氯化镧和柠檬酸镧在胃肠道以磷酸镧微粒为转化物种且主要通过M细胞进行转运吸收

doi:10.5246/jcps.2018.08.056

摘要/Abstract

摘要：

In the present study, we investigated the transformed species and the absorptive mechanism of rare earth elements (REEs) in gastrointestinal (GI) tract, using LaCl₃and LaCitas representative compounds.Artificial gastric and intestinal fluids were used to simulate the environment of the digestive tract in vivo. The inductively coupled plasma mass spectrometry (ICP-MS) result showed that more than 99.9% of LaCl₃and LaCit formed precipitation in artificial intestinal fluid, with the average size distribution of 200 nm (2-h incubation) increasing to 600 nm (24-h incubation) determined by dynamic light scattering (DLS), indicating the aggregation of the particles. The Fourier transform infrared spectroscopy (FTIR) analysis demonstrated that the constituents of these particles were mainly in the form of lanthanum phosphates. To explore the transport mechanism of REEs in GI tract, the mice Peyer's patches (PPs) and intestinal epithelium were separated to evaluate the content of lanthanum by ICP-MS following oral administration with 2 or 100 mg/kg/day of LaCit for 7 d. The results showed that the amount of lanthanum phosphate particlesabsorbed by PPs was significantly greater than that of intestinal epithelium, indicating that lanthanum particles might be phagocytosed mainly by M cells located in the follicle-associated epithelium (FAE) overlying PPs. Furthermore, Caco-2 cell monoculture and Caco-2/Raji B cell coculture models were established to simulate intestinal epithelial cells and FAE, respectively. The result showed that the transport of lanthanum in Caco-2/Raji B coculture model was significantly higher than that in Caco-2 monoculture model (about 60 times higher), and the level of lanthanum in the basal compartment of Caco-2 monoculture model was very low, supporting that M cells were the main route for lanthanum phosphate particles to be transported and absorbed. Taken together, these data suggested that LaCl₃and LaCit in GI tract were absorbed mainly via M cells with lanthanum phosphates as transformed species. The obtained results would provide the theoretical basis for the rational application of REEs in agriculture and medicine.

关键词: Lanthanum chloride (LaCl3), Lanthanum citrate (LaCit), Lanthanum phosphates, Artificial intestinal fluid, Peyer’s patches, M cells, Caco-2 cell monoculture model, Caco-2/Raji B cells coculture model, Transport and absorption

Abstract:

本工作以氯化镧(LaCl₃)和柠檬酸镧(LaCit)为代表性化合物, 对稀土在消化道内的物种存在形式及吸收转运机制进行了研究和探讨。研究采用了人工胃液和肠液模拟体内消化道环境。电感耦合等离子体质谱(ICP-MS)检测结果表明, 大于99.9%的La发生了沉淀。散射光粒度(DLS)分析结果显示, 这些微粒的平均粒径由200 nm (孵育2 h)增大到600 nm左右(孵育24 h), 这表明微粒之间发生了聚集。傅立叶变换红外光谱(FTIR)研究表明, 稀土微粒的主要成分为稀土磷酸盐。为研究稀土在肠道中的吸收转运机制, 以LaCit (2或100 mg/kg/day)对小鼠灌胃给药7天, 取小肠的普通肠段和派氏结进行了ICP-MS分析。派氏结中的La含量明显高于普通肠段, 这提示稀土微粒可能主要由派氏结上淋巴滤泡上皮中的M细胞吞噬转运。此外, 建立了Caco-2细胞单培养模型和 Caco-2/Raji B 细胞共培养模型分别模拟小肠上皮和滤泡上皮。结果显示, Caco-2/Raji B 细胞共培养模型转运的La含量显著高于Caco-2细胞单培养模型(大约60倍左右), 且 Caco-2 细胞单层透过的镧的量极低, 这说明M细胞是吸收转运磷酸镧的主要途径。以上结果表明氯化镧和柠檬酸镧在胃肠道主要以磷酸镧微粒形式通过M细胞进行转运吸收。本研究将为稀土在农业和医学中的合理应用提供科学依据。

Key words: 氯化镧, 柠檬酸镧, 磷酸镧, 人工肠液, 派氏结, M细胞, Caco-2 细胞单培养模型, Caco-2/Raji B 细胞共培养模型, 吸收转运

中图分类号:

R916.3

Supporting:

黄慧霞, 刘会雪, 马孝杰, 管辉, 杨晓改. 氯化镧和柠檬酸镧在胃肠道以磷酸镧微粒为转化物种且主要通过M细胞进行转运吸收[J]. 中国药学(英文版), 2018, 27(8): 553-564.

