基于微流控芯片的牡丹籽粕低聚茋类化合物抗癌活性的研究
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摘要
为探究油用牡丹籽粕低聚茋类化合物对人源性乳腺癌细胞(MCF-7)活性的影响,通过设计制作一种浓度梯度微流控芯片,利用AO/PI双染鉴定细胞在不同质量浓度低聚茋类化合物诱导下的活性情况。结果表明:细胞在药液(低聚茋类化合物)诱导下,呈明显皱缩和裂解;在药液质量浓度梯度作用下,芯片每个培养腔内细胞活力不同;进行细胞计数可知,药液质量浓度为0. 5 mg/mL时,细胞活力约50%;药液质量浓度为0. 75 mg/mL时,细胞活力16. 17%;药液质量浓度达到1 mg/mL时,培养腔内几乎无正常细胞。该方法实现了对药液质量浓度梯度影响细胞活性的实时监测和观察,为微流控芯片技术与油用牡丹副产物抗癌活性研究的结合提供了一种新思路和新方法。
In order to study the inhibitory effects of oligostilbenes from peony seed meal on activity of human breast cancer cell MCF-7,by designing a novel concentration gradient microfluidic chip,AO/PI staining protocol was used to identify the activity of cells induced by different mass concentrations of oligostilbenes. The results showed that MCF-7 cells obviously crinkled and cracked under the induction of oligostilbenes. Under the action of oligostilbenes mass concentration gradient,the cells in each culture of the chip showed different activities. Cell count showed that when the mass concentration of oligostilbenes was 0. 5 mg/mL,the cell viability was about 50%. When the mass concentration was 0. 75 mg/mL,the cell viability was 16. 17%,and when the mass concentration reached 1 mg/mL,there was almost no normal cell in the culture chamber. The method realized real-time monitoring and observation of cell activity induced by drug mass concentration gradient and provided a new way of thinking and new method for the combination of microfluidic chip technology and anticancer activity of peony for oil byproducts.
In order to study the inhibitory effects of oligostilbenes from peony seed meal on activity of human breast cancer cell MCF-7,by designing a novel concentration gradient microfluidic chip,AO/PI staining protocol was used to identify the activity of cells induced by different mass concentrations of oligostilbenes. The results showed that MCF-7 cells obviously crinkled and cracked under the induction of oligostilbenes. Under the action of oligostilbenes mass concentration gradient,the cells in each culture of the chip showed different activities. Cell count showed that when the mass concentration of oligostilbenes was 0. 5 mg/mL,the cell viability was about 50%. When the mass concentration was 0. 75 mg/mL,the cell viability was 16. 17%,and when the mass concentration reached 1 mg/mL,there was almost no normal cell in the culture chamber. The method realized real-time monitoring and observation of cell activity induced by drug mass concentration gradient and provided a new way of thinking and new method for the combination of microfluidic chip technology and anticancer activity of peony for oil byproducts.
引文
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[15]SHEN S F,TIAN C,TIAN B L,et al.Spiral microchannel with ordered micro-obstacles for continuous and highly-efficient particle separation[J].Lab Chip,2017,17(21):3578-3591.
[2]牛亚琪.油用牡丹籽榨油副产物低聚茋类和单萜苷类化合物提取与纯化研究[D].河南洛阳:河南科技大学,2015.
[3]ZIGMOND S H.Ability of polymorphonuclear leukocytes to chemotaxtic factors[J].J Cell Biol,1997,75:606-616.
[4]ZICHA D,DUNN G,JONES G.Analyzing chemotaxis using the Dunn direct-viewing chamber[J].Methods Mol Biol,1997,75:449-457.
[5]HEIT B,TAVENER S,RAHARJO E,et al.An intracellular signaling hierarchy determines direction of migration in opposing chemotatic gradients[J].J Cell Biol,2002,159(1):91-102.
[6]秦鹏.微流控浓度梯度芯片的研制及其在药物筛选中的应用[D].沈阳:东北大学,2014.
[7]樊佳新,王帅,孟宪生,等.基于微流控芯片技术的荆芥诱导肺肿瘤细胞凋亡谱效关系研究[J].药学学报,2017,52(1):126-131.
[8]SOMAWEERA H,HAPUTHANTHRI S O,IBRAGUIMOVA,et al.On-chip gradient generation in 256 microfluidic cell cultures:simulation and experimental validation[J].Analyst,2015,140(15):5029-5038.
[9]KIM S,CESTELLOSBLANCO S,INOUE K,et al.Miniaturized antimicrobial susceptibility test by combining concentration gradient generation and rapid cell culturing[J].Antibiotics,2015,4(4):455-466.
[10]KILINIC D,SCHWAB J,RAMPINI S,et al.A microfluidic dual gradient generator for conducting cell-based drug combination assays[J].Integr Biol,2016,8(1):39-49.
[11]刘普,李小方,王永威,等.油用牡丹籽饼粕中低聚茋类含量测定方法研究[J].中国粮油学报,2016,31(4):138-141,146.
[12]汪耀,李颖,徐波,等.微流控芯片药物诱导细胞凋亡[J].中国科学:化学,2011,41(10):1629-1635.
[13]LIU W,LI L,WANG J C,et al.Dynamic trapping and high-throughput patterning of cells using pneumatic microstructures in an integrated microfluidic device[J].Lab Chip,2012,12(9):1702-1709.
[14]WANG L,LIU W,WANG Y,et al.Concentration of oxygen and chemical concentration gradients in a single microfluidic device for studying tumor cell-drug interactions in a dynamic hypoxia microenvironment[J].Lab Chip,2013,13(4):695-705.
[15]SHEN S F,TIAN C,TIAN B L,et al.Spiral microchannel with ordered micro-obstacles for continuous and highly-efficient particle separation[J].Lab Chip,2017,17(21):3578-3591.