Yan Ming, Zhang Yun, Qin Haiyan, Liu Kezhou, Guo Miao, Ge Yakun, Xu Mingen, Sun Yonghong, Zheng Xiaoxiang
Department of Biomedical Engineering, College of Life Information Science and Instrument Engineering, Hangzhou Dianzi University, Hangzhou, People's Republic of China.
Basic Medical Sciences, College of Medicine, Shaoxing University, Shaoxing, People's Republic of China.
Int J Nanomedicine. 2016 Feb 2;11:529-42. doi: 10.2147/IJN.S93591. eCollection 2016.
Cadmium telluride quantum dots (CdTe QDs) have been proposed to induce oxidative stress, which plays a crucial role in CdTe QDs-mediated mitochondrial-dependent apoptosis in human umbilical vein endothelial cells (HUVECs). However, the direct interactions of CdTe QDs with HUVECs and their potential impairment of other organelles like endoplasmic reticulum (ER) in HUVECs are poorly understood. In this study, we reported that the negatively charged CdTe QDs (-21.63±0.91 mV), with good dispersity and fluorescence stability, were rapidly internalized via endocytosis by HUVECs, as the notable internalization could be inhibited up to 95.52% by energy depletion (NaN3/deoxyglucose or low temperature). The endocytosis inhibitors (methyl-β-cyclodextrin, genistein, sucrose, chlorpromazine, and colchicine) dramatically decreased the uptake of CdTe QDs by HUVECs, suggesting that both caveolae/raft- and clathrin-mediated endocytosis were involved in the endothelial uptake of CdTe QDs. Using immunocytochemistry, a striking overlap of the internalized CdTe QDs and ER marker was observed, which indicates that QDs may be transported to ER. The CdTe QDs also caused remarkable ER stress responses in HUVECs, confirmed by significant dilatation of ER cisternae, upregulation of ER stress markers GRP78/GRP94, and activation of protein kinase RNA-like ER kinase-eIF2α-activating transcription factor 4 pathway (including phosphorylation of both protein kinase RNA-like ER kinase and eIF2α and elevated level of activating transcription factor 4). CdTe QDs further promoted an increased C/EBP homologous protein expression, phosphorylation of c-JUN NH2-terminal kinase, and cleavage of ER-resident caspase-4, while the specific inhibitor (SP600125, Z-LEVD-fmk, or salubrinal) significantly attenuated QDs-triggered apoptosis, indicating that all three ER stress-mediated apoptosis pathways were activated and the direct participation of ER in the CdTe QDs-caused apoptotic cell death in HUVECs. Our findings provide important new insights into QDs toxicity and reveal potential cardiovascular risks for the future applications of QDs.
碲化镉量子点(CdTe QDs)已被提出可诱导氧化应激,氧化应激在CdTe QDs介导的人脐静脉内皮细胞(HUVECs)线粒体依赖性凋亡中起关键作用。然而,CdTe QDs与HUVECs的直接相互作用及其对HUVECs中内质网(ER)等其他细胞器的潜在损害尚不清楚。在本研究中,我们报道带负电荷的CdTe QDs(-21.63±0.91 mV)具有良好的分散性和荧光稳定性,通过HUVECs的内吞作用迅速内化,因为能量耗竭(NaN3/脱氧葡萄糖或低温)可将显著的内化作用抑制高达95.52%。内吞抑制剂(甲基-β-环糊精、染料木黄酮、蔗糖、氯丙嗪和秋水仙碱)显著降低了HUVECs对CdTe QDs的摄取,表明小窝/脂筏介导的内吞作用和网格蛋白介导的内吞作用均参与了内皮细胞对CdTe QDs的摄取。通过免疫细胞化学观察到内化的CdTe QDs与ER标志物有明显重叠,这表明量子点可能被转运至内质网。CdTe QDs还在HUVECs中引起显著的内质网应激反应,表现为内质网池显著扩张、内质网应激标志物GRP78/GRP94上调以及蛋白激酶RNA样内质网激酶-eIF2α-激活转录因子4通路的激活(包括蛋白激酶RNA样内质网激酶和eIF2α的磷酸化以及激活转录因子4水平升高)。CdTe QDs进一步促进了C/EBP同源蛋白表达增加、c-JUN NH2末端激酶磷酸化以及内质网驻留半胱天冬酶-4的切割,而特异性抑制剂(SP600125、Z-LEVD-fmk或沙芦比诺)显著减轻了量子点触发的凋亡,表明内质网应激介导的三条凋亡途径均被激活,且内质网直接参与了CdTe QDs导致的HUVECs凋亡性细胞死亡。我们的研究结果为量子点毒性提供了重要的新见解,并揭示了量子点未来应用中的潜在心血管风险。