Objective:

2

To evaluate the effects of Sanren Decoction (SRD) on metabolic dysfunction-associated steatotic liver disease (MASLD) induced by high-fat diet (HFD) based on the protective effect of hepatocyte mitochondrial function.

Methods:

2

Thirty-six male C57BL/6 mice were randomly assigned to 4 groups using stratified sampling based on body weight

including control

HFD

low-dose (10.09 g/kg) and high-dose (20.18 g/kg) SRD groups (

n

=9). MASLD model was induced in mice via a 16-week HFD. Liver histopathology was assessed using haematoxylin and eosin (HE) and Oil red O staining

while hepatic triglycerides (TG)

serum alanine aminotransferase (ALT)

fasting blood glucose (FBG)

and fasting serum insulin levels were measured using commercial kits. Hepaticmetabolic profiling were analyzed and differential metabolite analysis was performed using partial least squares discriminant analysis and Kyoto Encyclopedia of Genes and Genomes pathways. Mitochondrial microstructure was assessed by electron microscopy. The protein expressions of respiratory chain complexes Ⅰ–Ⅴ were determined by Western blot analysis

while the activities of complexes Ⅰ and Ⅱ were measured using commercial

kits.

Results:

2

In the HFD-induced MASLD model

4-week SRD treatment improved hepatic steatosis

inflammation

and hepatocyte ballooning (

P

<

0.05). High-dose SRD treatment significantly reduced hepatic TG

ALT levels

and improved insulin sensitivity (both

P

<

0.05). Low-dose SRD significantly reduced hepatic TG and FBG (

P

<

0.05). Metabolomic analysis showed that differential liver metabolites were enriched in tricarboxylic acid cycle (TAC) pathway in mice of high-dose SRD and HFD groups. Electron microscopy showed that high-dose SRD improved mitochondrial morphology and enhanced adenosine triphosphate production and fatty acid oxidation activity (both

P

<

0.05). Additionally

high-dose SRD significantly decreased the contents of hydrogen peroxide and malondialdehyde in liver tissue and increased the content of superoxide dismutase (both

P

<

0.05). Treatment with high-dose SRD resulted in a significant increase in both protein expression and activity of mitochondrial complex Ⅱ. Additionally

high-dose SRD enhanced the protein expression of mitochondrial complex Ⅰ (both

P

<

0.05).

Conclusion:

2

SRD exhibited hepatoprotective effects in MASLD

improving liver morphology

metabolism

and mitochondrial function

suggesting its potential as a therapeutic strategy for MASLD.