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山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (6): 18-22.doi: 10.6040/j.issn.1671-7554.0.2015.245

• 基础医学 • 上一篇    下一篇

交链孢毒素在Caco-2细胞中的转运与吸收

赵凯1,2, 韩春卉2, 林肖惠2, 江涛2, 杨大进2, 宋敬东3, 李凤琴2   

  1. 1. 中国疾病预防控制中心营养与健康所, 北京 100050;
    2. 国家食品安全风险评估中心卫生部食品安全风险评估重点实验室, 北京 100021;
    3. 中国疾病预防控制中心病毒病预防控制所, 北京 102206
  • 收稿日期:2015-03-07 修回日期:2015-04-16 出版日期:2015-06-10 发布日期:2015-06-10
  • 通讯作者: 李凤琴。 E-mail:lifengqin@cfsa.net.cn E-mail:lifengqin@cfsa.net.cn
  • 基金资助:
    国家高技术研究发展(863)计划(2012AA101603)

Transportation and absorption of Alternaria mycotoxins in Caco-2 cells

ZHAO Kai1,2, HAN Chunhui2, LIN Xiaohui2, JIANG Tao2, YANG Dajin2, SONG Jingdong3, LI Fengqin2   

  1. 1. National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention, Beijing 100050, China;
    2. Key Laboratory of Food Safety Risk Assessment of Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China;
    3. Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
  • Received:2015-03-07 Revised:2015-04-16 Online:2015-06-10 Published:2015-06-10

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摘要： 目的 利用构建的结肠腺癌细胞系(Caco-2)细胞模型,探讨交链孢毒素经Caco-2细胞模型的跨膜转运与吸收特征。方法 将Caco-2细胞接种于Transwell培养板的聚碳酯膜上培养21 d,用跨上皮细胞电阻(TEER)、碱性磷酸酶(ALP)活性、荧光素表观渗透系数(Papp)和细胞形态学等指标评价所构建的Caco-2细胞模型。通过测定细胞单层顶侧和基底膜侧交链孢毒素的含量,研究不同浓度和转运时间交链孢酚(AOH)、交链孢酚单甲醚(AME)、交链孢菌酮酸(TeA)溶液的转运与吸收。结果 培养至第21天的Caco-2细胞其TEER超过(300±15)Ω·cm2,且细胞顶侧和基底膜侧ALP活性差异有统计学意义(P < 0.01)。透射电镜下可见清晰的微绒毛和细胞间的紧密连接。AOH、AME和TeA的Papp范围为(1~10)×10-5cm/s,不同浓度和转运时间的毒素吸收率范围为1.5%~63.0%。结论 建立的模型形成了完整且紧密的细胞单层。AOH和AME的Papp大小与毒素浓度和转运时间有关,AOH、AME和TeA为易于吸收或中等易吸收的毒素。

关键词: 转运, 模型, 交链孢毒素, 吸收, Caco-2细胞

Abstract: Objective To develop and evaluate the Caco-2 cell model and to explore the transportation and absorption of 3 Alternaria mycotoxins. Methods Caco-2 cell model was established by inoculating Caco-2 cell on the polycarbonate membrane attached to the transwell plates and evaluated on the basis of transepithelium electrical resistance (TEER), activity of alkaline phosphatase (ALP), apparent permeability coefficient (Papp) of fluorescein and cytomorphology. Transportation and absorption of 3 Alternaria toxins including alternariol (AOH), alternariol methyl ether (AME) and tenuazonic acid (TeA) in different concentrations and at different time points were studied by determining the difference in toxin concentrations between apical side and basolateral side of the cell line. Results The TEER exceeded 300±15 Ω·cm2 and ALP activity was statistically different (P<0.01) between apical side and basolateral side after 21 days' incubation. Clear microvilli and tight intercellular junctions were observed with transmission electron microscope. Papps of AOH, AME and TeA varied between 1×10-5cm/s and 10×10-5cm/s. Transportation and absorptionof 3 Alternaria toxins at different concentrations ranged from 1.5% to 63.0%. Conclusion Intact and tight cell model was developed for Alternaria toxins transportation and absorption. AOH, AME and TeA were toxins easily or moderately absorbed by human beings.

Key words: Alternaria mycotoxins, Absorption, Caco-2 cell, Transportation, Model

中图分类号: 

  • R155.5
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