目的 观察鸦胆子油对肺癌 A549 细胞凋亡和血管内皮生长因子分泌的影响，以探讨其治疗肺癌的作用机制。 方法 用不同浓度鸦胆子油乳（ 0.5 ， 1.25 ， 2.5 ， 3.75 g·L-1 ）与 A549 细胞共育不同时间（ 6 ， 24 ， 48 h ），采用流式细胞仪（ Annexin V 双标法、 DNA 含量测定和 凋亡细胞 DNA 片段原位末端检测等方法）检测凋亡细胞百分比，采用定量 Sandwich 酶免疫技术（ ELISA ）观察细胞血管内皮生长因子（ VEGF ）分泌量。 结果 3 种检测方法均表明， 中、高浓度鸦胆子油乳（ 2.5 ， 3.75 g·L-1 ）作用 6 h 以上，具有显著的诱导 A549 细胞凋亡的作用。 Annexin V 双标法和 DNA 凝胶电泳结果表明， 1.25 g·L-1 的鸦胆子油乳作用 24 ~ 48 h ，也能诱导 A549 细胞凋亡。 2.5 g · L-1 鸦胆子油乳作用 48 h 能使 A549 细胞分泌 VEGF 显著减少。 结论 鸦胆子油乳通过诱导肿瘤细胞凋亡，减少肿瘤细胞分泌 VEGF 发挥其治疗肺癌的作用。
Abstract
OBJECTIVE To investigate the effects of Brucea javanica Oil on apoptosis and vascular endothelial growth factor secretion of A549 cells and explore its mechanism of action. METHODS Percentage of apoptosis cells was evaluated by flow cytometry and VEGF secretion was determined by ELISA after cells were incubated with different concentrations of Brucea javanica oil （0.5，1.25，2.5，3.75 g·L-1）for 6, 24 or 48 h. RESULTS It was found that percentage of apoptosis cells was significantly increased after cells were incubated with Brucea javanica oil (2.5，3.75 g·L-1) for more than 6 h. The fact that Brucea javanica oil（1.25 g·L-1） induced cells to apoptosis after incubated for 24 h or 48 h was observed in Annexin V staining method and DNA gel electrophoresis, but not in other methods. Furthermore, Brucea javanica oil（2.5 g·L-1）significantly reduced VEGF secretion in A549 cells. CONCLUSION Brucea javanica oil induced apoptosis and reduced VEGF secretion of tumor cells, which might be one mechanism of Brucea javanica oil in treating pulmonary carcinosis.
关键词
鸦胆子油乳 /
A549 细胞 /
凋亡 /
血管内皮生长因子
{{custom_keyword}} /
Key words
Brucea javanica oil /
A549 cell /
apoptosis /
vascular endothelial growth factor
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] WANG F, CAO Y, LIU H Y , et al . Experimental studies on the apoptosis of HL-60 cells induced by <>Brucea javanica oil emulsion [J]. <> China J Chin Mater Med ( 中国中药杂志 ) , 2003, 28(8) : 759-762.
[2] TANG T, MENG L H, CHEN L J , et al . Reversal of multidrug resistance and inhibition of DNA topoisomerase Ⅱ by emulsion of seed oil of<> Brucea javanica [J] . <> Chin Pharmacol Bull （中国药理学通报） , 2001, 17(5): 534-539.
[3] SUN B, WU Y L, WANG S N, et al. Preliminary studies on anti- proliferative effect of <>Brucea javanica on human gastric cancer cell SGC- 7901 [J] . <> Shanghai Med J ( 上海医学 ) , 2001, 24(8): 481 - 483.
[4] LIU Y, WNAG H, FU Q J, et al . An experimental study of <>Brucea javanica oil emulsion on bladder carcinoma [J].<> <> Chin J Urol ( 中华泌尿外科杂志 ) , 2001, 22(6): 336 - 338.
[5] VERMES I, HAANEN C, REUTELINGSPERGER C P M. A novel assay for apoptosis: Flow cytometric detection of phosphatidylserine expression on early apoptoic cells using fluorescein labelled annexin V [J] . J Immunol Methods, 1995, 184(1): 39-51.
[6] MOSMANN T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays [J] . J Immunol Methods, 1983, 65(1-2): 55-63.
[7] WESSELBORG S, JANSSEN O, KABELITZ D. Induction of activation-driven death (apoptosis) in activated but not resting peripheral blood T-cell [J] . <>J Immunol, 1993, 150(10): 4338-4345.
[8] WANG J H, XU X Y, WANG Y L. Progress in As2O3 induced apoptosis of tumor cells [J] . <> Chin Pharm J （中国药学杂志） ,<> 2003, 38 (1): 1-3.
[9] QING L L , LI J S. Research development of tumor angiogenesis [J] . <>Chin J Gastroenterol Hepatal( 胃肠病学和肝病学杂志 ) , 2005, 14(2): 218-222.
[10] FERRARA N, DAVIS - SMYTH T . The biology of vascular endothelial growth factor [J] . <> Endocr Rev , 1997, 18 (1): 4-25 .
（ 收稿日期 ： 2008-09-10 ）
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
PDF(2938 KB)
