Laboratory of General Pathology, Department of Medicine and Surgery, University of Parma, 43125 Parma, Italy.
Human Nutrition Unit, Department of Food & Drug, University of Parma, 43125 Parma, Italy.
Nutrients. 2019 Sep 21;11(10):2271. doi: 10.3390/nu11102271.
The possibility of counteracting inflammation-related barrier defects with dietary compounds such as (poly)phenols has raised much interest, but information is still scarce. We have investigated here if (+)-catechin (CAT) and procyanidin B (PB), two main dietary polyphenols, protect the barrier function of intestinal cells undergoing inflammatory stress. The cell model adopted consisted of co-cultured Caco-2 and HT29-MTX cells, while inflammatory conditions were mimicked through the incubation of epithelial cells with the conditioned medium of activated macrophages (MCM). The epithelial barrier function was monitored through trans-epithelial electrical resistance (TEER), and ROS production was assessed with dichlorofluorescein, while the expression of tight-junctional proteins and signal transduction pathways were evaluated with Western blot. The results indicated that MCM produced significant oxidative stress, the activation of NF-κB and MAPK pathways, a decrease in occludin and ZO-1 expression, and an increase in claudin-7 (CL-7) expression, while TEER was markedly lowered. Neither CAT nor PB prevented oxidative stress, transduction pathways activation, ZO-1 suppression, or TEER decrease. However, PB prevented the decrease in occludin expression and both polyphenols produced a huge increase in CL-7 abundance. It is concluded that, under the conditions adopted, CAT and PB do not prevent inflammation-dependent impairment of the epithelial barrier function of intestinal cell monolayers. However, the two compounds modify the expression of tight-junctional proteins and, in particular, markedly increase the expression of CL-7. These insights add to a better understanding of the potential biological activity of these major dietary flavan-3-ols at intestinal level.
具有抗炎作用的膳食化合物(如多酚)来对抗相关的屏障缺陷的可能性引起了广泛关注,但目前相关信息仍然匮乏。本研究旨在探究(+)-儿茶素(CAT)和原花青素 B(PB)这两种主要的膳食多酚是否能保护发生炎症应激的肠细胞的屏障功能。本研究采用共培养的 Caco-2 和 HT29-MTX 细胞作为细胞模型,通过用激活的巨噬细胞条件培养基(MCM)孵育上皮细胞来模拟炎症条件。通过跨上皮电阻(TEER)监测上皮屏障功能,用二氯荧光素评估 ROS 产生情况,用 Western blot 评估紧密连接蛋白和信号转导通路的表达情况。结果表明,MCM 产生了显著的氧化应激,NF-κB 和 MAPK 通路被激活,occludin 和 ZO-1 的表达减少,claudin-7(CL-7)的表达增加,同时 TEER 明显降低。CAT 和 PB 均不能预防氧化应激、转导通路激活、ZO-1 抑制或 TEER 降低。然而,PB 能防止 occludin 表达的减少,两种多酚都能显著增加 CL-7 的丰度。综上所述,在所采用的条件下,CAT 和 PB 不能预防炎症导致的肠细胞单层上皮屏障功能障碍。然而,这两种化合物可以调节紧密连接蛋白的表达,特别是显著增加 CL-7 的表达。这些研究结果有助于更好地了解这两种主要膳食黄烷-3-醇在肠道水平的潜在生物学活性。