镰形棘豆黄酮含药血清对人肺癌A549细胞活力、凋亡及周期的影响

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英文篇名：Effect of Medicated Serum Containing Flavonoids from Oxytropis falcata on Viability, Apoptosis and Life Cycle of Human Lung Adenocarcinoma A549 Cells In Vitro 作者：牛俐燃 ; 李冠聪 ; 张婉婷 ; 杨光明 ; 潘扬 英文作者：NIU Liran;LI Guancong;ZHANG Wanting;YANG Guangming;PAN Yang;School of Pharmacy,Nanjing University of Chinese Medicine;Key Laboratory of State Administration of Traditional Chinese Medicine for Standardization of Chinese Medicine Processing & Jiangsu Key Laboratory of Chinese Medicine Processing;Laboratory of Medical Fungi and Phyto-Biotech,Nanjing University of Chinese Medicine; 关键词：镰形棘豆 ; 黄酮提取物 ; 含药血清 ; A549细胞 ; 细胞活力 ; 细胞凋亡 ; 细胞周期 英文关键词：Oxytropis falcata;;flavonoids;;medicated serum;;A549 cells;;cell viability;;cell apoptosis;;cell cycle 中文刊名：ZYXY 英文刊名：Traditional Chinese Drug Research and Clinical Pharmacology 机构：南京中医药大学药学院;江苏省中药炮制重点实验室国家中医药管理局中药炮制标准重点研究室;南京中医药大学药用菌与中药生物技术研究所; 出版日期：2019-07-16 出版单位：中药新药与临床药理 年：2019 期：v.30;No.160 基金：国家自然科学基金项目(81473420,30902012);; 江苏高校优势学科建设工程资助项目(PAPD);; 江苏省高校品牌专业建设工程资助项目(PPZY2015A070) 语种：中文; 页：ZYXY201907007 页数：6 CN：07 ISSN：44-1308/R 分类号：36-41

摘要

目的研究镰形棘豆黄酮提取物(OFF)含药血清对人肺腺癌A549细胞活力、细胞形态学、凋亡率及增殖周期的影响。方法将SD大鼠随机分为5组:空白对照组,阳性对照组(环磷酰胺CTX,40 mg·kg~(-1)),OFF低剂量组(0.587 g·kg~(-1))、中剂量组(1.174 g·kg~(-1))、高剂量组(2.348 g·kg~(-1));早晚各给药1次,连续4 d,制备含药血清。采用CCK-8法检测细胞活力;倒置显微镜观察细胞形态;Annexin V-FITC/PI双染法检测细胞凋亡率;流式细胞术检测细胞周期。结果在12、24、48 h给药时间内,OFF含药血清各剂量组的细胞活力随剂量递增呈抑制增强效果,且在24 h抑制作用最强,48 h抑制作用减弱,但与空白对照组比较,差异均有统计学意义(P <0.001);倒置显微镜下观察, OFF含药血清作用24 h后,各剂量组的A549细胞均生长缓慢,细胞呈不规则形态。与空白对照组比较,OFF含药血清低、中、高剂量组的凋亡细胞比例上升,凋亡率分别为47.91%、55.09%、60.37%,差异均有统计学意义(P <0.001);OFF各剂量组的细胞周期中,G0/G1期细胞比例明显上升(P <0.01),S期(P<0.05,OFF高剂量组)及G2期(P<0.01,OFF中、高剂量组)比例下降。结论OFF含药血清对A549人肺腺癌细胞活力有较好的抑制作用,并能够诱导细胞凋亡,将细胞周期阻滞在G0/G1期。
        Objective To study the effects of serum containing flavonoids from Oxytropis falcata(OFF) on cell viability,cell morphology,apoptosis rate,proliferation cycles of human lung adenocarcinoma A549 cells by serum pharmacology. Methods SD rats were randomly divided into blank control, positive control(cyclophosphamide CTX,40 mg·kg~(-1)),low-dose(0.587 g·kg~(-1)),medium-dose(1.174 g·kg~(-1))and high-dose(2.348 g·kg~(-1))of OFF groups. Each group was administered intragastrically every 12 h for 4 days. OFF-medicated serum samples were then extracted for A549 cell experiments. The cell viability was investigated by CCK-8 assay, cell morphology was observed by inverted microscope,apoptosis was analyzed by Annexin V-FITC/propidium iodide(PI)staining,and flow cytometry was adopted to detect proliferation cycles of cells. Results After being treated with OFF-medicated serum for 12, 24 and 48 h, cell viabilities of A549 cells were inhibited in a dose dependent manner; and the inhibition reached the top at 24 h while weakened in 48 h. A549 cells showed slower growth and irregular morphology after OFF serum treatment for 24 h. Compared with the control group,the proportion of apoptotic cells increased significantly in all treatment groups, with the apoptosis rates of 47.91%, 55.09% and 60.37%,respectively(P < 0.001). Meanwhile,the proportion of cells at G0/G1 phase also increased(P < 0.01),while the proportion of cells at S phase(P < 0.05,OFF high dose group)and G2 phase decreased(P < 0.01,OFF high and medium dose groups). Conclusion The serum containing OFF can inhibit the viability of A549 human lung adenocarcinoma cells in vitro,induce apoptosis and arrest cell proliferation at G0/G1 phase.

