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目的：通过体外实验探讨艾芬地尔对异氟烷所致发育期海马神经元毒性的保护作用。方法：从出生一天的大鼠海马获取神经元并体外培养5 天。这些神经元被随机分入4 组，包括对照组（control 组）、异氟醚组（Iso 组）、艾芬地尔（Ifen 组）和艾芬地尔+ 异氟烷组（Ifen + ISO 组）。使用MTT 法检测细胞活力及细胞损伤程度。使用TUNEL染色法检测细胞凋亡。使用Western blot 法检测神经元中NMDA 受体亚基NR2B和活化caspase-3 的表达水平。结果：与对照组比较，在2.4%异氟烷暴露后1 小时神经元的细胞活力显著下降（P<0.05）。同时，在2.4%异氟烷暴露后神经元的凋亡指数也显著升高（P<0.05）。Western blot 结果显示，异氟烷暴露可显著升高神经元活化caspase-3 和NR2B 的表达水平（P<0.05）。然而，使用NR2B拮抗剂艾芬地尔（20 uM）不仅可显著减少异氟烷所致的NR2B表达水平增高，也可缓解异氟烷造成的神经元凋亡和细胞损伤（P<0.01）。结论：异氟烷可导致发育期神经元NR2B 表达水平增高，而使用NR2B受体拮抗剂艾芬地尔可有效抑制NR2B的表达水平从而减少异氟烷所致神经元毒性。

英文摘要:

Objective:To investigate the neuroprotective effects of ifenprodil on isofluane-induced neurotoxicity in developing hippocampal neurons.Methods:Neurons harvested from hippocampi of postnatal day1 (P1) rats were cultured for 5 days and then randomly divided into control group (group control), isoflurane group (group ISO), ifenprodil group (group Ifen), ifenprodial + isoflurane group (group Ifen + ISO). MTT was used to detect cell viability. TUNEL assays were used to detect cell apoptosis. Western blot was conducted to detect protein expression of NR2B and active caspase-3.Results:Compared with control group, cell viability of cultured neurons significantly declined after 2.4% isoflurane exposure. The percentage of apoptotic cells in cultured neurons was significantly increased after 2.4%isoflurane exposure. Western blot analysis showed that isoflurane exposure significantly enhanced protein expressions of active caspase-3 and NR2B. However, the treatment of NR2B antagonist ifenprodil (20 uM) not only significantly reduced isoflurane- induced increased NR2B expressions, but also attenuated isoflurane-related neuronal apoptosis and cell injury.Conclusion:Our results suggest that ifenprodil treatments might be effective measurements for preventing isofluane-induced neurotoxicity in the developing hippocampal neurons.

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