新藤黄酸对人脐静脉内皮细胞凋亡的影响
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摘要
目的探讨新藤黄酸对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖和凋亡的影响。方法用不同浓度的新藤黄酸培养细胞24、48h后,采用MTT法观察新藤黄酸对细胞生长的抑制作用;倒置显微镜下观察HUVECs细胞形态变化;通过DAPI染色后倒置荧光显微镜观察新藤黄酸对细胞形态学的影响;通过Annexin V-FITC/PI双染及流式细胞术检测新藤黄酸对HUVECs细胞凋亡率的影响;Western blot检测新藤黄酸对HUVECs中凋亡相关蛋白Caspase-3的表达变化。结果 MTT检测结果显示新藤黄酸能够明显抑制HUVECs的增殖,并呈时效及量效关系;荧光显微镜观察结果显示,新藤黄酸作用HUVECs后出现大量凋亡细胞;流式细胞术结果显示,与空白对照组相比,随着新藤黄酸浓度的增加,细胞凋亡率明显增加;Western blot检测结果表明Caspase-3蛋白表达水平显著增加。结论新藤黄酸对HUVECs有明显的抑制作用,其作用机制可能是通过诱导细胞凋亡实现的。
Objective To investigate the effect of gambogenic acid(GNA)on the proliferation and apoptosis of human umbilical vein endothelial cells(HUVECs).Methods After HUVECs were cultured with different concentrations of GNA for 24 and 48 hours,MTT assay was used to observe the inhibitory effect of GNA on cell growth,and an inverted microscope was used to observe morphological changes of HUVECs.After DAPI staining,an inverted fluorescence microscope was used to observe the effect of GNA on cell morphology.Annexin Ⅴ-FITC/PI staining and flow cytometry were used to observe the effect of GNA on the apoptosis rate of HUVECs.Western blot was used to measure the effect of GNA on the expression of the apoptosis-related protein Caspase-3 in HUVECs.Results MTT assay showed that GNA significantly inhibited the proliferation of HUVECs in a time-and dose-dependent manner.Fluorescence microscopy showed the apoptosis of a large number of HUVECs after GNA treatment.Flow cytometry showed that compared with the normal control group,HUVECs treated with GNA had a significant increase in apoptosis rate,which increased with the increase in the concentration of GNA.Western blot showed a significant increase in the protein expression of Caspase-3.Conclusion GNA has a marked inhibitory effect on HUVECs,possibly by inducing cell apoptosis.
Objective To investigate the effect of gambogenic acid(GNA)on the proliferation and apoptosis of human umbilical vein endothelial cells(HUVECs).Methods After HUVECs were cultured with different concentrations of GNA for 24 and 48 hours,MTT assay was used to observe the inhibitory effect of GNA on cell growth,and an inverted microscope was used to observe morphological changes of HUVECs.After DAPI staining,an inverted fluorescence microscope was used to observe the effect of GNA on cell morphology.Annexin Ⅴ-FITC/PI staining and flow cytometry were used to observe the effect of GNA on the apoptosis rate of HUVECs.Western blot was used to measure the effect of GNA on the expression of the apoptosis-related protein Caspase-3 in HUVECs.Results MTT assay showed that GNA significantly inhibited the proliferation of HUVECs in a time-and dose-dependent manner.Fluorescence microscopy showed the apoptosis of a large number of HUVECs after GNA treatment.Flow cytometry showed that compared with the normal control group,HUVECs treated with GNA had a significant increase in apoptosis rate,which increased with the increase in the concentration of GNA.Western blot showed a significant increase in the protein expression of Caspase-3.Conclusion GNA has a marked inhibitory effect on HUVECs,possibly by inducing cell apoptosis.
引文
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[2]CHEN W Q,ZHENG R S,PETER D,et al.Cancer statistics in China,2015[J].Ca Cancer J Clin,2016,66(2):115-132.
[3]NAGHAVI M.The global burden of cancer 2013[J].JAMA Oncol,2015,1(4):505-527.
[4]玉林,文继舫,唐建武,等.病理学[M].8版.北京:人民卫生出版社,2014:88-89.
[5]李宏江,赵扬冰.抗肿瘤新生血管形成治疗实体肿瘤的研究进展[J].中国普外基础与临床杂志,2002,9(3):207-209.
[6]郭秋均,李丛煌,花宝金.解毒活血药调控肿瘤新生血管形成机制的研究进展[J].中华中医药杂志,2016(7):2699-2702.
[7]张洁,王晓慧.抗血管生成药物联合化疗在复发性卵巢癌治疗中的研究进展[J].国际妇产科学杂志,2016,43(1):70-74.
[8]国家药典委员会.中华人民共和国药典:一部[M].化学工业出版社,2000.
[9]吕归宝,杨秀贤,黄乔书.藤黄中新藤黄酸的分离及其结构[J].药学学报,1984(8):636-639.
[10]SEET R C S,WIJDICKS E F M,RABINSTEIN A A.Stroke from acute cervical internal carotid artery occlusion:treatment results and predictors of outcome[J].Archives of Neurology,2012,69(12):1615.
[11]RANIERI G,GADALETA C D,PATRUNO R,et al.A model of study for human cancer:spontaneous occurring tumors in dogs.Biological features and translation for new anticancer therapies[J].Critical Reviews in Oncology/hematology,2013,88(1):187-197.
[12]RANIERI G,PANTALEO M,PICCINNO M,et al.Tyrosine kinase inhibitors(TKIs)in human and pet tumours with special reference to breast cancer:a comparative review[J].Crit Rev Oncol Hematol,2013,88(2):293-308.
[13]何建成,王利.中医药对血管新生促进或抑制作用的研究进展[J].西安交通大学学报(医学版),2018,39(6):775-778.
[14]徐步远,潘显阁,陈崇别,等.姜黄素对Lewis肺癌小鼠血管内皮成长因子和endostatin表达的影响[J].中国临床药理学与治疗学,2017,22(2):151-154.
[15]江红,毛小刚.姜黄素对小鼠宫颈癌皮下移植瘤模型外周血和肿瘤组织MMP2、MMP9、VEGF表达的影响[J].临床和实验医学杂志,2016,15(9):832-835.
[16]李娜,高俊岩,刘敏.细胞凋亡和肿瘤的关系研究进展[J].当代医学,2009,15(16):13-14.