引文

[1]中华人民共和国卫生部.中华人民共和国卫生部药品标准:藏药(第一册)[S].1995:340.
    [2]帝玛尔·丹增彭措.晶珠本草[M].上海:上海科学技术出版社,1986.
    [3]CHEN W H,WU Q X,WANG R,et al.Oxytrofalcatins A-F,N-benzoylindole analogues from the roots of Oxytropis falcata(Leguminosae)[J].Phytochemistry,2010,71(8-9):1002-1006.
    [4]姚淑英,马云保,唐亚,等.镰形棘豆的化学成分研究[J].中国中药杂志,2008,33(12):1418-1421.
    [5]顾青,蔡银娜,杨光明,等.藏药镰形棘豆的化学成分及其抗肿瘤活性研究[J].中国实验方剂学杂志,2013,19(11):72-75.
    [6]陈锦珊,杨钦磊,刘晓玲,等.藏药镰形棘豆的化学成分、药理作用及毒理学研究进展[J].中国药房,2016,27(28):3945-3948.
    [7]高博,李钦,杨丽霞,等.镰形棘豆的研究现状与分析[J].中医研究,2017,30(1):75-78.
    [8]程海清,牛俐燃,李冠聪,等.镰形棘豆脂溶性生物碱含药血清对人肺癌A549细胞活力、凋亡及周期的影响[J].中药新药与临床药理,2018,29(5):535-539.
    [9]陈醒,杨光明,张芳芳,等.镰形棘豆诱导人肝癌细胞SMMC-7721凋亡及初步的机制研究[J].中国中药杂志,2011,36(10):1362-1365.
    [10]WANG D,TANG W,YANGG M,et al.Anti-inflammatory,antioxidant and cytotoxic activities of flavonoids from Oxytropis falcata Bunge[J].Chinese Journal of Natural Medicines,2010,8(6):461-465.
    [11]楼成华,王明艳,杨欢,等.镰形棘豆中黄酮类化合物抗肿瘤活性的体外实验研究[J].南京中医药大学学报,2009,25(1):46-50.
    [12]赵万红,曹永孝,袁泽飞.中药血清药理学的方法学探讨[J].中药新药与临床药理,2002,13(2):122-124.
    [13]李萌.镰形棘豆抗肿瘤作用体内体外活性部位筛选及基于TLRs/NF-κB信号通路的机制研究[D].南京:南京中医药大学,2017.
    [14]李萌,程海清,杨光明,等.藏药镰形棘豆醇提物酸碱分离及抗小鼠Lewis肺癌活性初步筛选[J].中药新药与临床药理,2017,28(5):628-632.
    [15]TORRE L A,BRAY F,SIEGEL R L,et al.Global cancer statistics,2012[J].A Cancer Journal for Clinicians,2015,65(2):87-108.
    [16]YUAN I,HORNG C T,CHEN V C,et al.Escitalopram oxalate inhibits proliferation and migration and induces apoptosis in nonsmall cell lung cancer cells[J].Oncology Letters,2018,15(3):3376-3382.
    [17]CHIKARA S,LINDSEY K,DHILLON H,et al.Enterolactone Induces G1-phase cell cycle arrest in nonsmall cell lung cancer cells by downregulating cyclins and cyclin-dependent kinases[J].Nutrition and Cancer,2017,69(4):652-662.
    [18]杨光明,燕珂,顾青,等.藏药镰形棘豆总黄酮对SMMC-7721肝癌细胞凋亡及Bcl-2,Bax蛋白表达的影响[J].中国药学杂志,2013,48(24):2113-2116.
    [19]张芳芳,杨光明,王栋,等.镰形棘豆提取物抗肿瘤活性的体外实验研究[J].药学与临床研究,2010,18(2):135-138.
    [20]吴沅皞,刘维,赵文甲.血清药理学方法对药理、药效学和新药研发的贡献[J].中国组织工程研究,2018,22(24):3914-3920.
    [21]李黎,王明艳,陈海彬,等.消癌解毒方含药血清对人肝癌SMMC-7721细胞的抑制作用[J].中药新药与临床药理,2011,22(6):587-589.