布美他尼消除两性霉素B诱导的细胞凋亡和细胞毒性是由于其对细胞钾离子通量的影响。

Bumetanide annihilation of amphotericin B-induced apoptosis and cytotoxicity is due to its effect on cellular K+ flux.

作者信息

Marklund L, Behnam-Motlagh P, Henriksson R, Grankvist K

机构信息

Department of Clinical Chemistry, Umeå University, S-901 87 Umeå, Sweden.

出版信息

J Antimicrob Chemother. 2001 Dec;48(6):781-6. doi: 10.1093/jac/48.6.781.

DOI:10.1093/jac/48.6.781

PMID:11733461

Abstract

The antifungal antibiotic amphotericin B causes considerable toxic effects during clinical therapy. We have shown previously that amphotericin B-induced cytotoxicity and apoptosis were eradicated by the Na+, K+, 2Cl- cotransport inhibitor bumetanide. To elucidate the role of K+ flux and the activity of Na+, K+ ATPase and Na+, K+, 2Cl- cotransport in apoptosis and cytotoxicity induced by amphotericin B alone and combined with bumetanide, we quantified the influx and efflux of K+ of mesothelioma cells (P31) using the K+ analogue 86Rb+ with ouabain (100 micromol/L) as the K+ influx probe. To determine the susceptibility of Candida albicans to amphotericin B when combined with bumetanide we used a plate diffusion method. Amphotericin B or bumetanide alone significantly stimulated 86Rb+ efflux during the first 15 min. However, when added simultaneously, the cellular 86Rb+ efflux was markedly decreased. Amphotericin B (3 mg/L) had no effect on immediate (15 min) total 86Rb+ influx. When bumetanide (100 micromol/L) was added, the total 86Rb+ influx was markedly reduced due to inhibition of augmented Na+, K+, 2Cl- cotransport and low Na+, K+ ATPase activity. Bumetanide did not affect the susceptibility of C. albicans to amphotericin B, which suggests that bumetanide or related drugs could be used in antifungal therapy to increase amphotericin B effectiveness without increasing its adverse effects. We suggest that bumetanide hampering of amphotericin B-induced cytotoxicity and apoptosis could be due to an immediate reduction of cellular K+ efflux as well as disordered K+ influx.

摘要

抗真菌抗生素两性霉素B在临床治疗期间会引起相当大的毒性作用。我们之前已经表明，两性霉素B诱导的细胞毒性和凋亡可被Na⁺、K⁺、2Cl⁻共转运抑制剂布美他尼消除。为了阐明K⁺通量以及Na⁺、K⁺ ATP酶和Na⁺、K⁺、2Cl⁻共转运的活性在两性霉素B单独诱导以及与布美他尼联合诱导的凋亡和细胞毒性中的作用，我们使用K⁺类似物⁸⁶Rb⁺和哇巴因（100 μmol/L）作为K⁺内流探针，对间皮瘤细胞（P31）的K⁺流入和流出进行了定量。为了确定白色念珠菌与布美他尼联合使用时对两性霉素B的敏感性，我们采用了平板扩散法。单独使用两性霉素B或布美他尼在最初15分钟内显著刺激了⁸⁶Rb⁺外流。然而，当同时添加时，细胞的⁸⁶Rb⁺外流明显减少。两性霉素B（3 mg/L）对即刻（15分钟）的总⁸⁶Rb⁺内流没有影响。当添加布美他尼（100 μmol/L）时，由于增强的Na⁺、K⁺、2Cl⁻共转运受到抑制以及Na⁺、K⁺ ATP酶活性降低，总⁸⁶Rb⁺内流明显减少。布美他尼不影响白色念珠菌对两性霉素B的敏感性，这表明布美他尼或相关药物可用于抗真菌治疗，以提高两性霉素B的疗效而不增加其不良反应。我们认为布美他尼抑制两性霉素B诱导的细胞毒性和凋亡可能是由于细胞K⁺外流立即减少以及K⁺内流紊乱。