Objective To investigate the mechanism of total triterpenes from fruits of Chaenomeles speciosa (CST) in alleviating N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and (NIG)-induced human gastric mucosal epithelial cells damage based on microRNA-210 (miR-210)/nuclear factor-κB (NF-κB)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome signaling pathway. Methods MNNG and NIG induced GES-1 cells model were constructed, and then GES-1 cells were treated with CST and/or miR-210 mimic, miR-210 inhibitor, NF-κB siRNA, NLRP3 siRNA. MTT assay was used to detect cell viability, immunofluorescence was utilized to measure reactive oxygen species (ROS) level, flow cytometry was used to test pyroptosis, colorimetric and ELISA methods were utilized to measure the levels of interleukin-4 (IL-4), IL-1β, IL-6, IL-10, IL-18, lactate dehydrogenase (LDH), and tumor necrosis factor-α (TNF-α) in cell supernatant. The colorimetric method was used to determine the levels of catalase (CAT), glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in cells. The dual luciferase reporter assay was employed to verify the targeted intervention between miR-210 and NF-κB. Immunofluorescence was utilized to detect the NF-κB p65 nuclear translocation, the co-localization of NLRP3, apoptosis related spot like protein (ASC) and pro-cystein-asparate protease-1 (pro-Caspase-1). qRT-PCR was used to detect the mRNA expression levels of miR-210, NF-κB, NLRP3. Western blotting was utilized to determine the protein expression levels of cytoplasmic NF-κB, nuclear NF-κB, NIMA-related kinase 7 (NEK7), thioredoxin-interacting protein (TXNIP), NLRP3, ASC, Caspase-1, pro-Caspase-1, gasdermin D (GSDMD), N-terminal domain of GSDMD (GSDMD-N), pro-IL-18 and pro-IL-1β. Results miR-210 was a pro-inflammatory/pyroptosis factor, the dual luciferase reporter assay confirmed that it could bind to 3’-UTR in mRNA of NF-κB. Compared with model group, CST significantly inhibited pyroptosis and LDH release in MNNG and NIG induced GES-1 cells (P < 0.01), reduced levels of IL-1β, IL-6, IL-18, TNF-α in supernatant and ROS, MPO, MDA in cells (P < 0.01), elevated levels of IL-4, IL-10 in supernatant and activities of CAT, GSH, SOD, T-AOC in cells (P < 0.01), suppressed the NF-κB p65 nuclear translocation and the co-localization of NLRP3, ASC and pro-Caspase-1 (P < 0.01), downregulated the mRNA expressions of miR-210, NF-κB, NLRP3 and the protein expressions of nuclear NF-κB p65, TXNIP, NEK7, NLRP3, ASC, pro-Caspase-1, Caspase-1, GSDMD, GSDMD-N, pro-IL-1β, pro-IL-18 (P < 0.01), upregulated the cytoplasmic NF-κB p65 protein expression (P < 0.01). The combination of CST with miR-210 inhibitor, NF-κB siRNA and NLRP3 siRNA transfection could further improve the above indicators (P < 0.01). Conclusion CST has a good preventive and therapeutic effect on MNNG and NIG-induced GES-1 cell injury, and its mechanism is closely related to inhibiting the abnormal elevation of miR-210 and activation of NF-κB/NLRP3 inflammasome signaling pathway, alleviating oxidative stress and subsequent suppressing pyroptosis.

total triterpenes from fruits of Chaenomeles speciosa  /  inflammatory response  /  oxidative stress  /  pyroptosis  /  miR-210/NF-κB/NLRP3 inflammasome signaling pathway  /  chronic atrophic gastritis