度洛西汀对小脑颗粒神经元谷氨酸毒性作用的保护作用及机制(1)
[摘要] 目的 观察度洛西汀对小脑颗粒神经元谷氨酸毒性作用的保护作用及机制。 方法 体外培养CD-1小鼠小脑颗粒神经元，神经元细胞随机分为8组：对照组，谷氨酸组，度洛西汀组，度洛西汀+谷氨酸组，SB204741处理组，SB204741+谷氨酸组，LY294002处理组，LY294002+谷氨酸组。采用TUNEL法检测神经元细胞凋亡程度，采用共聚焦显微镜fura-2探针检测神经元内Ca2+水平。 结果 与对照组比较，谷氨酸毒性能明显增加神经元凋亡[(1.62±0.96)比(13.98±3.49)](P < 0.05)，度洛西汀顯著抑制由谷氨酸毒性引起的神经元凋亡。度洛西汀能降低由谷氨酸毒性激活的神经元内Ca2+升高。5-HT2B SB204741能抑制由谷氨酸毒性引起的神经元凋亡和神经元内Ca2+升高，而LY294002只能抑制谷氨酸毒性引起的神经元凋亡，对于谷氨酸毒性引起的神经元内Ca2+升高无作用。 结论 度洛西汀具有抵抗神经元谷氨酸毒性作用，其机制主要是通过5-HT2B受体活化细胞内钙库释放，从而引起EGFR间接激活PI3K/AKT/mTOR通路，抑制由谷氨酸毒性引起的神经元凋亡。
[关键词] 度洛西汀；谷氨酸毒性；神经元
[中图分类号] R971 [文献标识码] A [文章编号] 1673-7210(2016)11(c)-0030-04
Protective effect of Duloxetine on glutamate toxicity of cerebellar granule neurons and its mechanism
LIU Sixin HE Dan YU Xiaojun WANG Jiaqi
The Second Department of Neurology， the First Hospital of Changsha City， Hu'nan Province， Changsha 410005， China
[Abstract] Objective To observe the protective effect of Duloxetine on glutamate toxicity of cerebellar granule neurons and its mechanism. Methods The cerebellar granule neurons derived from CD-1 mice were cultured in vitro， all the neurons were randomly divided into eight groups： control group， glutamate group， Duloxetine group， Duloxetine+glutamate group， SB204741 treatment group， SB204741+glutamate group， LY294002 treatment group， LY294002+glutamate group. The apoptosis degrees of neurons were detected by TUNEL method， the levels of Ca2+ in neurons were detected by confocal microscopy fura-2. Results Compared with control group， glutamate toxicity could significantly increase the neuron apoptosis [(1.62±0.96) vs (13.98±3.49)] (P < 0.05)， and Duloxetine could inhibit the neuron apoptosis caused by glutamate toxicity. Duloxetine could significantly decrease the increasing concentration of Ca2+ in neurons activated by glutamate toxicity. 5-HT2B SB204741 could significantly inhibit the neuron apoptosis and the increasing concentration of Ca2+ in neurons caused by glutamate toxicity， while LY294002 could only inhibit the neuron apoptosis caused by glutamate toxicity， which had no effects on the increasing concentration of Ca2+ in neurons activated by glutamate toxicity. Conclusion Duloxetine has the effects of inhibiting the glutamate toxicity of neurons， the main mechanism may be associated with the release of intracellular calcium store activated by the receptor of 5-HT2B， which cause EGFR activating PI3K/AKT/mTOR signal pathway indirectly， so as to inhibit the neuron apoptosis caused by glutamate toxicity., 百拇医药(柳四新 何丹 余孝君 王家祺)
[关键词] 度洛西汀；谷氨酸毒性；神经元
[中图分类号] R971 [文献标识码] A [文章编号] 1673-7210(2016)11(c)-0030-04
Protective effect of Duloxetine on glutamate toxicity of cerebellar granule neurons and its mechanism
LIU Sixin HE Dan YU Xiaojun WANG Jiaqi
The Second Department of Neurology， the First Hospital of Changsha City， Hu'nan Province， Changsha 410005， China
[Abstract] Objective To observe the protective effect of Duloxetine on glutamate toxicity of cerebellar granule neurons and its mechanism. Methods The cerebellar granule neurons derived from CD-1 mice were cultured in vitro， all the neurons were randomly divided into eight groups： control group， glutamate group， Duloxetine group， Duloxetine+glutamate group， SB204741 treatment group， SB204741+glutamate group， LY294002 treatment group， LY294002+glutamate group. The apoptosis degrees of neurons were detected by TUNEL method， the levels of Ca2+ in neurons were detected by confocal microscopy fura-2. Results Compared with control group， glutamate toxicity could significantly increase the neuron apoptosis [(1.62±0.96) vs (13.98±3.49)] (P < 0.05)， and Duloxetine could inhibit the neuron apoptosis caused by glutamate toxicity. Duloxetine could significantly decrease the increasing concentration of Ca2+ in neurons activated by glutamate toxicity. 5-HT2B SB204741 could significantly inhibit the neuron apoptosis and the increasing concentration of Ca2+ in neurons caused by glutamate toxicity， while LY294002 could only inhibit the neuron apoptosis caused by glutamate toxicity， which had no effects on the increasing concentration of Ca2+ in neurons activated by glutamate toxicity. Conclusion Duloxetine has the effects of inhibiting the glutamate toxicity of neurons， the main mechanism may be associated with the release of intracellular calcium store activated by the receptor of 5-HT2B， which cause EGFR activating PI3K/AKT/mTOR signal pathway indirectly， so as to inhibit the neuron apoptosis caused by glutamate toxicity., 百拇医药(柳四新 何丹 余孝君 王家祺)