TRPV4-linked oxidative–lysosomal stress in myocardial ischemia/reperfusion: disease-matched transcriptomics and functional analysis in OGD/R cardiomyoblasts

Background Myocardial ischemia/reperfusion (MI/R) injury involves oxidative stress, lysosomal dysfunction, and altered autophagy. TRPV4 may link Ca 2+ dysregulation to these processes, but its role in lysosomal membrane permeabilization (LMP) and autophagic impairment remains unclear. Methods Three disease-matched mouse-heart GEO cohorts (GSE281940, GSE130217, and GSE160516) were analyzed using limma-based differential expression, prespecified oxidative, lysosomal, and autophagy-related gene sets, cross-cohort comparisons, and within-cohort correlations. H9C2 cardiomyoblasts subjected to OGD/R were used to examine intracellular Ca 2+ , ROS, LMP, autophagy-related changes, inflammatory cytokine release, and cell injury after pharmacological modulation of TRPV4 and ROS. Results All three cohorts showed clear MI/R-associated transcriptional changes. Hmox1 was consistently increased across all five prespecified contrasts, whereas Trpv4 showed modest, time-dependent changes without reaching FDR-adjusted significance. Trpv4 was positively correlated with Map1lc3b in all three cohorts (Spearman ρ = 0.73–0.83, nominal P < 0.05), while associations with oxidative and lysosomal genes were more variable. In H9C2 cardiomyoblasts, OGD/R increased TRPV4 mRNA expression, intracellular Ca 2+ , and ROS levels. TRPV4 inhibition reduced ROS accumulation, attenuated LMP-related changes, and decreased LC3B and p62 accumulation, whereas TRPV4 activation produced the opposite effects. NAC attenuated TRPV4 activation-induced ROS and lysosomal injury, while t-BHP partially reversed the effects of TRPV4 inhibition. OGD/R also increased IL-1β, IL-6, and TNF-α release, which was reduced by TRPV4 inhibition. Conclusion The transcriptomic data indicate a context-dependent TRPV4 response within a conserved MI/R stress background. The H9C2 cardiomyoblasts experiments support the involvement of the TRPV4–ROS–LMP pathway in lysosomal injury, impaired autophagic degradation, inflammatory responses, and OGD/R-induced cardiomyoblast injury.