of NSCs and to determine the details of its underlying mechanisms against cerebral ischemia. The survival and proliferation of NSCs were assessed by CCK-8 assay and BrdU immunofluorescence staining, respectively. PDE9 activity and cGMP level were measured by ELISA kits. The protein expression of PKG and BDNF was detected by Western blot. Exposing NSCs of cultured primary hippocampus to oxygen-glucose deprivation/reoxygenation (OGD/R) significantly decreased the survival rate, but increased the proliferation of NSCs. Meanwhile, PDE9 activity was decreased, cGMP level was increased, PKG and BDNF protein expression was increased. PF04447953, a PDE9 inhibitor, increased the survival rate of NSCs, moreover, PDE9 activity reduced more, and NSCs proliferation, cGMP level, PKG and BDNF protein expression were increased further, compared with OGD/R model group. These effects of PF-04447953, except for PDE9 activity and cGMP level, were reversed by treatment with KT5823, a PKG inhibitor. Taken together, the inhibition of PDE9 can promote the proliferation of NSCs following OGD/R injury, which may be, at least partly, mediated by cGMP-PKG pathway.