Abstract
Glycative stress, caused by the non-enzymatic reaction of reactive carbonyl compounds such as methylglyoxal (MGO) and glyoxal (GO) with proteins, contributes to ageing and lifestyle-related diseases through mechanisms distinct from, yet closely linked to, oxidative stress. This study investigated whether Yoda1, a selective Piezo1 agonist, and Manda Koso, a naturally fermented food supplement, protect against glycative stress in rat intestinal epithelial (RIE) cells through direct chemical activity or intracellular signalling.
Direct anti-glycation activity was evaluated using DAN-HPLC aldehyde-trapping and HSA–glucose glycation-inhibition assays. Yoda1 showed no detectable aldehyde-trapping activity toward 3-deoxyglucosone (3DG), GO, or MGO, nor glycation- inhibitory activity toward fluorescent AGEs (FAGEs), 3DG, GO, or MGO, indicating that it possesses no intrinsic carbonyl- scavenging activity. In contrast, Manda Koso markedly inhibited AGE formation and carbonyl intermediate generation in a dose-dependent manner, with modest trapping of 3DG and MGO.
Cell-based analyses demonstrated that non-cytotoxic Yoda1 (1 μM) induced rapid cytoplasmic Ca² + influx, followed by mitochondrial Ca ² + uptake and a low-level generation of mitochondrial reactive oxygen species (ROS), resulting in a significant elevation of antioxidant capacity, measured by the Singlet Oxygen Absorption Capacity (SOAC) assay. These effects were completely abolished by the Piezo1 inhibitors Dooku1 and GsMTx4, confirming a Piezo1-dependent mechanism. Manda Koso elicited a slower Ca² + response and only partial inhibition by Piezo1 blockers, indicating the coexistence of signalling-dependent and direct chemical mechanisms.
These findings demonstrate that Yoda1 enhances antioxidant capacity exclusively through the intracellular Piezo1– Ca² +–low- level mitochondrial ROS–hormesis pathway, culminating in increased endogenous antioxidant capacity, without direct anti- glycation activity, whereas Manda Koso acts through complementary signalling and direct anti-glycation mechanisms. Piezo1- mediated mechanotransduction may therefore represent a novel signalling pathway linking hormetic antioxidant defence with glycative stress biology in intestinal epithelial cells.
