基于阵列微流控细胞芯片的植物组分抗氧化活性分析

引用本文: 张潇丹, 徐溢, 张涛, 吕君江. 基于阵列微流控细胞芯片的植物组分抗氧化活性分析[J]. 分析化学, 2016, 44(4): 604-609. doi: 10.11895/j.issn.0253-3820.150596 shu

Citation: ZHANG Xiao-Dan, XU Yi, ZHANG Tao, LÜ Jun-Jiang. Assessing Plant Antioxidants by Cellular Antioxidant Activity Assay Based on Microfluidic Cell Chip with Arrayed Microchannels[J]. Chinese Journal of Analytical Chemistry, 2016, 44(4): 604-609. doi: 10.11895/j.issn.0253-3820.150596 shu

基于阵列微流控细胞芯片的植物组分抗氧化活性分析

1.
¹ 重庆大学化学化工学院, 重庆 400030;
2.
² 新型微纳器件与系统技术重点学科实验室, 重庆 400030;
3.
³ 微纳系统及新材料技术国际研发中心, 重庆 400030

通讯作者: 徐溢

基金项目:
本文系国家自然科学基金项目(No.21375156) (No.21375156)

国家高技术研究发展计划(863)项目(No.2015AA021104) (863)项目(No.2015AA021104)

中央高校基本科研业务费专项重点项目(No.106112015CDJZR225512) (No.106112015CDJZR225512)

重庆市科技攻关项目(No.cstc2015jcsf80001)资助 (No.cstc2015jcsf80001)

摘要: 设计并制作了一种集成有8个重复6×6细胞培养单元的阵列微流控细胞芯片,以实现细胞培养和系列植物组分的细胞抗氧化活性(Cellular antioxidant activity,CAA)分析。芯片主要包含聚二甲基硅烷盖片、288个圆形培养腔体,48个独立平行通道的玻璃基底层,一次可完成8个样本的6个浓度筛选,并可在酶标仪上实现测试。槲皮素、芦丁和山奈酚等植物组分与芯片上培养的细胞作用24 h,细胞存活率大于90%。以芯片上培养的人肝癌细胞HepG2为细胞载体,以2',7'-二氯荧光素-乙酰乙酸酯(2',7'-Dichlorofluorescin diacetate,DCFH-DA)为荧光探针,采用2,2'-偶氮二异丁基脒二盐酸盐(2,2'-azobis(2-amidinopropane) dihydrochloride,ABAP)为细胞内活性氧(Reactive oxygen species,ROS)引发剂,测得槲皮素、芦丁、山奈酚等植物组分的CAA unit分别为71.42±0.19、74.31±0.36和69.92±0.09(x±s,n=3),IC₅₀分别为(7.20±0.06) μmol/L,(52.06±0.14) μmol/L,(32.55±0.03) μmol/L(x±s,n=3)。

关键词:

English

Assessing Plant Antioxidants by Cellular Antioxidant Activity Assay Based on Microfluidic Cell Chip with Arrayed Microchannels

1.
¹ College of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400030, China;
2.
² Key Disciplines Lab of Novel Micro-nano Devices and System Technology, Chongqing 400030, China;
3.
³ International R & D center of Micro-nano Systems and New Materials Technology, Chongqing 400030, China

Corresponding author: 徐溢

Received Date: 27 July 2015
Fund Project:
本文系国家自然科学基金项目(No.21375156)

国家高技术研究发展计划(863)项目(No.2015AA021104)

中央高校基本科研业务费专项重点项目(No.106112015CDJZR225512)

重庆市科技攻关项目(No.cstc2015jcsf80001)资助

Abstract: An integrated microfluidic chip with arrayed micro channels that consisted of eight repeat arrayed 6×6 cell culture chamber was designed and fabricated. The analytical microsystem combined with designed microchip, measuring device and environmental control unit was established for cell culture and parallel cellular antioxidant activity(CAA) analysis of plant antioxidants. The microfluidic chip included a polydimethylsiloxane(PDMS) cover and glass substrate that consisted of two hundreds and eighty-eight round cell culture microchambers and forty-eight independent parallel array channels. Eight groups of different samples with six different concentrations could be investigated simultaneously with multimode reader in one test. HepG2 cells were successfully cultured on the microchip. Moreover, the viability percentage of the HepG2 cells exposed to these plant antioxidants solutions at different concentrations for 24 h was higher than 90%. With 2', 7'-Dichlorofluorescin diacetate(DCFH-DA) as a fluorescence probe, 2, 2'-azobis(2-amidinopropane) dihydrochloride(ABAP) as the initiator of intracellular reactive oxygen species(ROS), we tested the inhibitory effect of several plant antioxidants such as quercetin, rutin and kaempferol on free radicals. The CAA units were calculated by the data measured from cellular morphology and fluorescence intensity over time. It was shown that the CAA units of quercetin, rutin and kaempferol were 71.42±0.19, 74.31±0.36 and 69.92±0.09(x±s, n=3), while the calculated IC₅₀ were(7.20±0.06) μmol/L,(52.06±0.14) μmol/L and(32.55±0.03) μmol/L(x±s, n=3), respectively.

Key words:

Microfluidic cell chip with arrayed microchannels
/ Cellular antioxidant activity
/ High throughput
/ Screening

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沈阳化工大学材料科学与工程学院沈阳 110142

基于阵列微流控细胞芯片的植物组分抗氧化活性分析

通讯作者: 徐溢

English

Assessing Plant Antioxidants by Cellular Antioxidant Activity Assay Based on Microfluidic Cell Chip with Arrayed Microchannels

Corresponding author: 徐溢

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于阵列微流控细胞芯片的植物组分抗氧化活性分析

通讯作者: 徐溢

English

Assessing Plant Antioxidants by Cellular Antioxidant Activity Assay Based on Microfluidic Cell Chip with Arrayed Microchannels

Corresponding author: 徐溢

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

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CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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