Huixia Huang, Huixue Liu, Xiaojie Ma, Hui Guan, Xiaogai Yang. Lanthanum chloride or citrate is absorbed mainly via M cells in gastrointestinal tracts with lanthanum phosphates as the transformed species[J]. Journal of Chinese Pharmaceutical Sciences, 2018, 27(8): 553-564.

参考文献

[1] Du, X.; Graedel, T.E. Uncovering the Global Life Cycles of the Rare Earth Elements. Sci. Rep. 2011, 1, 145.

[2] Gwenzi, W.; Mangori, L.; Danha, C.; Chaukura, N.; Dunjana, N.; Sanganyado, E. Sources, behaviour, and environmental and human health risks of high-technology rare earth elements as emerging contaminants. Sci. Total. Environ. 2018, 636, 299–313.

[3] Cheng, J.; Fei, M.; Fei, M.; Sang, X.; Sang, X.; Cheng, Z.; Gui, S.; Zhao, X.; Sheng, L.; Sun, Q.; Hu, R.; Wang, L.; Hong, F. Gene expression profile in chronic mouse liver injury caused by long-term exposure to CeCl3. Environ. Toxicol. 2014, 29, 837–846.

[4] Huang, P.; Li, J.; Zhang, S.; Chen, C.; Han, Y.; Liu, N.; Xiao, Y.; Wang, H.; Zhang, M.; Yu, Q.; Liu, Y.; Wang, W. Effects of lanthanum, cerium, and neodymium on the nuclei and mitochondria of hepatocytes: accumulation and oxidative damage. Environ. Toxicol. Pharmacol. 2011, 31, 25–32.

[5] Kawagoe, M.; Hirasawa, F.; Cun Wang, S.; Liu, Y.; Ueno, Y.; Sugiyama, T. Orally administrated rare earth element cerium induces metallothionein synthesis and increases glutathione in the mouse liver. Life. Sci. 2005, 77, 922–937.

[6] Pagano, G.; Aliberti, F.; Guida, M.; Oral, R.; Siciliano, A.; Trifuoggi, M.; Tommasi, F. Rare earth elements in human and animal health: State of art and research priorities. Environ. Res. 2015, 142, 215–220.

[7] Li, N.; Cheng, J.; Cheng, Z.; Hu, R.; Cai, J.; Gao, G.; Cui, Y.; Wang, L.; Hong, F. Molecular mechanism of inflammatory response in mouse liver caused by exposure to CeCl3. Environ. Toxicol. 2013, 28, 349–358.

[8] Lacour, B.; Lucas, A.; Auchère, D.; Ruellan, N.; Patey, N.M.D.S.; Drüeke, T.B. Chronic renal failure is associated with increased tissue deposition of lanthanum after 28-day oral administration. Kidney Int. 2005, 67, 1062–1069.

[9] Pennick, M.; Dennis, K.; Damment, S.J. Absolute bioavailability and disposition of lanthanum in healthy human subjects administered lanthanum carbonate. J. Clin. Pharmacol. 2006, 46, 738–746.

[10] Feng, M.; Fan, Y.Z.; Ma, X.J.; Li, J.X.; Yang, X.G. The gadolinium-based contrast agent Omniscan® promotes in vitro fibroblast survival through in situ precipitation. Metallomics. 2015, 7, 1103–1120.

[11] Idée, J.M.; Port, M.; Raynal, I.; Schaefer, M.; Greneur, S.L.; Corot, C. Clinical and biological consequences of transmetallation induced by contrast agents for magnetic resonance imaging: a review. Fundam. Clin. Pharmacol. 2006, 20, 563–576.

