Arend Christian, Rother Alica, Stolte Stefan, Dringen Ralf
Faculty 2 (Biology/Chemistry), Center for Biomolecular Interactions Bremen, University of Bremen, P.O. Box 330440, 28334, Bremen, Germany.
Center for Environmental Research and Sustainable Technology (UFT), University of Bremen, Bremen, Germany.
Neurochem Res. 2016 Dec;41(12):3278-3288. doi: 10.1007/s11064-016-2059-x. Epub 2016 Sep 21.
Efavirenz is a widely prescribed non-nucleoside reverse transcriptase inhibitor for the treatment of HIV infections. To test for potential long-term consequences of efavirenz on brain cells, cultured primary astrocytes were incubated with this substance or with its primary metabolite 8-hydroxy efavirenz for up to 7 days. Both, efavirenz and 8-hydroxy efavirenz caused time- and concentration-dependent cell toxicity and stimulated in subtoxic concentrations the glycolytic flux (glucose consumption and lactate release) in astrocytes. As 8-hydroxy efavirenz was less toxic than efavirenz and stimulated glycolysis in lower concentrations we tested for a potential hydroxylation of efavirenz to 8-hydroxy efavirenz in astrocytes. Analysis of media and cell lysates by HPLC-UV and mass spectrometry revealed that after 3 days of incubation viable astrocytes had accumulated about 17 and 7 % of the applied efavirenz and 8-hydroxy efavirenz, respectively. However, in cultures treated with efavirenz neither 8-hydroxy efavirenz nor any other known metabolite of efavirenz was detectable. These data demonstrate that cultured rat astrocytes efficiently accumulate, but not metabolize, efavirenz and 8-hydroxy efavirenz and that the observed chronic stimulation of glycolysis is mediated by both efavirenz and 8-hydroxy efavirenz.
依非韦伦是一种广泛用于治疗HIV感染的非核苷类逆转录酶抑制剂。为了测试依非韦伦对脑细胞的潜在长期影响，将培养的原代星形胶质细胞与该物质或其主要代谢产物8-羟基依非韦伦孵育长达7天。依非韦伦和8-羟基依非韦伦均引起时间和浓度依赖性细胞毒性，并在亚毒性浓度下刺激星形胶质细胞中的糖酵解通量（葡萄糖消耗和乳酸释放）。由于8-羟基依非韦伦比依非韦伦毒性小且在较低浓度下刺激糖酵解，我们测试了星形胶质细胞中依非韦伦是否可能羟基化为8-羟基依非韦伦。通过HPLC-UV和质谱对培养基和细胞裂解物进行分析，结果显示，孵育3天后，存活的星形胶质细胞分别积累了约17%和7%的所施加的依非韦伦和8-羟基依非韦伦。然而，在用依非韦伦处理的培养物中，未检测到8-羟基依非韦伦或依非韦伦的任何其他已知代谢产物。这些数据表明，培养的大鼠星形胶质细胞能有效积累但不能代谢依非韦伦和8-羟基依非韦伦，并且观察到的糖酵解慢性刺激是由依非韦伦和8-羟基依非韦伦共同介导的。
