Uesono Yukifumi, Toh-e Akio, Kikuchi Yoshiko, Araki Tomoyuki, Hachiya Takushi, Watanabe Chihiro K, Noguchi Ko, Terashima Ichiro
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 113-0033 Japan
Medical Mycology Research Center (MMRC), Chiba University, Chiba, 260-8673 Japan.
Genetics. 2016 Mar;202(3):997-1012. doi: 10.1534/genetics.115.183806. Epub 2016 Jan 12.
Action mechanisms of anesthetics remain unclear because of difficulty in explaining how structurally different anesthetics cause similar effects. In Saccharomyces cerevisiae, local anesthetics and antipsychotic phenothiazines induced responses similar to those caused by glucose starvation, and they eventually inhibited cell growth. These drugs inhibited glucose uptake, but additional glucose conferred resistance to their effects; hence, the primary action of the drugs is to cause glucose starvation. In hxt(0) strains with all hexose transporter (HXT) genes deleted, a strain harboring a single copy of HXT1 (HXT1s) was more sensitive to tetracaine than a strain harboring multiple copies (HXT1m), which indicates that quantitative reduction of HXT1 increases tetracaine sensitivity. However, additional glucose rather than the overexpression of HXT1/2 conferred tetracaine resistance to wild-type yeast; therefore, Hxts that actively transport hexoses apparently confer tetracaine resistance. Additional glucose alleviated sensitivity to local anesthetics and phenothiazines in the HXT1m strain but not the HXT1s strain; thus, the glucose-induced effects required a certain amount of Hxt1. At low concentrations, fluorescent phenothiazines were distributed in various membranes. At higher concentrations, they destroyed the membranes and thereby delocalized Hxt1-GFP from the plasma membrane, similar to local anesthetics. These results suggest that the aforementioned drugs affect various membrane targets via nonspecific interactions with membranes. However, the drugs preferentially inhibit the function of abundant Hxts, resulting in glucose starvation. When Hxts are scarce, this preference is lost, thereby mitigating the alleviation by additional glucose. These results provide a mechanism that explains how different compounds induce similar effects based on lipid theory.
由于难以解释结构不同的麻醉剂如何产生相似的效果,麻醉剂的作用机制仍不清楚。在酿酒酵母中,局部麻醉剂和抗精神病吩噻嗪类药物诱导的反应类似于葡萄糖饥饿所引起的反应,最终抑制细胞生长。这些药物抑制葡萄糖摄取,但额外添加葡萄糖可使其产生抗性;因此,这些药物的主要作用是导致葡萄糖饥饿。在所有己糖转运蛋白(HXT)基因均被删除的hxt(0)菌株中,携带单个HXT1拷贝(HXT1s)的菌株比携带多个拷贝(HXT1m)的菌株对丁卡因更敏感,这表明HXT1的定量减少会增加丁卡因敏感性。然而,额外添加葡萄糖而非HXT1/2的过表达赋予野生型酵母丁卡因抗性;因此,主动转运己糖的Hxts显然赋予丁卡因抗性。额外添加葡萄糖减轻了HXT1m菌株对局部麻醉剂和吩噻嗪类药物的敏感性,但对HXT1s菌株无效;因此,葡萄糖诱导的效应需要一定量的Hxt1。在低浓度下,荧光吩噻嗪类药物分布在各种膜中。在较高浓度下,它们破坏膜,从而使Hxt1-GFP从质膜上脱离,类似于局部麻醉剂。这些结果表明,上述药物通过与膜的非特异性相互作用影响各种膜靶点。然而,这些药物优先抑制丰富的Hxts的功能,导致葡萄糖饥饿。当Hxts稀缺时,这种偏好就会丧失,从而减轻额外添加葡萄糖所带来的缓解作用。这些结果提供了一种基于脂质理论解释不同化合物如何诱导相似效应的机制。