目的 制备雄黄纳米粒并考察其理化性质,研究雄黄纳米粒对人肺癌细胞A549增殖凋亡的影响。方法 以聚乙烯吡咯烷酮K30(PVP)为载体,通过生物模板法制备雄黄纳米粒。采用透射电子显微镜(transmission electron microscope,TEM)、红外光谱分析、紫外光谱分析以及粒度分析等方法对所制备的纳米粒进行表征;原子吸收光谱法测定纳米粒中As4S4的载药量和包封率;噻唑蓝(MTT)法考察雄黄纳米粒对A549细胞的体外抑制活性,并计算半数抑制浓度(IC50),Annexin V/PI双染色法检测A549细胞的凋亡,Western blot法检测survivin蛋白的表达。结果 所制备的雄黄纳米粒为形态规则、分散均匀的球形粒子,纳米粒的粒径为50.7 nm左右,电位为-23.3 mV,冻干复溶后纳米粒粒径为68.1 nm左右,电位为-21.4 mV,As4S4@PVP NPs的包封率为25.9%,载药量为13.4%,紫外光谱显示As4S4@PVP NPs中出现PVP的特征吸收峰,红外光谱分析显示As4S4@PVP NPs与PVP的红外光谱峰位一致,As4S4@PVP NPs在H2O、PBS、RPMI-1640完全培养基条件下的粒径48 h内无明显变化,体外结果显示,纳米雄黄可显著抑制A549细胞的增殖,与对照组相比较,As4S4@PVP NPs能有效诱导细胞凋亡,降低survivin蛋白的表达。结论 成功制备雄黄纳米粒,所制纳米粒粒径小且稳定,可通过降低survivin的表达,有效诱导肺癌A549细胞发生凋亡。
Abstract
OBJECTIVE To prepare realgar nanoparticles and investigate their physicochemical properties, study the effect of realgar nanoparticles on the proliferation and apoptosis of human lung cancer cell A549. METHODS Using polyvinylpyrrolidone K30 (PVP) as a carrier, realgar nanoparticles were prepared by the biological template method. The prepared nanoparticles were characterized by transmission electron microscope (transmission electron microscope,TEM), infrared spectroscopy, ultraviolet spectroscopy and particle size analysis, and the drug loading and entrapment efficiency of As4S4 in the nanoparticles were determined by atomic absorption spectrometry. The inhibitory activity of realgar nanoparticles on A549 cells in vitro was detected by thiazolyl (MTT) method, and the half inhibitory concentration (IC50) was calculated. The apoptosis of A549 cells was detected by Annexin V/PI double staining, and the expression of survivin protein was detected by Western blot. RESULTS The prepared realgar nanoparticles are spherical particles with regular morphology and uniform dispersion. The size of the nanoparticles is about 50.7 nm and the potential is -23.3 mV. After freeze-drying and reconstitution, the size of the nanoparticles is about 68.1 nm and the potential is -21.4 mV, the encapsulation rate of As4S4@PVP NPs is 25.9%, and the drug loading is 13.4%. The ultraviolet spectrum shows that the characteristic absorption peak of PVP appears in As4S4@PVP NPs, and the infrared spectrum analysis shows the infrared spectrum peaks of As4S4@PVP NPs and PVP. The position of As4S4@PVP NPs did not change significantly within 48 h under the conditions of H2O, PBS, and RPMI-1640 complete media. The in vitro results showed that nano realgar can significantly inhibit the proliferation of A549 cells. Compared with the control group, As4S4 @PVP NPs can effectively induce cell apoptosis and reduce the expression of survivin protein. CONCLUSION Realgar nanoparticles are successfully prepared, and the size of the nanoparticles is small and stable, which could effectively induce apoptosis of lung cancer A549 cells by reducing the expression of survivin.
关键词
纳米雄黄 /
聚乙烯吡咯烷酮 /
细胞凋亡 /
survivin
{{custom_keyword}} /
Key words
nanometer realgar /
PVP /
cell apoptosis /
survivin
{{custom_keyword}} /
中图分类号：
R944
R965
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] ALLEMANI C, MATSUDA T, DI CARLO V, et al. Global surveillance of trends in cancer survival 2000-14 (CONCORD-3): analysis of individual records for 37 513 025 patients diagnosed with one of 18 cancers from 322 population-based registries in 71 countries[J]. Lancet, 2018, 391(10125):1023-1075.
[2] WANG J, YU F, SHANG Y, et al. Thyroid cancer: incidence and mortality trends in China, 2005-2015[J]. Endocrine, 2020, 68(1):163-173.
