Background: Shenfu Injection (SFI) has been widely utilized in the treatment of cardiovascular diseases in China for over 3 decades. In recent years, the therapeutic role of SFI in cardiovascular diseases has garnered significant attention. This study aims to elucidate the effects and possible mechanism of SFI using in vitro experiments and network pharmacology analysis.

Methods: Hypertrophy was induced by Angiotensin II (Ang II) treatment of HL-1 and isoprenaline (ISO) treatment of AC16. First, cell viability was detected upon different concentrations of SFI to screen the safe dosage of SFI. The effect of SFI on cell functions was evaluated by cell size, atrial natriuretic peptide/brain type natriuretic peptide (ANP/BNP) levels, mitochondrial morphology, and Ser616/Ser579 or Ser637/Ser600 levels. The potential targets of SFI against cardiomyocyte hypertrophy were predicted by network pharmacology, and then experimentally validated by qRT-PCR.

Results: Shenfu Injection treatment significantly reduced cell size, ANP expression, BNP expression, and mitochondrial injury compared to Ang and ISO groups (P < .001). Western blot analysis showed that SFI decreased the expression of Dynamin-related protein 1 (Drp1) phosphorylation at Ser616/Ser579 and increased the expression of Drp1 phosphorylation at Ser637/Ser600 in Ang- and ISO-induced cells. Network pharmacology analysis and cell experiments showed SFI decreased dopamine receptor D2 (DRD2) and increased protein kinase A levels in induced cardiomyocytes.

Conclusion: In summary, the findings demonstrate that SFI may alleviate cardiomyocyte hypertrophy by targeting DRD2 and inhibiting mitochondrial fission.

#Yuhan Zhang and Su Zhang contributed equally to this work