Objective:

2

To investigate the underlying mechanism of Ling-Gui-Zhu-Gan Decoction (LGZGD) in reducing hyperinflammatory responses to provide cardioprotective effects.

Methods:

2

The main chemical components of LGZGD were identified using ultra-high performance liquid chromatography.

In vivo

the rats were randomly divided into 4 groups using simple randomization: the sham group

the model group

the LGZGD (4.2 g/kg) group

and captopril (4.375 mg/kg) group

6 in each group. A rat model of ventricular remodeling (VR) after acute myocardial infarction (AMI) was established by ligation of the left anterior descending coronary artery. After 4 weeks of treatment

cardiac function was evaluated by echocardiography

and histopathological changes were examined using HE and Masson stainings. Serum levels of cardiac enzymes and oxidative stress markers were measured with microplate assays. Reactive oxygen species (ROS) were visualized by fluorescence staining

while protein and mRNA levels were analyzed by Western blot and RT-qPCR. Levels of proinflammatory cytokines and trimethylamine N-oxide (TMAO) were quantified through ELISA.

In vitro

H9c2 cells were exposed to 400 μmol/L TMAO

and oxidative stress markers were measured by microplate assay. ROS levels were visualized using fluorescence staining

and gene and protein expression were analyzed by RT-PCR and Western blot. Proinflammatory cytokines and TMAO levels were further evaluated by ELISA.

Results:

2

LGZGD was found to contain liquiritin

isoliquiritin

coumarin

liquiritigenin

cinnamic acid

kaempferol

cinna

maldehyde

glycyrrhizic acid

and atractylenolide Ⅲ.

In vivo

LGZGD improved cardiac function

reduced myocardial pathology

lowered serum cardiac enzymes and TMAO

decreased oxidative stress

regulated the expression of genes and proteins within the ROS/thioredoxin-interacting protein (TXNIP)/NACHT

LRR

and PYD domains-containing protein 3 (NLRP3) pathway

and suppressed the secretion of IL-1β and IL-18 (

P

<

0.01).

In vitro

analysis showed that LGZGD significantly decreased markers of myocardial injury

alleviated oxidative stress

and inhibited the secretion of IL-1β and IL-18 in TMAO-stimulated H9c2 cells (

P

<

0.01).

Conclusion:

2

LGZGD ameliorates TMAO-induced myocardial injury by modulating the ROS/TXNIP/NLRP3 signaling pathway.