[3] GUO T, HUANG J, WAN X, et al. lncRNA metastasis-associated lung adenocarcinoma transcript 1 promotes proliferation and invasion of non-small cell lung cancer cells via down-regulating miR-202 expression[J]. Cell, 2020, 22(3):375-385.
[4] XU Y, ZHAO X H. IAP family and IAP-targeted cancer therapy[J]. Chin Bull Life Sci(生命科学), 2010, 22(2):161-168.
[5] HAJAZIMIAN S, MALEKI M, MEHRABAD S D, et al. Human Wharton′s jelly stem cells inhibit endometriosis through apoptosis induction[J]. Reproduction, 2020, 159(5):549-558.
[6] SONG Y. Comparative study of expression of Survivin protein in SCLC and PESC[D]. Zhengzhou: Zhengzhou University, 2007.
[7] WANG J, DING L, ZHOU J, et al. Target lipidomics approach to reveal the resolution of inflammation induced by Chinese medicine combination in Liu-Shen-Wan against realgar overexposure to rats[J]. J Ethnopharmacol, 2019, 249:112171.
[8] VERMEULEN M, PALKA K, VLCEK M, et al. Study of dry-and wet-process amorphous arsenic sulfides: synthesis, Raman reference spectra, and identification in historical art materials[J]. J Raman Spect, 2019, 50(3):396-406.
[9] ZHAO Y, YUAN B, ONDA K, et al. Anticancer efficacies of arsenic disulfide through apoptosis induction, cell cycle arrest, and pro-survival signal inhibition in human breast cancer cells[J]. Am J Cancer Res, 2018, 8(3):366-386.
[10] YANG Y. The apoptosis of lung cancer and liver cancer cells and their Objective: cancer stem cells induced by realgar nanoparticle[D]. Lanzhou: Lanzhou University, 2010.
[11] TIAN Y, WANG X, XI R, et al. Enhanced antitumor activity of realgar mediated by milling it to nanosize[J]. Int J Nanomed, 2014, 9:745-757.
[12] EMADI A, GORE S D.Arsenic trioxide-an old drug rediscovered[J]. Blood Rev, 2010, 24(4-5):191-199.
[13] WU J, SHAO Y, LIU J, et al. The medicinal use of realgar (As4S4) and its recent development as an anticancer agent[J]. J Ethnopharmacol, 2011, 135(3):595-602.
[14] CHENG P P. The Preparation of Realgar NanoParticle and the Studies of Inducing Apoptosis in MCF-7 Cells[D]. Shijiazhuang: Hubei University of Chinese Medicine, 2016.
[15] WANG X B, XI R G, LI Z L, et al. Preparation of nanoparticle realgar powders[J]. Pharm J Chin PLA(解放军药学学报), 2002, 18(3):129-134.
[16] HADDADI P, MANDAVI M, RAHBARGHAZI R. Down-regulation of Bcl2 and survivin, and up-regulation of Bax involved in copper (Ⅱ) phenylthiosemicarbazone complex-induced apoptosis in leukemia stem-like KG1a cells[J]. Proc Biochem, 2020, 92:190-196.
[17] KUAI Z Y, LI X X, LI Q W. Synthesis and biological evaluation of oleanolic acid analogs based on survivin target[J]. Chin Pharm J(中国药学杂志), 2018, 53(17):1438-1446.
[18] GU C, XU S L, TANG W W, et al. HeLa cell apoptosis induced by quercetin and effects of caspase-3, caspase-8 activation on apoptosis[J]. Chin Pharm J(中国药学杂志), 2011, 46(8):595-599.
[19] DENG W, SUN X W. The research progress of survivin targeted therapy in cancer research[J]. Pract Oncol J(实用肿瘤学杂志), 2014, 28(2):165-169.
[20] SUN Y, LI Y. Relationship between survivin and biological behavior of cancer[J]. J Int Oncol(国际肿瘤学杂志), 2011, 38(1):6-8.
[21] NING H M, WANG J, SU Y F, et al. Newly diagnosed acute myeloid leukemia patients-derived bone marrow mesenchymal stem cells suppress daunorubicin induced HL-60 cell apoptosis via modulating caspase-3/survivin[J]. J Exper Hematol(中国实验血液杂志), 2019, 27(6):1736-1743.
[22] XU J W, CHENG B. Advances in molecular mechanisms of elemene induced apoptosis in tumor cells[J]. J China Pharm(中国药房), 2013, 24(35):3348-3350.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
湖南省高层次卫生人才“225”工程项目资助(湘卫函〔2019〕196号);湖南中医药大学中医学国内一流建设学科项目资助(湘教通[2018]469号);湖南省中医药科研计划项目资助(2016130)
{{custom_fund}}
PDF(1908 KB)
