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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(Te A)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Te A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AM E的Papp大小与毒素浓度和转运时间有关,AOH、AME和Te A为易于吸收或中等易吸收的毒素。
Objective To investigate the transmembrane transport and absorption characteristics of Caco-2 cell model by using Caco-2 cell model. Methods Caco-2 cells were seeded on Transwell plates for 21 days. The cells were incubated with TEER, ALP, Papp, Morphology and other indicators to evaluate the construction of Caco-2 cell model. The concentrations of carbendazim (AOH), carbendim or monomethylether (AME), carbendazim, Te A) solution transport and absorption. Results The TEER of cultured Caco-2 cells cultured for 21 days was more than (300 ± 15) Ω · cm2, and there was significant difference in ALP activity between the apical side and basement membrane (P <0.01). Transmission electron microscopy shows clear microvilli and tight junctions between cells. The Papp range of AOH, AME and Te A was (1 ~ 10) × 10-5cm / s, and the range of absorbance was 1.5% -63.0% at different concentrations and transit times. Conclusion The established model formed a complete and compact cell monolayer. The Papp size of AOH and AM E is related to the toxin concentration and transit time, and AOH, AME and Te A are toxins that are readily or moderately absorbed.