The role of Mir-365-5p in regulating NF2-Mediated apoptosis and ROS production in hypoxic cardiomyoblasts

Background: Prolonged hypoxia is known to induce reactive oxygen species (ROS) accumulation, leading to cellular damage, particularly in cardiac cells. MicroRNAs (miRNAs) have emerged as key regulators in cellular responses to hypoxia. This study investigated the role of miR-365-5p in hypoxia-induced injury in H9c2 cardiomyoblasts and explored the potential protective effects of bioactive compounds derived from the Jing Si herbal tea. Methods and results: Microarray analysis was employed to identify differentially expressed miRNAs in H9c2 cells subjected to 24-hour hypoxic conditions. Among several candidates, mir-365-5p was found to be significantly downregulated, correlating with the upregulation of neurofibromin 2 (NF2), a known regulator of apoptotic signaling pathways. Functional studies using a mir-365-5p mimic revealed that NF2 expression was suppressed, ROS accumulation was decreased, and intrinsic apoptosis was attenuated. Similarly, NF2 knockdown via siRNA recapitulated the protective effects of the mir-365-5p mimic under hypoxic conditions. In parallel, high-performance liquid chromatography (HPLC) was utilized to isolate three individual compounds from Jing Si herbal tea. Treatment with these compounds prevented the hypoxia-induced downregulation of mir-365-5p, suggesting a mechanism for their cardioprotective action. Conclusions: The mir-365-5p served as a crucial modulator of hypoxia-induced oxidative stress and apoptosis in H9c2 cardiomyoblasts through the regulation of NF2. Compounds derived from Jing Si herbal tea may exert cardioprotective effects by sustaining mir-365-5p expression under hypoxic conditions. These findings highlight the therapeutic potential of targeting the mir-365-5p/NF2 axis in cardiac injury.