表没食子儿茶素没食子酸酯纳米颗粒的制备及其体外抗白血病效应研究
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摘要
目的:制备表没食子儿茶素没食子酸酯(EGCG)-壳聚糖(CS)纳米颗粒,并比较EGCG纳米颗粒与原料药对人急性淋巴细胞白血病细胞系Jurkat细胞的抑制作用。方法:选取壳聚糖(CS)为包封材料,采用注入-超声法将EGCG装载到CS形成的纳米粒中,制备出EGCG纳米颗粒。使用激光粒度仪对纳米EGCG的粒径和Zeta电位进行测定,采用场发射扫描电子显微镜观察其形态结构。采用CCK-8实验检测EGCG纳米颗粒抑制Jurkat细胞增殖的作用,采用流式细胞术检测其对Jurkat细胞凋亡的影响,比较EGCG纳米颗粒与原料药的体外抗白血病效应。结果:最优条件下制备的EGCG纳米颗粒包封率为(76±4)%,平均粒径为(116±25)nm,平均Zeta电位为(61.2±1.8)mV,具有球形微观结构。EGCG纳米颗粒抑制Jurkat细胞增殖作用及促Jurkat细胞凋亡作用显著高于EGCG原料药。结论:EGCG纳米颗粒在体外抗白血病效应明显优于原料药,为进一步探索其体内抗白血病效应提供了依据。
OBJECTIVE To prepare the chitosan encapsulated epigallocatechin-3-gallate(EGCG) nanoparticles(CS-NPs) and compare the inhibitory effect on human T-acute lymphoblastic leukemia cell line Jurkat of EGCG-CS-NPs and EGCG drug substance. METHODS Chitosan(CS) was selected as the encapsulation material, and EGCG was loaded into the nanoparticles formed by CS by injection-ultrasonic method to prepare EGCG nanoparticles.The particle size and zeta potential of nano-EGCG were measured by a laser particle size analyzer, and the morphological structure was observed using a field emission scanning electron microscope. In vitro anti-leukemia effects of EGCG nanoparticles and APIs were compared, emploging CCK-8 assay to examine the inhibitory effect of EGCG nanoparticles on the proliferation of Jurkat cells, and flow cytometry to detect its effect on the apoptosis of Jurkat cells. RESULTS The encapsulation rate of EGCG nanoparticles prepared under optimal conditions was(76±4)%, the average particle size was(116±25) nm, and the average zeta potential was(61.2±1.8) mV, with a spherical microstructure. EGCG-CS-NPs exhibited a significant higher anti-proliferation and pro-apoptotic effect on Jurkat cells compared to EGCG drug substance. CONCLUSION EGCG-CS-NPs exhibit a superior anti-leukemia effect in vitro, which lays a solid foundation for future research in vivo.
OBJECTIVE To prepare the chitosan encapsulated epigallocatechin-3-gallate(EGCG) nanoparticles(CS-NPs) and compare the inhibitory effect on human T-acute lymphoblastic leukemia cell line Jurkat of EGCG-CS-NPs and EGCG drug substance. METHODS Chitosan(CS) was selected as the encapsulation material, and EGCG was loaded into the nanoparticles formed by CS by injection-ultrasonic method to prepare EGCG nanoparticles.The particle size and zeta potential of nano-EGCG were measured by a laser particle size analyzer, and the morphological structure was observed using a field emission scanning electron microscope. In vitro anti-leukemia effects of EGCG nanoparticles and APIs were compared, emploging CCK-8 assay to examine the inhibitory effect of EGCG nanoparticles on the proliferation of Jurkat cells, and flow cytometry to detect its effect on the apoptosis of Jurkat cells. RESULTS The encapsulation rate of EGCG nanoparticles prepared under optimal conditions was(76±4)%, the average particle size was(116±25) nm, and the average zeta potential was(61.2±1.8) mV, with a spherical microstructure. EGCG-CS-NPs exhibited a significant higher anti-proliferation and pro-apoptotic effect on Jurkat cells compared to EGCG drug substance. CONCLUSION EGCG-CS-NPs exhibit a superior anti-leukemia effect in vitro, which lays a solid foundation for future research in vivo.
