Feng Yan, Wang Kui, Wang Ning, Jia Pengyu, Zhang Lei, Yuan Haozheng, Lu Pan, Lu Yang, Zhang Hong, Li Rong, Zhang Yan, Li Qianqian, Zhang Pengbo
Department of Anesthesiology, The Second Affiliated Hospital of Xi'an Jiaotong University, 157# West 5 Road, 710004, Xi'an, Shaanxi, China.
Department of Anesthesiology, Xi'an People's Hospital (Xi'an Fourth Hospital), 710004, Xi'an, Shaanxi, China.
Metab Brain Dis. 2022 Oct;37(7):2457-2466. doi: 10.1007/s11011-022-01008-2. Epub 2022 Jul 15.
Sevoflurane, a commonly used anesthetic, has been found to cause neural stem cell (NSC) injury, thereby contributing to neurocognitive impairment following general anesthesia. Tetramethylpyrazine (TMP), one of the most widely used medicinal compounds isolated from a traditional Chinese herb, possess neuroprotective activity. However, its effect on sevoflurane-induced NSC injury remains unclear. NSCs were pretreated with indicated concentrations of TMP for 2 h and then exposed to sevoflurane for 6 h. Cell injury was measured using lactate dehydrogenase (LDH) release assay. Cell viability and proliferation were detected by cell counting kit-8 (CCK-8) assay and 5-bromo-2'-deoxyuridine (BrdU) labeling, respectively. Apoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The levels of cleaved caspase-3, phosphorylated protein kinase B (Akt) and phosphorylated glycogen synthase kinase-3β (GSK-3β) were detected by western blotting. Our results showed exposure to sevoflurane decreased the viability and proliferation of NSCs, while TMP preserved NSC viability and proliferation after sevoflurane exposure. In addition, the expression of cleaved caspase-3 and TUNEL positive cells were markedly decreased in TMP-treated NSCs compared with the control. Furthermore, pretreatment with TMP significantly increased the levels of phosphorylated Akt and GSK-3β in sevoflurane-injured NSCs. However, an upstream inhibitor of Akt, LY294002 abolished the protective of TMP on the cell viability of NSCs. In conclusion, these findings indicate that TMP protects NSCs from sevoflurane-induced toxicity through Akt/GSK-3β pathway.
七氟醚是一种常用的麻醉剂，已被发现会导致神经干细胞（NSC）损伤，从而导致全身麻醉后出现神经认知障碍。川芎嗪（TMP）是从一种传统中药中分离出的使用最广泛的药用化合物之一，具有神经保护活性。然而，其对七氟醚诱导的神经干细胞损伤的影响仍不清楚。将神经干细胞用指定浓度的川芎嗪预处理2小时，然后暴露于七氟醚中6小时。使用乳酸脱氢酶（LDH）释放试验测量细胞损伤。分别通过细胞计数试剂盒-8（CCK-8）试验和5-溴-2'-脱氧尿苷（BrdU）标记检测细胞活力和增殖。使用末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）试验检测凋亡细胞。通过蛋白质免疫印迹法检测裂解的半胱天冬酶-3、磷酸化蛋白激酶B（Akt）和磷酸化糖原合酶激酶-3β（GSK-3β）的水平。我们的结果表明，暴露于七氟醚会降低神经干细胞的活力和增殖，而川芎嗪在七氟醚暴露后可维持神经干细胞的活力和增殖。此外与对照组相比，经川芎嗪处理的神经干细胞中裂解的半胱天冬酶-3的表达和TUNEL阳性细胞明显减少。此外，用川芎嗪预处理可显著提高七氟醚损伤的神经干细胞中磷酸化Akt和GSK-3β的水平。然而，Akt的上游抑制剂LY294002消除了川芎嗪对神经干细胞细胞活力的保护作用。总之，这些发现表明川芎嗪通过Akt/GSK-3β途径保护神经干细胞免受七氟醚诱导的毒性。