[12] Abraham, J.L.; Thakral, C.; Skov, L.; Rossen, K.; Marckmann, P. Dermal inorganic gadolinium concentrations: evidence for in vivo transmetallation and long-term persistence in nephrogenic systemic fibrosis. Br. J. Dermatol. 2008, 158, 273–280.

[13] Florence, A.T. The oral absorption of micro- and nanoparticulates: neither exceptional nor unusual. Pharm. Res. 1997, 14, 259–266.

[14] Etienne-Mesmin, L.; Chassaing, B.; Sauvanet, P.; Denizot, J.; Blanquet-Diot, S.; Darfeuille-Michaud, A.; Pradel, N.; Livrelli, V. Interactions with M cells and macrophages as key steps in the pathogenesis of enterohemorrhagic Escherichia coli infections. PLoS. One. 2011, 6, e23594.

[15] Owen, R.L.; Pierce, N.F.; Apple, R.T.; Cray, W.C. Jr. M cell transport of Vibrio cholerae from the intestinal lumen into Peyer's patches: a mechanism for antigen sampling and for microbial transepithelial migration. J. Infect. Dis. 1986, 153, 1108–1118.

[16] Ermak, T.H.; Dougherty, E.P.; Bhagat, H.R.; Kabok, Z.; Pappo, J. Uptake and transport of copolymer biodegradable microspheres by rabbit Peyer’s patch M cells. Cell. Tissue. Res. 1995, 279, 433–436.

[17] Des Rieux, A.; Fievez, V.; Theate, I.; Mast, J.; Preat, V.; Schneider, Y.J. An improved in vitro model of human intestinal follicle-associated epithelium to study nanoparticle transport by M cells. Eur. J. Pharm. Sci. 2007, 30, 380–391.

[18] Pappo, J.; Ermak, T.H. Uptake and translocation of fluorescent latex particles by rabbit Peyer’s patch follicle epithelium: a quantitative model for M cell uptake. Clin. Exp. Immunol. 1989, 76, 144–148.

[19] Des Rieux, A.; Ragnarsson, E.G.; Gullberg, E.; Preat, V.; Schneider, Y.J.; Artursson, P. Transport of nanoparticles across an in vitro model of the human intestinal follicle associated epithelium. Eur. J. Pharm. Sci. 2005, 25, 455–465.

[20] Brun, E.; Barreau, F.; Veronesi, G.; Fayard, B.; Sorieul, S.; Chaneac, C.; Carapito, C.; Rabilloud, T.; Mabondzo, A.; Herlin-Boime, N.; Carriere, M. Titanium dioxide nanoparticle impact and translocation through ex vivo, in vivo and in vitro gut epithelia. Part. Fibre. Toxicol. 2014, 11, 13.

[21] Liu, T.Y.; Chen, S.Y.; Liu, D.M.; Liou, S.C. On the study of BSA-loaded calcium-deficient hydroxyapatite nano-carriers for controlled drug delivery. J. Control. Release. 2005, 107, 112–121.

[22] Shakweh, M.; Ponchel, G.; Fattal, E. Particle uptake by Peyer’s patches: a pathway for drug and vaccine delivery. Expert. Opin. Drug Deliv. 2004, 1, 141–163.

[23] Jepson, M.A.; Simmons, N.L.; Savidge, T.C.; James, P.S.; Hirst, B.H. Selective binding and transcytosis of latex microspheres by rabbit intestinal M cells. Cell Tissue. Res. 1993, 271, 399–405.

[24] Lefevre, M.E.; Hancock, D.C.; Joel, D.D. Intestinal barrier to large particulates in mice. J. Toxicol. Environ. Health. 1980, 6, 691–704.

[25] Jepson, M.; Simmons, N.; O''Hagan, D.; Hirst, B. Comparison of Poly(DL-Lactide-co-Glycolide) and Polystyrene Microsphere Targeting to Intestinal M Cells. J. Drug Target. 1993, 1, 245–249.

