Abstract：
Postoperative cognitive dysfunction (POCD) is characterized by impaired learning and memory. 6 h duration isoflurane anesthesia is an important factor to induce POCD, and the dysfunction of ryanodine receptor (RyR) in the hippocampus may be involved in this process. We investigated the expression of RyR3 in the hippocampus of mice after 6-h duration isoflurane anesthesia, as well as the improvement of RyR receptor agonist caffeine on POCD mice, while attempting to identify the underlying molecular mechanism.
We constructed a POCD model using 8-week-old male C57BL/6J mice that were exposed to 6-h duration isoflurane. Prior to the three-day cognitive behavioral experiment, RyR agonist caffeine were injected. Fear conditioning and location memory tests were used in behavioral studies. We also exposed the mouse neuroblastoma cell line Neuro-2a (N2A) to 6-h duration isoflurane exposure to simulate the conditions of in vivo cognitive dysfunction. We administered ryanodine receptor agonist (caffeine) and inhibitor (ryanodine) to N2a cells. Following that, we performed a series of bioinformatics analysis to discover proteins that are involved in the development of cognitive dysfunction. Rt-PCR and Western blot were used to assess mRNA level and protein expression.
6-h duration isoflurane anesthesia induced cognitive dysfunction and increased RyR3 mRNA levels in hippocampus. The mRNA levels of RyR3 in cultured N2a cells after anesthesia were comparable to those in vivo, and the RyR agonist caffeine corrected the expression of some cognitive-related phenotypic proteins that were disturbed after anesthesia. Intraperitoneal injection of RyR agonist caffeine can improve cognitive function after isoflurane anesthesia in mice, and bioinformatics analyses suggest that CaMKⅣ may be involved in the molecular mechanism.
Ryanodine receptor agonist caffeine may improve cognitive dysfunction in mice after isoflurane anesthesia.
We constructed a POCD model using 8-week-old male C57BL/6J mice that were exposed to 6-h duration isoflurane. Prior to the three-day cognitive behavioral experiment, RyR agonist caffeine were injected. Fear conditioning and location memory tests were used in behavioral studies. We also exposed the mouse neuroblastoma cell line Neuro-2a (N2A) to 6-h duration isoflurane exposure to simulate the conditions of in vivo cognitive dysfunction. We administered ryanodine receptor agonist (caffeine) and inhibitor (ryanodine) to N2a cells. Following that, we performed a series of bioinformatics analysis to discover proteins that are involved in the development of cognitive dysfunction. Rt-PCR and Western blot were used to assess mRNA level and protein expression.
6-h duration isoflurane anesthesia induced cognitive dysfunction and increased RyR3 mRNA levels in hippocampus. The mRNA levels of RyR3 in cultured N2a cells after anesthesia were comparable to those in vivo, and the RyR agonist caffeine corrected the expression of some cognitive-related phenotypic proteins that were disturbed after anesthesia. Intraperitoneal injection of RyR agonist caffeine can improve cognitive function after isoflurane anesthesia in mice, and bioinformatics analyses suggest that CaMKⅣ may be involved in the molecular mechanism.
Ryanodine receptor agonist caffeine may improve cognitive dysfunction in mice after isoflurane anesthesia.
摘要：
背景：术后认知功能障碍（POCD）的特征是学习和记忆受损。持续6h的异氟烷麻醉是诱发POCD的重要因素,海马ryanodine受体（RyR）的功能障碍可能参与了这一过程。我们研究了RyR3在6小时持续异氟醚麻醉后小鼠海马中的表达，以及RyR受体激动剂咖啡因对POCD小鼠的改善，同时试图确定潜在的分子机制。材料：我们使用暴露于6小时持续异氟烷的8周龄雄性C57BL/6小鼠构建POCD模型。在为期三天的认知行为实验之前，注射RyR激动剂咖啡因。行为研究中使用了恐惧条件和位置记忆测试。我们还将小鼠神经母细胞瘤细胞系Neuro-2a（N2A）暴露于持续6小时的异氟烷暴露，以模拟体内认知功能障碍的条件。我们向N2a细胞施用ryanodine受体激动剂(咖啡因)和抑制剂(ryanodine)。在此之后,我们进行了一系列生物信息学分析，以发现参与认知功能障碍发展的蛋白质。Rt-PCR和Westernblot用于评估mRNA水平和蛋白质表达。结果：6h持续异氟醚麻醉可引起认知功能障碍，海马RyR3mRNA水平升高。麻醉后培养的N2a细胞中RyR3的mRNA水平与体内相当，RyR激动剂咖啡因纠正了麻醉后受到干扰的一些认知相关表型蛋白的表达。腹腔注射RyR激动剂咖啡因可改善异氟烷麻醉后小鼠的认知功能,生物信息学分析表明，CaMKⅣ可能参与了分子机制。
结论：Ryanodine受体激动剂咖啡因可改善异氟烷麻醉后小鼠的认知功能障碍。
结论：Ryanodine受体激动剂咖啡因可改善异氟烷麻醉后小鼠的认知功能障碍。
