Wallace Charity J, Medina Scott H, ElSayed Mohamed E H
Department of Biomedical Engineering Cellular Engineering & Nano-Therapeutics Laboratory College of Engineering, University of Michigan, Ann Arbor, Michigan, 48109, USA.
Pharm Res. 2014 Apr;31(4):887-94. doi: 10.1007/s11095-013-1210-5. Epub 2013 Sep 25.
This report describes the effect of rhamnolipids (RLs), an amphiphilic biosurfactant produced by the bacterium Pseudomonas aeruginosa, on the integrity and permeability across Caco-2 cell monolayers.
We measured the trans-epithelial electrical resistance (TEER) and permeability of [(14)C]mannitol across Caco-2 cell monolayers upon incubation with 0.01-5.0% v/v RLs as a function of incubation time (30, 60, 90, and 120 min). We also studied the recovery of RL-treated Caco-2 cell monolayers upon incubation with Kaempferol, which is a natural flavonoid that promotes the assembly of the tight junctions.
TEER of Caco-2 cell monolayers incubated with 0.01-5.0% v/v RLs solution dropped to 80-28% of that of untreated cells. Decline in TEER was associated with an increase in [(14)C]mannitol permeability as a function of RLs concentration and incubation time with Caco-2 cells. Incubation of RLs-treated Caco-2 cell monolayers with normal culture medium for 48 h did not restore barrier integrity. Whereas, incubation of a RLs-treated Caco-2 cells with culture medium containing Kaempferol for 24 h restored barrier function indicated by the higher TEER and lower [(14)C]mannitol permeability values.
These results show the ability of RLs to modulate the integrity and permeability of Caco-2 cell monolayers in a concentration- and time-dependent fashion, which suggest their potential to function as a non-toxic permeation enhancer.
本报告描述了铜绿假单胞菌产生的两亲性生物表面活性剂鼠李糖脂(RLs)对Caco-2细胞单层完整性和跨膜通透性的影响。
我们在0.01 - 5.0% v/v RLs中孵育Caco-2细胞单层,测量其跨上皮电阻(TEER)以及[¹⁴C]甘露醇的通透性,并将其作为孵育时间(30、60、90和120分钟)的函数。我们还研究了用山奈酚孵育经RLs处理的Caco-2细胞单层后的恢复情况,山奈酚是一种促进紧密连接组装的天然黄酮类化合物。
用0.01 - 5.0% v/v RLs溶液孵育的Caco-2细胞单层的TEER降至未处理细胞的80% - 28%。TEER的下降与[¹⁴C]甘露醇通透性随RLs浓度和与Caco-2细胞孵育时间的增加有关。用正常培养基孵育经RLs处理的Caco-2细胞单层48小时并不能恢复屏障完整性。然而,用含山奈酚的培养基孵育经RLs处理的Caco-2细胞24小时可恢复屏障功能,表现为较高的TEER值和较低的[¹⁴C]甘露醇通透性值。
这些结果表明RLs能够以浓度和时间依赖性方式调节Caco-2细胞单层的完整性和通透性,这表明它们具有作为无毒渗透增强剂发挥作用的潜力。