Pinggan Yishen formula alleviates cardiovascular remodeling in SHR while modulating vascular MLCK and cardiac Tsc22D4

Ping-Gan-Yi-Shen formula (PGYS) is mainly used for patients with severe hypertension accompanied by dizziness and headache, and has shown good clinical efficacy. In this study, liquid chromatography-tandem mass spectrometry (LC-MS/MS), targeted network pharmacology, multi-omics analysis, as well as molecular docking, and in vivo experiments were employed to systematically analyze the blood-absorbed components of PGYS, predict its active ingredients, and validate its mechanisms of action. A total of 8 blood-absorbed components were identified in the serum of PGYS-treated rats. Through network pharmacology analysis, myosin light chain kinase (MLCK) was screened as a key target. In vivo experiments showed that PGYS significantly reduced blood pressure, improved cardiac function, and attenuated vascular and cardiac fibrosis. Multi-omics analysis revealed that PGYS downregulated the expression of Tsc22 domain family member 4 (Tsc22D4) in cardiac tissue, thereby improving myocardial insulin sensitivity and reducing lipid accumulation. Molecular docking confirmed that caffeic acid and cellobiose, stably bind to MLCK and Tsc22D4 proteins. Furthermore, in vivo experiments demonstrated that PGYS alleviated cardiovascular remodeling, potentially by modulating vascular MLCK and cardiac Tsc22D4, and these regulatory effects were associated with decreased levels of TGF-β1 and ApoE, enhanced inositol metabolism, and reduced insulin resistance. These findings indicate that PGYS exerts antihypertensive and cardioprotective effects putatively by regulating vascular MLCK and cardiac Tsc22D4 to mitigate cardiovascular remodeling.