Effect of Indoxyl Sulfate on the Repair and Intactness of Intestinal Epithelial Cells: Role of Reactive Oxygen Species’ Release

Abstract

Chronic kidney disease (CKD) is characterized by an oxidative stress status, driving some CKD-associated complications, even at the gastrointestinal level. Indoxyl Sulfate (IS) is a protein-bound uremic toxin, poorly eliminated by dialysis. This toxin is able to affect the intestinal system, but its molecular mechanism/s in intestinal epithelial cells (IECs) remain poorly understood. This study’s aim was to evaluate the effect of IS (31.2–250 µM) on oxidative stress in IEC-6 cells and on the intactness of IECs monolayers. Our results indicated that IS enhanced oxidative cell damage by inducing reactive oxygen species (ROS) release, reducing the antioxidant response and affecting Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) nuclear translocation as well its related antioxidant enzymes. In the wound healing assay model, IS reduced IEC-6 migration, slightly impaired actin cytoskeleton rearrangement; this effect was associated with connexin 43 alteration. Moreover, we reported the effect of CKD patients’ sera in IEC-6 cells. Our results indicated that patient sera induced ROS release in IEC-6 cells directly related to IS sera content and this effect was reduced by AST-120 serum treatment. Results highlighted the effect of IS in inducing oxidative stress in IECs and in impairing the intactness of the IECs cell monolayer, thus significantly contributing to CKD-associated intestinal alterations.

Bioactive Polyphenols from Pomegranate Juice Reduce 5-Fluorouracil-Induced Intestinal Mucositis in Intestinal Epithelial Cells

“…IEC-6 cells were subsequently collected, washed with PBS, and incubated in PBS with H 2 DCF-DA (10 µM). After an incubation of 15 min at 37 °C cell fluorescence was evaluated with a fluorescence-activated cell sorter (FACSscan; Becton Dickinson, Franklin Lakes, NJ, USA), and was analyzed by CellQuest software (version 4; Becton Dickinson, Milan, Italy) [ 28 ].…”

“…The analytical platform developed led to detection of a higher number of tannins that were eluted in the middle and final part of the 2D plot. Several ellagic and galloyl glycosides were found, which presented a similar MS/MS fragmentation resulting from a hexose [M-H-162] - (28,29,30,32,33,34,35,38,42,48). In addition to ellagic acid hexoside (m/z 463) further monoglycosylated ellagic acid derivatives were observed, such as an ellagic acid-pentoside (m/z 433; peak 26) and deoxyhexoside (m/z 447; peaks 23, 24, 27), all of them showing the typical fragments of ellagic acid at m/z 300.…”

Inflammation and Oxidative Stress in Chronic Kidney Disease—Potential Therapeutic Role of Minerals, Vitamins and Plant-Derived Metabolites

“…Many studies have reported the effects of uremic toxins from the intestine. These gut-derived uremic toxins have been shown to play a pivotal role in affecting intestinal homeostasis, as well as inducing inflammation and oxidative stress in the systemic circulation [45,46]. Uremic toxins have been positively associated with NF-κB expression and exhibit positive correlations with CRP levels, iNOS and COX-2 expression [43,47].…”

“…Several natural-derived agents have been used in Chinese herbal medicine, including flavonoids (or bioflavonoids), which form an important group of secondary plant metabolites ( Table 2). Many studies have provided evidence supporting the importance of nutrition during CKD; in particular, the role of nutrition in regulating the levels of gut-derived uremic toxins has been demonstrated to be beneficial in CKD patients [45,[123][124][125][126].…”

Citrus sinensis and Vitis vinifera Protect Cardiomyocytes from Doxorubicin-Induced Oxidative Stress: Evaluation of Onconutraceutical Potential of Vegetable Smoothies

“…In the presence of intracellular ROS, H 2 DCF is rapidly oxidized to the highly fluorescent 2 ,7 -dichlorofluorescein (DCF). After cell treatment, previously reported, cells were collected, washed twice with phosphate buffer saline (PBS), and then incubated in PBS containing H 2 DCF-DA (10 µM) at 37 • C. After 15 min, fluorescence was evaluated using a fluorescence-activated cell sorting (BD FacsCalibur, Milan, Italy) and elaborated with Cell Quest software, as previously reported [20].…”