[26] Jani, P.; Halbert, G.W.; Langridge, J.; Florence, A.T. The uptake and translocation of latex nanospheres and microspheres after oral administration to rats. J. Pharm. Pharmacol. 1989, 41, 809–812.

[27] Smith, M.W.; Peacock, M.A. “M” cell distribution in follicle-associated epithelium of mouse Peyer’s patch. Am. J. Anat. 1980, 159, 167–175.

[28] Pappo, J.; Steger, H.J.; Owen, R.L. Differential adherence of epithelium overlying gut-associated lymphoid tissue. An ultrastructural study. Lab. Invest. 1988, 58, 692–697.

[29] Gamboa, J.M.; Leong, K.W. In vitro and in vivo models for the study of oral delivery of nanoparticles. Adv. Drug Deliv. Rev. 2013, 65, 800–810.

[30] Yang, X.G.; Fan, Y.Z.; Feng, M.; Liu, H.X.; Wang, K. Chemical species and mechanisms underlying biological effects of rare earth elements. Sci. Sin. 2014, 44, 521–530.

[31] Waring, P.M.; Watling, R.J. Rare earth deposits in a deceased movie projectionist. A new case of rare earth pneumoconiosis? Med. J. Aust. 1990, 153, 726–730.

[32] High, W.A.; Ayers, R.A.; Cowper, S.E. Gadolinium is quantifiable within the tissue of patients with nephrogenic systemic fibrosis. J. Am. Acad. Dermatol. 2007, 56, 710–712.

[33] Jepson, M.A.; Simmons, N.L.; Savidge, T.C.; James, P.S.; Hirst, B.H. Selective binding and transcytosis of latex microspheres by rabbit intestinal M cells. Cell. Tissue. Res. 1993, 271, 399–405.

[34] Kalgaonkar, S.; Lonnerdal, B. Receptor-mediated uptake of ferritin-bound iron by human intestinal Caco-2 cells. J. Nutr. Biochem. 2009, 20, 304–311.

[35] Jani, P.; Halbert, G.W.; Langridge, J.; Florence, A.T. Nanoparticle Uptake by the Rat Gastrointestinal Mucosa: Quantitation and Particle Size Dependency. J. Pharm. Pharmacol. 1990, 42, 821–826.

[36] Des, R.A.; Ragnarsson, E.G.; Gullberg, E.; Préat, V.; Schneider, Y. J.; Artursson, P. Transport of nanoparticles across an in vitro model of the human intestinal follicle associated epithelium. Eur. J. Pharm. Sci. 2005, 25, 455–465.

[37] Corr, S.C.; Gahan, C.C.G.M.; Hill, C., Corr, S.C., Gahan, C.C.; Hill, C. M-cells: origin, morphology and role in mucosal immunity and microbial pathogenesis. FEMS. Immunol. Med. Microbiol. 2008, 52, 2–12.

[38] Wells, C.L.; Maddaus, M.A.; Erlandsen, S.L.; Simmons, R.L. Evidence for the phagocytic transport of intestinal particles in dogs and rats. Infect. Immun. 1988, 56, 278–282.

[39] Lomotan, E.A.; Brown, K.A.; Speaker, T.J. Offita, P.A., Aqueous-based microcapsules are detected primarily in gut-associated dendritic cells after oral inoculation of mice. Vaccine. 1997, 15, 1959–1962.

[40] Eberl, G.; Lochner, M. The development of intestinal lymphoid tissues at the interface of self and microbiota. Mucosal. Immunol. 2009, 2, 478–485.

[41] Kimura, S. Molecular insights into the mechanisms of M-cell differentiation and transcytosis in the mucosa-associated lymphoid tissues. Anat. Sci. Int. 2018, 93, 23–34.

[42] Khan, I.U.; Huang, J.; Liu, R.; Wang, J.; Xie, J.; Zhu, N. Phage Display-Derived Ligand for Mucosal Transcytotic Receptor GP-2 Promotes Antigen Delivery to M Cells and Induces Antigen-Specific Immune Response. SLAS. Discov. 2017, 22, 879–886.