Aβ_(25～35)对海马神经元的损伤及双苯氟嗪的保护作用

Injury of hippocampal neuron induced by Aβ_(25～35) and protective effect of dipfluzine

目的观察Aβ25～35对原代培养的胎鼠海马神经元的损伤作用以及双苯氟嗪的保护作用。方法将老化的Aβ25～35加入到培养7d的海马神经细胞中,采用乳酸脱氢酶试剂盒测定海马神经元乳酸脱氢酶释放度(LDH%),观察神经元的损伤情况。选择1.0μmol/LAβ25～35制备海马神经元损伤模型,乳酸脱氢酶试剂盒测定海马神经元LDH%,激光扫描共聚焦显微镜测定钙荧光强度,观察双苯氟嗪对神经元损伤的保护作用。结果Aβ25～350.1、1.0及10.0μmol/L均能明显升高海马神经元LDH%,造成神经元损伤。给予1.0和10.0μmol/L双苯氟嗪均显著降低1.0μmol/LAβ25～35升高的LDH%,与空白对照组相比,模型组细胞内钙荧光强度显著升高,而双苯氟嗪(1.0和10.0μmol/L)处理后明显降低。结论双苯氟嗪对Aβ25～35造成的海马神经元损伤有保护作用,其机制可能与抑制钙超载有关。

Objective To study the injury effect of Aβ25～35 on primary cultured hippocampal neuron of foetal rats and the protective effect of dipfluzine. Methods Aβ25～35 was added to the hippocampal neurons which had been cultured for 7 d, then the injury effect was evaluated by the lactate dehydrogenase release （ LDH% ） 24 h afterAβ25～35 was added. Aβ25～35 1.0 μmol/L was chosen for model preparation. LDH% was assayed by LDH kit. The level of intracellular calcium fluorescence intensity was assayed by laser scanning confocal microscope. Results The LDH% was increased obviously by Aβ25～35 （0. 1, 1.0 and 10. 0 μmol/L）. The LDH% elevated by Aβ25～35 1.0 μmol/L was decreased by treatment with dipfluzine （ 1.0 and 10. 0 μmoL/L）. The level of intracellular calcium fluorescence intensity in the group treated with Aβ25～35 was far higher than that in blank group and groups treated with dipfluzine （ 1.0 and 10.0 μmol/L）. Conclusions Dipfluzine has protective effect against injury of hippocampal neurons induced by Aβ25～35, which is related to decreasing concentration of intracellular free calcium.