Qi Da-Shi, Tao Jin-Hao, Zhang Lian-Qin, Li Man, Wang Mei, Qu Rui, Zhang Shi-Chun, Liu Pei, Liu Fuming, Miu Jian-Cheng, Ma Jing-Yi, Mei Xin-Yu, Zhang Fayong
Jiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application Technology, Xuzhou Medical University, People's Republic of China; Department of Genetics, Xuzhou Medical University, Xuzhou, Jiangsu 221004, People's Republic of China.
Pediatric Emergency and Critical Care Center, Children' Hospital of Fudan University, Shanghai, People's Republic of China.
Brain Res. 2016 Dec 15;1653:67-74. doi: 10.1016/j.brainres.2016.10.017. Epub 2016 Oct 18.
Cilostazol(CTL) is a phosphodiesterase inhibitor, which has been widely used as anti-platelet agent. It also has preventive effects on various central nervous system (CNS) diseases, including ischemic stroke, Parkinson's disease and Alzheimer disease. However, the molecular mechanism underlying the protective effects of CTL is still unclear, and whether CTL can prevent I/R induced cognitive deficit has not been reported. Transient global brain ischemia was induced by 4-vessel occlusion in adult male Sprague-Dawley rats. The open field tasks and Morris water maze were used to assess the effect of CTL on anxiety-like behavioral and cognitive impairment after I/R. Western blotting were performed to examine the expression of related proteins, and HE-staining was used to detect the percentage of neuronal death in the hippocampal CA1 region. Here we found that CTL significantly improved cognitive deficits and the behavior of rats in Morris water maze and open field tasks (P<0.05). HE staining results showed that CTL could significantly protect CA1 neurons against cerebral I/R (P<0.05). Additionally, Akt1 phosphorylation levels were evidently up-regulated (P<0.05), while the activation of JNK3, which is an important contributor to I/R-induced neuron apoptosis, was reduced by CTL after I/R (P<0.05), and caspase-3 levels were also decreased by CTL treatment. Furthermore, all of CTL's protective effects were reversed by LY294002, which is a PI3K/Akt1 inhibitor. Taken together, our results suggest that CTL could protect hippocampal neurons and ameliorate the impairment of learning/memory abilities and locomotor/ exploratory activities in ischemic stroke via a PI3K-Akt1/JNK3/caspase-3 dependent mechanism.
西洛他唑（CTL）是一种磷酸二酯酶抑制剂，已被广泛用作抗血小板药物。它对包括缺血性中风、帕金森病和阿尔茨海默病在内的各种中枢神经系统（CNS）疾病也有预防作用。然而，CTL保护作用的分子机制仍不清楚，且CTL是否能预防缺血/再灌注（I/R）诱导的认知缺陷尚未见报道。采用四血管闭塞法诱导成年雄性Sprague-Dawley大鼠短暂性全脑缺血。利用旷场试验和莫里斯水迷宫来评估CTL对I/R后焦虑样行为和认知障碍的影响。进行蛋白质免疫印迹法检测相关蛋白的表达，并用苏木精-伊红（HE）染色检测海马CA1区神经元死亡的百分比。在此我们发现，CTL显著改善了大鼠在莫里斯水迷宫和旷场试验中的认知缺陷及行为表现（P<0.05）。HE染色结果显示，CTL能显著保护CA1神经元免受脑I/R损伤（P<0.05）。此外，Akt1磷酸化水平明显上调（P<0.05），而I/R后CTL可降低对I/R诱导的神经元凋亡起重要作用的JNK3的激活（P<0.05），CTL处理也降低了半胱天冬酶-3水平。此外，LY294002（一种PI3K/Akt1抑制剂）可逆转CTL的所有保护作用。综上所述，我们的结果表明，CTL可通过PI3K-Akt1/JNK3/半胱天冬酶-3依赖性机制保护海马神经元，并改善缺血性中风中学习/记忆能力及运动/探索活动的损伤。
