Network pharmacology and experimental validation reveal inhibition of the JAK2/STAT3 pathway by hydrogen sulfide in diabetic cardiomyopathy

Hydrogen sulfide (H 2 S) is well-known for its role as a gasotransmitter with cardioprotective properties, particularly in the context of diabetic cardiomyopathy (DCM). However, its molecular mechanisms are not completely defined. The current study outlines how H 2 S alleviates DCM through combination of network pharmacology and experimental validations. A spontaneous type 2 diabetes-associated DCM model was developed using db/db mice which were treated with the H₂S donor sodium hydrosulfide (NaHS) for 8 weeks. The functional, histological, and molecular evaluations were broadly analyzed for the cardiac protective effects of H 2 S. NaHS treatment significantly attenuated myocardial fibrosis and inflammation in db/db mice, which thereby leading to a pronounced improvement in cardiac function. Network pharmacology analyses identified 121 overlapping targets between H₂S and DCM, which were enriched in various signaling pathways mainly associated with inflammation and ECM remodeling. Topological analysis indicated that the JAK2/STAT3 signaling pathway serves as a crucial mediator, with STAT3 identified as a central hub among various analytical modules. According to the experimental results, the JAK2/STAT3 signaling was significantly activated in both diabetic hearts and high-glucose stimulated cardiac fibroblasts. In contrast, H₂S treatment significantly inhibited this activation. More importantly, the pharmacological reactivation of STAT3 largely reversed the protective effects of H₂S both in vivo and in vitro. Taken together, our results demonstrate that H₂S alleviates DCM via inhibiting inflammatory and fibrotic responses that associated with JAK2/STAT3 signaling pathway. This study provides a mechanistic insight into H₂S-mediated cardioprotection and it highlights the JAK2/STAT3 signaling as a potential therapeutic target in DCM.