基于微流控芯片的细胞迁移模型及其在黄芩苷药效学研究上的应用
A cell migration model based on microfluidics and its application for baicalin pharmacodynamics research
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摘要: 基于微流控芯片技术建立一种体外细胞迁移模型，并将其用于黄芩苷药效的研究。设计与制作一种集成有微阀的微流控芯片；LIVE/DEAD染色试剂盒检测芯片中细胞活力；通过荧光素钠对芯片进行药物浓度梯度表征；DiI、DiO对细胞示踪染色观察微阀对细胞的隔离作用；每12小时显微镜拍照观察细胞的迁移状态并计算细胞迁移速率；ELISA法测定芯片细胞上清液中MMP-2、MMP-9和E-cadherin蛋白含量变化。实验结果表明，芯片中细胞活力接近100%；芯片可产生稳定的浓度梯度，且微阀对细胞有良好的隔离作用；黄芩苷对肝癌细胞HepG2的迁移具有显著抑制作用，并可显著降低MMP-2和MMP-9蛋白的分泌量。该研究为微流控芯片技术在中医药领域中的应用提供了一种新思路和新方法。
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关键词:
- 微流控芯片技术 /
- 细胞迁移 /
- 黄芩苷 /
- 人肝癌细胞HepG2 /
- MMP-2 /
- MMP-9 /
- E-cadherin
Abstract: An in-vitro model of cell migration based on microfluidic technology was established to study the pharmacodynamics of baicalin. Fabrication and design of a microfluidic chip that integrated microvalve. The cells viability on the chip was detected by LIVE/DEAD cell staining kit. Fluorescein sodium was used to measure the drug concentration gradient of the drug on the chip. DiI and DiO labeled tumor cells were added into chip and facilitated the observation over the function of the microvalve to segregate the cells. The migration of cells was observed and calculated every 12 h. ELISA assay was used to measure the concentration of MMP-2, MMP-9 as well as E-cadherin in the culture medium. These finding indicated that the viability of cells was upto 100%. The chip was able to produce the stable concentration gradient and the microvalve also functioned to segregate the cells very well. The baicalin exhibited inhibitory effect on the migration of human hepatoma HepG2 cells via decreasing the secretion MMP-2 and MMP-9 proteins(P< 0. 05). Taken together, this study provided novel application of microfluidic technology in the Chinese medicine field.-
Keywords:
- microfluidic technology /
- cell migration /
- baicalin /
- human hepatoma HepG2 cells /
- MMP-2 /
- MMP-9 /
- E-cadherin
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