引文
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[13] Siddiqui IA,Bharali DJ, Nihal M, et al. Excellent anti-proliferative and pro-apoptotic effects of (-)-epigallocatechin-3-gallate encapsulated in chitosan nanoparticles on human melanoma cell growth both in vitro and in vivo [J]. Nanomedicine, 2014, 10(8): 1619-1626.
[14] Zhu M,Chen Y,Li R. Oral absorption and bioavailability of tea catechins [J]. Planta Med, 2000, 66(5): 444-447.
[15] Zeng L,Yan J,Luo L, et al. Preparation and characterization of (-)-Epigallocatechin-3-gallate (EGCG)-loaded nanoparticles and their inhibitory effects on Human breast cancer MCF-7 cells [J]. Sci Rep, 2017, 7: 45521.
[2] Kraszewska MD, Dawidowska M, Szczepański T, et al. T-cell acute lymphoblastic leukemia: recent molecular biology findings [J]. Br J Haematol, 2012, 156(3): 303-315.
[3] Islam MA. Cardiovascular efects of green tea catechins: Progress and promise [J]. Recent Pat Cardiovasc Drug Discov,2012, 7(2):88-99.
[4] Du X, Huang H, Li Q. Advances in anti-tumor effect of catechins EGCG and its mechanism [J]. Sci Tech Food Ind(食品工业科技), 2017, 38(24):308-314.
[5] Han Y, Chu J, Ge Z. Protective effect of (-)-epigallocatechin gallate on cardiovascular health [J]. Chin J Pathophysiol(中国病理生理杂志), 2012, 28(8):1521-1525.
[6] Gong Y, Zhang W, Liu W, et al. Protective effects of EGCG in experimental autoimmune encephalomyelitis mice [J]. Immunol J(免疫学杂志), 2017, 33(5):376-382.
[7] He M, Liu M, Wang S, et al. Research on EGCG improving the degenerative changes of the brain in AD model mice induced with chemical drugs [J]. J Chin Med Mater(中草药), 2012, 35(10):1641-1644.
[8] Masome GP, Batool P, Hedayatollah S, et al. Effect of Epigallocatechin-3-gallate (EGCG) on cell proliferation inhibition and apoptosis induction in lymphoblastic leukemia cell line [J]. J Herb Med Pharmacol, 2015, 4(2): 65-68.
[9] Li J, Wang X. Binding of (-)-epigallocatechin-3-gallate with thermally-induced bovine serum albumin/iota-carrageenan particles [J]. Food Chem, 2015, 168(8): 566-571.
[10] Hu B, Wang Y, Xie M, et al. Polymer nanoparticles composed with gallic acid grafted chitosan and bioactive peptides combined antioxidant, anticancer activities and improved delivery property for labile polyphenols [J]. J Funct Foods, 2015, 15:593-603.
[11] Delie F,Blanco-Príeto M J. Polymeric particulates to improve oral bioavailability of peptide drugs [J]. Molecules, 2005, 10(1): 65-80.
[12] Dube A,Nicolazzo JA,Larson I. Chitosan nanoparticles enhance the intestinal absorption of the green tea catechins (+)-catechin and (-)-epigallocatechin gallate [J]. Eur J Pharm Sci, 2010, 41(2): 219-225.
[13] Siddiqui IA,Bharali DJ, Nihal M, et al. Excellent anti-proliferative and pro-apoptotic effects of (-)-epigallocatechin-3-gallate encapsulated in chitosan nanoparticles on human melanoma cell growth both in vitro and in vivo [J]. Nanomedicine, 2014, 10(8): 1619-1626.
[14] Zhu M,Chen Y,Li R. Oral absorption and bioavailability of tea catechins [J]. Planta Med, 2000, 66(5): 444-447.
[15] Zeng L,Yan J,Luo L, et al. Preparation and characterization of (-)-Epigallocatechin-3-gallate (EGCG)-loaded nanoparticles and their inhibitory effects on Human breast cancer MCF-7 cells [J]. Sci Rep, 2017, 7: 45521.