Nanoparticles with High-Surface Negative-Charge Density Disturb the Metabolism of Low-Density Lipoprotein in Cells
Abstract
:1. Introduction
2. Results
2.1. Characterization of AuNPs
2.2. Effects of AuNPs on LDL Uptake in Cells
2.3. Effects of AuNPs on LDL Binding to LDL-R on the Cell Surface
2.4. Effects of AuNPs on LDL-R Gene Expression
2.5. Effects of AuNPs on LDL Transport in Cells
2.6. Retained LDL-R Was Degraded in Lysosomes
2.7. AuNPs Disrupted pH Regulation in Endosomes
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatment
4.2. AuNPs Synthesis
4.3. AuNPs Characterization
4.4. LDL Uptake Assay
4.5. LDL-R on Cell Surface Assay
4.6. Western Blot Assay
4.7. Reverse-Transcription PCR and Quantitative Real-Time PCR Analysis
4.8. Immunofluorescence Assay
4.9. Endocytic pH Measurement
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Brown, M.S.; Goldstein, J.L. Receptor-mediated endocytosis: Insights from the lipoprotein receptor system. Proc. Natl. Acad. Sci. USA 1979, 76, 3330–3337. [Google Scholar] [CrossRef] [PubMed]
- Zhang, S.; Gao, H.; Bao, G. Physical principles of nanoparticle cellular endocytosis. ACS Nano 2015, 9, 8655–8671. [Google Scholar] [CrossRef] [PubMed]
- Canton, I.; Battaglia, G. Endocytosis at the nanoscale. Chem. Soc. Rev. 2012, 41, 2718–2739. [Google Scholar] [CrossRef] [PubMed]
- Anderson, R.G.; Brown, M.S.; Goldstein, J.L. Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts. Cell 1977, 10, 351–364. [Google Scholar] [CrossRef]
- Goldstein, J.L.; Anderson, R.G.W.; Brown, M.S. Coated pits, coated vesicles, and receptor-mediated endocytosis. Nature 1979, 279, 679. [Google Scholar] [CrossRef] [PubMed]
- Vasile, E.; Simionescu, M.; Simionescu, N. Visualization of the binding, endocytosis, and transcytosis of low-density lipoprotein in the arterial endothelium in situ. J. Cell Biol. 1983, 96, 1677–1689. [Google Scholar] [CrossRef] [PubMed]
- Johnson, L.S.; Dunn, K.W.; Pytowski, B.; McGraw, T.E. Endosome acidification and receptor trafficking: Bafilomycin A1 slows receptor externalization by a mechanism involving the receptor’s internalization motif. Mol. Biol. Cell 1993, 4, 1251–1266. [Google Scholar] [CrossRef] [PubMed]
- Goldstein, J.L.; Brown, M.S. A century of cholesterol and coronaries: From plaques to genes to statins. Cell 2015, 161, 161–172. [Google Scholar] [CrossRef] [PubMed]
- Ghosh, P.S.; Kim, C.K.; Han, G.; Forbes, N.S.; Rotello, V.M. Efficient gene delivery vectors by tuning the surface charge density of amino acid-functionalized gold nanoparticles. ACS Nano 2008, 2, 2213–2218. [Google Scholar] [CrossRef] [PubMed]
- Kim, H.R.; Gil, S.; Andrieux, K.; Nicolas, V.; Appel, M.; Chacun, H.; Desmaële, D.; Taran, F.; Georgin, D.; Couvreur, P. Low-density lipoprotein receptor-mediated endocytosis of PEGylated nanoparticles in rat brain endothelial cells. Cell. Mol. Life Sci. 2007, 64, 356–364. [Google Scholar] [CrossRef] [PubMed]
- Basu, S.K.; Goldstein, J.L.; Anderson, R.G.; Brown, M.S. Monensin interrupts the recycling of low density lipoprotein receptors in human fibroblasts. Cell 1981, 24, 493–502. [Google Scholar] [CrossRef]
- Elbakry, A.; Zaky, A.; Liebl, R.; Rachel, R.; Goepferich, A.; Breunig, M. Layer-by-layer assembled gold nanoparticles for siRNA delivery. Nano Lett. 2009, 9, 2059–2064. [Google Scholar] [CrossRef] [PubMed]
- Miladi, I.; Alric, C.; Dufort, S.; Mowat, P.; Dutour, A.; Mandon, C.; Laurent, G.; Bräuer-Krisch, E.; Herath, N.; Jean-Luc Coll, J.-L.; et al. The in vivo radiosensitizing effect of gold nanoparticles based MRI contrast agents. Small 2014, 10, 1116–1124. [Google Scholar] [CrossRef] [PubMed]
- Xing, R.; Liu, K.; Jiao, T.; Zhang, N.; Ma, K.; Zhang, R.; Zou, Q.; Ma, G.; Yan, X. An Injectable Self-Assembling Collagen-Gold Hybrid Hydrogel for Combinatorial Antitumor Photothermal/Photodynamic Therapy. Adv. Mater. 2016, 28, 3669–3676. [Google Scholar] [CrossRef] [PubMed]
- Su, G.; Zhou, H.; Mu, Q.; Zhang, Y.; Li, L.; Jiao, P.; Jiang, G.; Yan, B. Effective surface charge density determines the electrostatic attraction between nanoparticles and cells. J. Phys. Chem. C 2012, 116, 4993–4998. [Google Scholar] [CrossRef]
