Zishen Huoxue Decoction alleviates myocardial ischemia-reperfusion injury through dysregulation of endoplasmic reticulum-mitochondria homeostasis mediated by DUSP1-NDUFS4

Zishen Huoxue (ZSHX) Decoction can ameliorate myocardial ischaemia by regulating the mitochondrial quality control network. However, the identification of new molecular targets is necessary for ZSHX’s control of mitochondrial protein homeostasis and metabolic activities. Utilizing animal and cellular models with NDUFS4CKO or DUSP1CKO, along with single-cell sequencing, metabolomics, network pharmacology, and in vivo/in vitro interventions, the study found that ischemia-reperfusion (I/R) injury triggers endoplasmic reticulum stress and mitochondrial metabolic reprogramming, accompanied by downregulation of DUSP1 and NDUFS4. Network pharmacology suggested ZSHX’s role in regulating mitochondrial activity during inflammatory damage, while metabolomics confirmed that ZSHX alters metabolite composition and expression in I/R-affected tissues. Single-cell sequencing further linked I/R to disrupted mitochondrial energy metabolism and cell death, and in vitro experiments demonstrated that ZSHX preserves mitochondrial proteostasis, inhibits endoplasmic reticulum stress, restores calcium balance, upregulates DUSP1/NDUFS4 expression, and controls metabolic reprogramming to reduce myocardial inflammatory injury. Kaempferol, the primary active component of ZSHX, drives these protective effects by enhancing DUSP1/NDUFS4 expression, thereby preventing endoplasmic reticulum stress and inflammatory bursts, preserving mitochondrial function, and re-encoding mitochondrial metabolic processes post-I/R injury.