Julia V Bukanova, Rodion V Kondratenko, Elena I Solntseva
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引用次数: 0
摘要
异孕酮（Allo）是GABAA受体的正变构调节剂，阿米洛利（Ami）是GABAA受体的竞争性拮抗剂。本研究旨在研究Allo和Ami对GABAA受体功能活性的联合影响。采用膜片钳技术和快速应用系统对大鼠小脑浦肯野细胞gaba诱导的氯化电流（IGABA）进行了测定。结果表明，Allo抑制了Ami对IGABA的抑制作用，在Allo （EC50 = 1.2µM）、Ami （EC50 = 25.5µM）和Allo+Ami （EC50 = 3.2µM）存在的情况下，Ami浓度响应曲线的IC50值从164µM增加到547µM （p50 = 5.8µM）。计算EC50值相对于对照组的百分比变化。Allo存在时Ami的阻断作用降低（340% vs 150%, P < 0.05）。结果表明，GABAA受体上的Allo和Ami结合位点之间存在一种单向的变构关系，即从Allo位点到Ami位点，而不是相反。
Interaction Between Allopregnanolone and Amiloride Binding Sites on the GABAA Receptor.
Allopregnanolone (Allo) is a positive allosteric modulator of the GABAA receptor, and amiloride (Ami) is a competitive antagonist of the GABAA receptor. The purpose of this work was to study the combined effect of Allo and Ami on functional activity of GABAA receptor. The GABA-induced chloride current (IGABA) was measured in isolated Purkinje cells of rat cerebellum using the patch-clamp technique and a system of fast application. Our results indicate that Allo suppresses the inhibitory effect of Ami on IGABA, the IC50 value of Ami concentration-response curve was increased from 164 to 547 µM (P < 0.001) in the presence of Allo. Next, GABA concentration-response curves (EC50 = 5.8 µM) were constructed in the presence of Allo (EC50 = 1.2 µM), Ami (EC50 = 25.5 µM), and the combination of Allo+Ami (EC50 = 3.2 µM). Changes in EC50 values as a percentage relative to the control were calculated. The blocking effect of Ami is reduced in the presence of Allo (340% vs 150%, P < 0.01) and the potentiating effect of Allo does not change in the presence of Ami (78% vs 87%, P > 0.05). The results suggest that there is an allosteric relationship between the Allo and Ami binding sites on GABAA receptor that operates in one direction, from Allo sites to Ami site, but not vice versa.
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