- Elci, S.G.; Jiang, Y.; Yan, B.; Kim, S.T.; Saha, K.; Moyano, D.F.; Tonga, G.Y.; Jackson, L.C.; Rotello, V.M.; Vachet, R.W. Surface charge controls the suborgan biodistributions of gold nanoparticles. ACS Nano 2016, 10, 5536–5542. [Google Scholar] [CrossRef] [PubMed]
- Oh, N.; Park, J.-H. Surface chemistry of gold nanoparticles mediates their exocytosis in macrophages. ACS Nano 2014, 8, 6232–6241. [Google Scholar] [CrossRef] [PubMed]
- Tlotleng, N.; Vetten, M.A.; Keter, F.K.; Skepu, A.; Tshikhudo, R.; Gulumian, M. Cytotoxicity, intracellular localization and exocytosis of citrate capped and PEG functionalized gold nanoparticles in human hepatocyte and kidney cells. Cell Biol. Toxicol. 2016, 32, 305–321. [Google Scholar] [CrossRef] [PubMed]
- Nel, A.E.; Mädler, L.; Velegol, D.; Xia, T.; Hoek, E.M.; Somasundaran, P.; Klaessig, F.; Castranova, V.; Thompson, M. Understanding biophysicochemical interactions at the nano–bio interface. Nat. Mater. 2009, 8, 543–557. [Google Scholar] [CrossRef] [PubMed]
- Davis, C.G.; Goldstein, J.L.; Südhof, T.C.; Anderson, R.G.; Russell, D.W.; Brown, M.S. Acid-dependent ligand dissociation and recycling of LDL receptor mediated by growth factor homology region. Nature 1987, 326, 760. [Google Scholar] [CrossRef] [PubMed]
- Yamashiro, D.J.; Fluss, S.R.; Maxfield, F.R. Acidification of endocytic vesicles by an ATP-dependent proton pump. J. Cell Biol. 1983, 97, 929–934. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bonfleur, M.L.; Vanzela, E.C.; Ribeiro, R.A.; de Gabriel Dorighello, G.; de França Carvalho, C.P.; Collares-Buzato, C.B.; Carneiro, E.M.; Boschero, A.C.; de Oliveira, H.C.F. Primary hypercholesterolaemia impairs glucose homeostasis and insulin secretion in low-density lipoprotein receptor knockout mice independently of high-fat diet and obesity. Biochim. Biophys. Acta 2010, 1801, 183–190. [Google Scholar] [CrossRef] [PubMed]
- Brown, M.; Goldstein, J. A receptor-mediated pathway for cholesterol homeostasis. Science 1986, 232, 34–47. [Google Scholar] [CrossRef] [PubMed]
- Wang, W.; Ding, X.; Xu, Q.; Wang, J.; Wang, L.; Lou, X. Zeta-potential data reliability of gold nanoparticle biomolecular conjugates and its application in sensitive quantification of surface absorbed protein. Colloids Surf. B Biointerfaces 2016, 148, 541–548. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.; Zeiri, O.; Neyman, A.; Stellacci, F.; Weinstock, I.A. Nucleation and island growth of alkanethiolate ligand domains on gold nanoparticles. ACS Nano 2012, 6, 629–640. [Google Scholar] [CrossRef] [PubMed]
Mixed Charged Thiols (11-mercaptoundecanoic acid (MUA)/1-octanethiol (OT)) | Zeta Potential (mV) (Mean ± SEM) | Hydrodynamic Size (nm) (Mean ± SEM) |
---|---|---|
0:1 (0% MUA–AuNPs) | −20.21 ± 0.48 | 42.12 ± 1.32 |
1:2 (33% MUA–AuNPs) | −19.92 ± 0.43 | 42.23 ± 2.12 |
1:1 (50% MUA–AuNPs) | −16.06 ± 0.38 | 43.14 ± 1.24 |
2:1 (67% MUA–AuNPs) | −14.54 ± 0.52 | 42.67 ± 1.52 |
1:0 (100% MUA–AuNPs) | −33.14 ± 0.42 | 42.45 ± 2.38 |
Citrate–AuNPs | −32.05 ± 1.77 | 41.5 ± 2.42 |
© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Bai, X.; Zhang, J.; Chang, Y.-N.; Gu, W.; Lei, R.; Qin, Y.; Xia, S.; Ma, S.; Liang, Y.; Chen, K.; et al. Nanoparticles with High-Surface Negative-Charge Density Disturb the Metabolism of Low-Density Lipoprotein in Cells. Int. J. Mol. Sci. 2018, 19, 2790. https://doi.org/10.3390/ijms19092790
Bai X, Zhang J, Chang Y-N, Gu W, Lei R, Qin Y, Xia S, Ma S, Liang Y, Chen K, et al. Nanoparticles with High-Surface Negative-Charge Density Disturb the Metabolism of Low-Density Lipoprotein in Cells. International Journal of Molecular Sciences. 2018; 19(9):2790. https://doi.org/10.3390/ijms19092790
Chicago/Turabian StyleBai, Xue, Jiaxin Zhang, Ya-Nan Chang, Weihong Gu, Runhong Lei, Yanxia Qin, Shibo Xia, Sihan Ma, Yuelan Liang, Kui Chen, and et al. 2018. "Nanoparticles with High-Surface Negative-Charge Density Disturb the Metabolism of Low-Density Lipoprotein in Cells" International Journal of Molecular Sciences 19, no. 9: 2790. https://doi.org/10.3390/ijms19092790