Alvarez-Olmedo Daiana G, Biaggio Veronica S, Koumbadinga Geremy A, Gómez Nidia N, Shi Chunhua, Ciocca Daniel R, Batulan Zarah, Fanelli Mariel A, O'Brien Edward R
Oncology Laboratory at the Institute of Experimental Medicine and Biology of Cuyo (IMBECU), CCT CONICET Mendoza, Av. Ruiz Leal S/N Parque Gral. San Martin, CP 5500, Mendoza, CP, Argentina.
Libin Cardiovascular Institute of Alberta, Cumming School of Medicine, Foothills Medical Centre, University of Calgary, Room C823, 1403-29th Street NW, Calgary, AB, T2N 2T9, Canada.
Cell Stress Chaperones. 2017 May;22(3):357-369. doi: 10.1007/s12192-017-0768-y. Epub 2017 Mar 24.
Cadmium (Cd) is a carcinogen with several well-described toxicological effects in humans, but its molecular mechanisms are still not fully understood. Overexpression of heat shock protein 27 (HSP27/HSPB1)-a multifunctional protein chaperone-has been shown to protect cells from oxidative damage and apoptosis triggered by Cd exposure. The aims of this work were to investigate the potential use of extracellular recombinant HSP27 to prevent/counteract Cd-induced cellular toxicity and to evaluate if peroxynitrite was involved in the development of Cd-induced toxicity. Here, we report that the harmful effects of Cd correlated with changes in oxidative stress markers: upregulation of reactive oxygen species, reduction in nitric oxide (NO) bioavailability, increment in lipid peroxidation, peroxynitrite (PN), and protein nitration; intracellular HSP27 was reduced. Treatments with Cd (100 μM) for 24 h or with the peroxynitrite donor, SIN-1, decreased HSP27 levels (~50%), suggesting that PN formation is responsible for the reduction of HSP27. Pre-treatments of the cells either with Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME) (a pharmacological inhibitor of NO synthase) or with recombinant HSP27 (rHSP27) attenuated the disruption of the cellular metabolism induced by Cd, increasing in a 55 and 52%, respectively, the cell viability measured by CCK-8. Cd induced necrotic cell death pathways, although apoptosis was also activated; pre-treatment with L-NAME or rHSP27 mitigated cell death. Our findings show for the first time a direct relationship between Cd-induced toxicity and PN production and a role for rHSP27 as a potential therapeutic agent that may counteract Cd toxicity.
镉(Cd)是一种致癌物,对人体具有多种已被充分描述的毒理学效应,但其分子机制仍未完全明确。热休克蛋白27(HSP27/HSPB1)——一种多功能蛋白伴侣——的过表达已被证明可保护细胞免受镉暴露引发的氧化损伤和细胞凋亡。本研究的目的是探讨细胞外重组HSP27在预防/对抗镉诱导的细胞毒性方面的潜在用途,并评估过氧亚硝酸盐是否参与镉诱导毒性的发生发展。在此,我们报告镉的有害作用与氧化应激标志物的变化相关:活性氧上调、一氧化氮(NO)生物利用度降低、脂质过氧化、过氧亚硝酸盐(PN)和蛋白质硝化增加;细胞内HSP27减少。用100 μM镉处理24小时或用过氧亚硝酸盐供体SIN-1处理可使HSP27水平降低约50%,这表明PN的形成是HSP27减少的原因。用盐酸Nω-硝基-L-精氨酸甲酯(L-NAME)(一种NO合酶的药理学抑制剂)或重组HSP27(rHSP27)对细胞进行预处理,可减轻镉诱导的细胞代谢紊乱,分别使CCK-8检测的细胞活力提高55%和52%。镉诱导坏死性细胞死亡途径,尽管也激活了细胞凋亡;用L-NAME或rHSP27预处理可减轻细胞死亡。我们的研究结果首次表明镉诱导的毒性与PN产生之间存在直接关系,以及rHSP27作为一种可能对抗镉毒性的潜在治疗剂的作用。