Gola Joanna, Strzałka-Mrozik Barbara, Kruszniewska-Rajs Celina, Janiszewski Adrian, Skowronek Bartłomiej, Gagoś Mariusz, Czernel Grzegorz, Mazurek Urszula
Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
Pharmacol Rep. 2017 Feb;69(1):22-28. doi: 10.1016/j.pharep.2016.09.008. Epub 2016 Sep 5.
A new form of amphotericin B (AmB)- complex with copper (II) ions (AmB-Cu) - is less toxic to human renal cells. Cytokines, including Tumor Necrosis Factor (TNF), are responsible for nephrotoxicity observed in patients treated with AmB. Another problem during therapy is the occurrence of oxidized forms of AmB (AmB-ox) in patients' circulation. To elucidate the molecular mechanism responsible for the reduction of the toxicity of AmB-Cu, we evaluated the expression of genes encoding TNF and its receptors alongside encoding proteins involved in TNF-induced signalization.
Renal cells (RPTECs) were treated with AmB, AmB-Cu or AmB-ox. The expression of TNF and its receptors was evaluated by ELISA tests and real-time RT-qPCR. The expression of TNF-related genes was appointed using oligonucleotide microarrays.
Only sTNFR1 was detected, and its level was lower in AmB-Cu- and AmB-ox-treated cells. TNFR1 mRNA was downregulated in AmB-ox, while TNFR2 mRNA was upregulated in AmB and AmB-Cu. Several changes in the expression of TNF-related genes coincided with changes in the expression of TNF receptors.
The lower toxicity of AmB-Cu could result from the changes in the expression of TNF receptors, which coincided with the changes in the expression of genes encoding proteins involved in TNF-induced pathways. This situation might subsequently result in a changes in intracellular signalization and influence the toxicity of tested forms of AmB on renal cells.
一种新的两性霉素B（AmB）与铜（II）离子的复合物（AmB-Cu）对人肾细胞的毒性较小。细胞因子，包括肿瘤坏死因子（TNF），是接受AmB治疗的患者中观察到的肾毒性的原因。治疗期间的另一个问题是患者循环中出现AmB的氧化形式（AmB-ox）。为了阐明负责降低AmB-Cu毒性的分子机制，我们评估了编码TNF及其受体以及参与TNF诱导信号传导的蛋白质的基因的表达。
用AmB、AmB-Cu或AmB-ox处理肾细胞（RPTECs）。通过ELISA试验和实时RT-qPCR评估TNF及其受体的表达。使用寡核苷酸微阵列确定TNF相关基因的表达。
仅检测到sTNFR1，其在AmB-Cu和AmB-ox处理的细胞中的水平较低。TNFR1 mRNA在AmB-ox中下调，而TNFR2 mRNA在AmB和AmB-Cu中上调。TNF相关基因表达的几种变化与TNF受体表达的变化一致。
AmB-Cu较低的毒性可能源于TNF受体表达的变化，这与编码参与TNF诱导途径的蛋白质的基因表达的变化一致。这种情况随后可能导致细胞内信号传导的变化，并影响所测试的AmB形式对肾细胞的毒性。
