摘要
目的:探讨白术内酯Ⅱ对人大肠癌Lovo细胞的抑制作用及其机制。方法:体外培养人大肠癌Lovo细胞,采用不同浓度的白术内酯Ⅱ分别作用于细胞,噻唑蓝(MTT)比色法检测其对细胞生长抑制率,流式细胞术(FCM)检测白术内酯Ⅱ对Lovo细胞凋亡的影响,蛋白免疫印迹法(Western blot)检测白术内酯Ⅱ对大肠癌细胞聚腺苷酸二磷酸核糖转移酶-1(poly ADPribose polymerase1,PARP1),含半胱氨酸的天冬氨酸蛋白水解酶-3(cysteinyl aspartate specific proteinase-3,Caspase-3)蛋白表达水平的影响。结果:与空白组比较,白术内酯Ⅱ质量浓度达到150 mg·L^(-1)时可明显抑制Lovo细胞增殖(P<0.05),随着白术内酯Ⅱ浓度的增高,细胞存活率逐渐下降,当质量浓度超过300 mg·L^(-1)时,细胞存活率降到半数以下;白术内酯Ⅱ(300 mg·L^(-1))可明显促进Lovo细胞的凋亡(P<0.01);白术内酯Ⅱ100 mg·L^(-1)可使PARP1和Caspase-3开始发生剪切,随着药物质量浓度的增加,其剪切作用明显增强。白术内酯Ⅱ可抑制大肠癌Lovo细胞的生长和増殖,并能调节PARP1,Caspase-3蛋白表达,促进细胞凋亡。结论:白术内酯Ⅱ能显著抑制Lovo细胞的增殖并诱导其发生凋亡,其分子机制与促进PARP1的剪切,激活Caspase-3的表达相关。
Objective： To investigate the inhibitory effect of atractylenolide Ⅱ on Lovo cells and its mechanism. Method： Lovo cells were cultured in vitro, and treated with respectively atractylenolide Ⅱ at different concentrations to incubate for 24 h. The proliferation ability of Lovo cells was detected by 3- （4, 5-dimethyl-2- thiazolyl）-2, 5-diphenyl-2-H-tetrazolium bromide （MTT） assay. The apoptosis of Lovo cells impacted by atractylenolide 1/ were measured by flow cytometry （FCM） , and the effect of atractylenolide Ⅱ on the expression of poly ADP-ribose polymerasel （ PARP1 ） and cysteinyl aspartate specific proteinase-3 （Caspase-3） were detected by the Western blot. Result： Compared with the blank control group, at the dose of 150 mg·L^-1, atractylenolide Ⅱ significantly inhibited the proliferation of Lovo cells （P 〈 0.05 ）. With the increase of the concentration of atractylenolide Ⅱ , cell survival rate was gradually decreasing. At the concentration of more than 300 mg·L^-1, cell survival rate dropped to less than half. High-dose （300 mg·L^-1） atractylenolide Ⅱ can significantly promote the apoptosis of Lovo cells （P 〈0.01 ）. At the dose of 100 mg·L^-1 , atraetylenolide 11 can cause shear in PARP1 and Caspase-3. With the increase of drug concentration, the shearing effect was significantly enhanced. Atractylenolide Ⅱ can inhibit the growth and proliferation of Lovo cells, regulate the expression of PARP1 and Caspase-3, and then promote the apoptosis of cells. Conclusion： Atraetylenolide Ⅱ can significantly inhibit the proliferation of Lovo cells and induce their apoptosis, its molecular mechanism may be related to the regulation of the expression of PARP1 and Caspase-3.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第5期157-161,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
江苏省“六大人才高峰”第十二批高层次人才选拔培养计划项目(WSW-069)
江苏省第四期“333工程”科研计划项目(24)
南京市医学科技发展项目(ZKX15040)
