Mechanism of Yangxin Dawa Yimixike Mi Gao for myocardial ischemia/reperfusion injury: an integrative study of network pharmacology and and validation.

UHPLC-Q-Orbitrap-HRMS fingerprinting of YXDW identified 137 bioactive constituents with high batch-to-batch consistency.

• 137 total bioactive constituents were identified using ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS)
• 23 compounds demonstrated high oral bioavailability (OB ≥ 56%)
• Batch-to-batch similarity values were greater than 0.87, indicating high formulation consistency

Network pharmacology and Mendelian randomization analyses identified 13 key targets associated with myocardial I/R injury.

• Key targets identified included C-X-C motif chemokine receptor 1 (CXCR1), mitogen-activated protein kinase kinase 1 (MAP2K1), and poly (ADP-ribose) polymerase 1 (PARP1)
• These targets were predicted to be involved in regulation of apoptosis, oxidative stress, and inflammatory responses
• Mendelian randomization was used to validate network pharmacology predictions, adding causal inference support to target identification
• Gene ontology and KEGG enrichment analyses together with molecular docking were performed to identify core regulatory pathways

In vivo YXDW administration alleviated I/R-induced myocardial injury as evidenced by reduced serum injury markers and improved cardiac function.

• YXDW treatment reduced serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels in Sprague-Dawley rat I/R models
• YXDW attenuated myocardial apoptosis as assessed by histopathological examination
• Improved left ventricular function was demonstrated by echocardiography
• Inflammatory markers including C-reactive protein, interleukin-6, and tumor necrosis factor-alpha were measured as biochemical endpoints

YXDW dose-dependently downregulated CXCR1, MAP2K1, and PARP1 expression at both protein and mRNA levels in myocardial tissues and H/R-injured H9c2 cardiomyocytes.

• Downregulation of target gene expression was observed both in vivo (rat myocardial I/R model) and in vitro (H9c2 cardiomyocyte hypoxia/reoxygenation model)
• Expression was assessed at both protein level (western blotting) and messenger RNA level (quantitative reverse transcription PCR)
• The effect was dose-dependent across both experimental systems
• H9c2 cardiomyocytes were used as the in vitro hypoxia/reoxygenation model system

YXDW modulated oxidative stress markers in the context of myocardial I/R injury.

• Superoxide dismutase (SOD) was measured as a biochemical marker of oxidative stress response
• Oxidative stress was identified as one of three core pathological pathways (alongside inflammation and apoptosis) modulated by YXDW
• Assessment was conducted using enzyme-linked immunosorbent assay (ELISA) among other biochemical methods