Purpose: Combined with RNA sequencing and experimental verification methods to investigate KXD's potential role and underlying mechanisms in cTnTR141W transgenic mice and H9c2 cells.
Methods: UPLC-Q-TOF-MS analysis was employed to identify the absorbed constituents from KXD in mice blood. cTnTR141W transgenic mice were given with KXD via gavage for 4 weeks. Cardiac remodeling was evaluated by echocardiography, heart weight index, H&E staining, Masson staining, and WGA staining. The potential therapeutic target of KXD was identified through RNA sequencing. H9c2 cells were stimulated with DOX and KXD in vitro. Tunel staining, Immunofluorescence, Transmission electron microscopy, qRT-PCR, and Western blot were used to evaluate cardiomyocyte apoptosis and gap junction remodeling.
Results: 11 compounds from KXD were identified in the blood. KXD improved cardiac dysfunction and alleviated interstitial fibrosis and cardiomyocyte hypertrophy in cTnTR141W transgenic mice. The mechanistic investigations of RNA sequencing revealed that regulating apoptosis and gap junctions may be potential processes for KXD. KXD inhibited cardiomyocyte apoptosis, and promoted the expression of Connexin 43, a key protein of gap junctions, while also inhibiting the phosphorylation of ASK1 and JNK in cTnTR141W transgenic mice and H9c2 cells.
Conclusion: KXD attenuated left ventricular remodeling and cardiomyocyte apoptosis in cTnTR141W transgenic mice and H9c2 cells, potentially through modulation of the ASK1/JNK/Cx43 signaling pathway. These findings provide valuable insights into the therapeutic potential of KXD in mitigating DCM.
Keywords: ASK1/JNK signaling pathway; Kuoxin Decoction; cardiomyocyte apoptosis; dilated cardiomyopathy; gap junctions.
© 2025 Ma et al.