Study on the Mechanism of H19/miR-93-5p/STAT3 in Regulating the Expression of Inflammatory Factors and Oxidative Stress in Microglia

Abstract

Objective: To investigate the regulatory mechanism of the H19/miR-93-5p/STAT3 pathway on the expression of inflammatory factors and oxidative stress in microglia, providing potential therapeutic targets for neuroinflammatory-related diseases. Methods: Twenty patients with chronic subdural hematoma admitted to our hospital from August 2023 to December 2024 were selected as the study subjects. BV2 microglia were extracted from their local inflammatory hyperplastic tissues for experimental analysis. The cells were randomly divided into an LPS-induced group and a normal cell control group, with 10 cases each. The LPS-induced group was further subdivided into an H19 knockdown group (n = 3) constructed by transfecting with an H19 knockdown vector; an miR-93-5p overexpression group (n = 4) formed by transfecting with an miR-93-5p mimic; and further subdivided into an H19 knockdown group (n = 3) and an miR-93-5p overexpression group (n = 4) by transfecting with an miR-93-5p mimic/inhibitor and an H19 knockdown vector. The mRNA levels of H19, miR-93-5p, and inflammatory factors (IL-1β, IL-6, TNF-a) were detected by RT-qPCR. The expression of STAT3 phosphorylation (p-STAT3), the Nrf2/HO-1 axis, and oxidative stress markers (MDA, GSH) were analyzed by Western blot. The binding relationship between STAT3 and the miR-93-5p promoter was verified by dual-luciferase assay. Results: After LPS induction, H19 expression was upregulated, miR-93-5p expression was decreased, and the levels of p-STAT3, inflammatory factors, and MDA were significantly increased (P < 0.01), while the GSH level was decreased (P < 0.05). Knockdown of H19 or overexpression of miR-93-5p could reverse these changes, inhibit p-STAT3, and activate the Nrf2/HO-1 axis, while reducing inflammatory factors and MDA (P < 0.01) and increasing GSH (P < 0.05). Dual-luciferase assay confirmed that STAT3 directly binds to the miR-93-5p promoter. Conclusion: The H19/miR-93-5p/STAT3 pathway affects the release of inflammatory factors and oxidative stress in microglia by regulating STAT3 phosphorylation and the Nrf2/HO-1 axis, providing a new strategy for the treatment of neuroinflammatory diseases.