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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

Effect of Medicated Serum Containing Flavonoids from Oxytropis falcata on Viability, Apoptosis and Life Cycle of Human Lung Adenocarcinoma A549 Cells In Vitro

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【作者】 牛俐燃李冠聪张婉婷杨光明潘扬

【Author】 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;

【通讯作者】 杨光明;潘扬;

【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

【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。

【Abstract】 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.

【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)
  • 【文献出处】 中药新药与临床药理 ,Traditional Chinese Drug Research and Clinical Pharmacology , 编辑部邮箱 ,2019年07期
  • 【分类号】R285
  • 【被引频次】2
  • 【下载频次】351
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