TEA敏感型钾电流在gp120引起的海马神经元损伤中的作用
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国家自然科学基金面上项目(81072329);国家自然科学基金青年基金(81202230);南京医科大学科技发展基金(2011NJMU275);江苏省高校优势学科建设工程资助项目(CX09B-264Z);临床医学科技专项(SBL2014020070)


Involvements of TEA sensitive K+ currents in gp120 induced hippocampal neuronal apoptosis
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    摘要:

    目的:探讨gp120对TEA敏感型钾电流的作用以及该电流在gp120引起神经元损伤中的作用。方法:分离怀孕18 d SD大鼠胎鼠的海马神经元作为实验对象,分为对照组和gp120处理组,利用全细胞膜片钳的方法记录外向延迟钾电流的变化,通过MTT和TUNEL的方法分别观察gp120处理后神经元的活力和凋亡情况。Western blot观察Kv2.1钾通道的蛋白表达变化以及在TEA敏感型钾电流中的贡献度。结果:gp120能引起外向钾电流明显增大,具有浓度依赖性,其中TEA敏感型成分参与了外向钾电流的增大。此外,MTT和TUNEL实验发现,gp120能引起神经元活力的下降和凋亡,而钾通道抑制剂TEA能缓解由gp120引起的细胞损伤。进一步实验表明,gp120可上调TEA敏感型成分中Kv2.1钾通道的表达,且其特异性抑制剂GxTX-1E能显著抑制神经元中TEA敏感型钾电流成分。结论:TEA敏感型钾电流参与了gp120引起的神经元损伤过程,其重要组成成分Kv2.1通道蛋白上调,提示该通道蛋白可能参与了gp120引起的细胞损伤。

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    Objective:To explore the effects of gp120 on TEA sensitive K+ currents and the role of the currents in gp120 induced neuronal apoptosis. Methods: Hippocampal neurons were harvest from 18-day-old embryonic rats and divided into two groups: the control and the gp120 treated group. The outward delayed K+ currents were recorded by performing the whole cell patch clamp, subsequently the cell viability as well as neuron apoptosis were evaluated by MTT and TUNEL assay,respectively. Moreover, the protein expression of Kv2.1, a subtype of TEA sensitive K+ channel, was detected by Western blot. Results: Gp120 significantly increased the outward K+ currents in a dose-dependent manner, and the TEA sensitive K+ currents were participated in K+ currents increment mediated by gp120. The MTT and TUNEL results revealed that gp120 substantially decreased the cell viability and enhanced the cell apoptosis, while the K+ antagonist TEA was effectively decreased gp120 induced cell damage. Moreover, the expression and contribution of Kv2.1 on TEA sensitive K+ currents was significantly up-regulated by gp120. And the specific blocker, GxTX-1E, was used for evaluating the role of Kv2.1 in TEA sensitive K+ currents. Conclusion: TEA sensitive K+ currents were involved in gp120 induced neuronal damage, and the subtype Kv2.1, which was up-regulated by gp120, might exert effects on gp120 induced cell damage.

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余鹏飞,蒋 雷,钱文溢,周 景,王易欣,肖 杭,王 军. TEA敏感型钾电流在gp120引起的海马神经元损伤中的作用[J].南京医科大学学报(自然科学版),2015,(1):11-16

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  • 收稿日期:2014-06-09
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  • 在线发布日期: 2015-02-06
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