Kuoxin Decoction Alleviated Left Ventricular Remodeling by Inhibiting Cardiomyocyte Apoptosis Through ASK1/JNK/Cx43
Mengjiao Ma1, Hua Fan1, Yidan Dong1,2
1Department of Cardiology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, People's Republic of China.
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.
Insights
KXD treatment improved cardiac function and reduced fibrosis in transgenic mice with cardiac troponin T mutations. It works by inhibiting cardiomyocyte apoptosis and modulating the ASK1/JNK/Cx43 pathway, offering potential for treating dilated cardiomyopathy.
Area of Science:
- Cardiovascular Research
- Pharmacology
- Molecular Biology
Background:
- Dilated cardiomyopathy (DCM) is a significant cause of heart failure.
- Genetic mutations, such as in cardiac troponin T (cTnTR141W), can lead to DCM.
- Investigating novel therapeutic compounds for DCM is crucial.
Purpose of the Study:
- To investigate the therapeutic potential of KXD in a mouse model of DCM (cTnTR141W transgenic mice) and in vitro (H9c2 cells).
- To elucidate the underlying molecular mechanisms of KXD's action, focusing on apoptosis and gap junction remodeling.
Main Methods:
- KXD administration via oral gavage to cTnTR141W transgenic mice for 4 weeks.
- Assessment of cardiac remodeling using echocardiography, heart weight index, and histological staining (H&E, Masson, WGA).
- Identification of KXD constituents in blood using UPLC-Q-TOF-MS.
- RNA sequencing to identify therapeutic targets.
- In vitro studies using H9c2 cells treated with DOX and KXD, assessed by Tunel staining, immunofluorescence, TEM, qRT-PCR, and Western blot.
Main Results:
- Eleven KXD compounds were identified in mouse blood.
- KXD treatment significantly improved cardiac dysfunction, reduced interstitial fibrosis, and alleviated cardiomyocyte hypertrophy in cTnTR141W mice.
- Mechanistic studies revealed KXD inhibits cardiomyocyte apoptosis and promotes Connexin 43 (Cx43) expression.
- KXD suppressed the phosphorylation of ASK1 and JNK signaling pathway proteins.
Conclusions:
- KXD effectively attenuated left ventricular remodeling and cardiomyocyte apoptosis in both in vivo and in vitro models.
- The therapeutic effects of KXD are potentially mediated by the modulation of the ASK1/JNK/Cx43 signaling pathway.
- KXD demonstrates significant therapeutic promise for mitigating DCM.
