Depolarization of mitochondrial membrane potential is the initial event in non-nucleoside reverse transcriptase inhibitor efavirenz induced cytotoxicity
Depolarization of mitochondrial membrane potential is the initial event in non-nucleoside reverse transcriptase inhibitor efavirenz induced cytotoxicity
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DOI：
10.1007/s10565-016-9362-9
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发表时间：
2017-02-01
影响因子：
6.1
通讯作者：
Chaubey, Binay
中科院分区：
文献类型：
--
作者：
Ganta, Krishna Kumar;Mandal, Anirban;Chaubey, Binay
依法韦仑是一种非核苷类逆转录酶抑制剂（NNRTI），是高效抗逆转录病毒治疗方案的活性成分。它对控制和管理人体免疫机能丧失病毒的传播作出了重大贡献。然而，EFV给药也导致了严重的不良反应，一些报告强调了EFV在线粒体功能障碍和毒性中的作用，但对其分子机制知之甚少。在本研究中，用临床相关浓度的EFV处理人肝癌细胞Huh 7.5，并评估诸如细胞毒性、线粒体跨膜电位、线粒体形态、细胞色素c释放、线粒体a介导的凋亡、mtDNA和mtDNA水平以及EFV分布到线粒体隔室中的参数，以了解导致EFV处理的细胞中细胞死亡的事件顺序。EFV在其临床相关浓度下在治疗48和72小时后具有显著毒性。EFV介导的毒性起始于线粒体外膜的透化和线粒体膜电位（Δ psi m）的变化，其触发一系列事件如细胞色素c释放、线粒体形态改变和线粒体介导的细胞凋亡。在IC50浓度下EFV处理48小时后，总线粒体含量降低，这也反映在线粒体DNA和RNA水平降低中。在与EFV孵育12小时后检测线粒体隔室中的EFV后，我们假设EFV是亲脂性分子，其被内化到线粒体隔室中，引起Δ psi m的去极化，其随后导致引起细胞死亡的级联事件。
Efavirenz is a non-nucleoside reverse transcriptase inhibitor (NNRTI) and an active constituent of the highly active antiretroviral therapy regime. It has significantly contributed in control and management of human immunodeficiency virus propagation. However, EFV administration has also led to severe adverse effects, several reports highlighted the role of EFV in mitochondrial dysfunction and toxicity but the molecular mechanism has been poorly understood. In present study, human hepatoma cells Huh 7.5 were treated with clinically relevant concentrations of EFV and parameters like cytotoxicity, mitochondrial transmembrane potential, mitochondrial morphology, cytochrome c release, mitochondria-mediated apoptosis, mtDNA and mtRNA levels and EFV distribution into mitochondrial compartment were evaluated to understand sequence of events leading to cell death in EFV-treated cells. EFV at its clinically relevant concentration was significantly toxic after 48 and 72 h of treatments. EFV-mediated toxicity is initiated with the permeabilization of mitochondrial outer membrane and change in mitochondrial membrane potential (Delta psi m) which triggers a series of events like cytochrome c release, alteration in mitochondrial morphology and mitochondria-mediated apoptosis. Total mitochondrial content is reduced after 48 h of EFV treatment at IC50 concentration which is also reflected in reduced mitochondrial DNA and RNA levels. After detecting EFV in mitochondrial compartment after 12 h of incubation with EFV, we hypothesize that EFV being a lipophilic molecule is internalized into the mitochondrial compartment causing depolarization of Delta psi m which subsequently leads to a cascade of events causing cell death.