Objective:To observe the effect of Xiaoyaosan on the activity of the Toll-like receptor-4(TLR-4)/TIR-domain-containing adapter-inducing interferon-β(TRIF) signal transduction pathway in liver cells of rats of non-alcoholic fatty hepatitis (NASH) with liver-Qi stagnation and spleen deficiency syndrome. Method:Thirty-two SPF grade SD male rats were selected and randomly divided into normal control group

model group

Xiaoyaosan treatment group and glycine treatment group (n=8). Except for the rats in normal control group

which were fed routinely with normal die

the NASH with the liver-Qi stagnation and the spleen deficiency syndrome experimental model were established in the rats of the other 3 groups with the ‘high-glucose and high-fat feed+irregular eating+chronic bondage stimulation’ method for 14 weeks. And then

normal saline

Xiaoyaosan solution (1 g·mL-1) and glycine solution (2.5 g·mL-1) were given by gavage for 4 weeks at the dosage of 2 mL·kg-1·d-1

3.24 mL·kg-1·d-1 and 2 mL·kg-1·d-1 in model group

Xiaoyaosan treatment group and glycine treatment group

respectively. The syndrome scores of liver-Qi stagnation and spleen deficiency were calculated

and the urine D-xylose excretion rate was detected in all of rats before being put to death

and the liver and brain tissues were extracted under aseptic and low temperature conditions. The content of norepinephrine (NE) and 5-serotonin (5-HT) in brain

the degree of steatosis and inflammatory cell infiltration and activity of TLR-4/TRIF signal transduction pathways in liver were detected with enzyme linked immunosorbent assay(ELISA)

htoxylin eosin(HE) staining and Real-time PCR

western blot technique. Result:Compared with the normal control group

increased syndrome scores of liver-Qi stagnation and spleen deficiency syndrome

declined urine D-xylose excretion rate and content of 5-HT and NE in brain

lots of fat vacuoles in hepatocyte cytoplasm and scattered spotty necrosis and inflammatory cells infiltration in hepatic lobular

increased TLR-4

TRIF mRNA and protein expressions in liver could be found in rats of model group (P<0.05). Compared with the model group

decreased syndrome scores of liver-Qi stagnation and spleen deficiency syndrome

elevated urine D-xylose excretion rate and content of 5-HT and NE in brain

alleviated steatosis and inflammatory cell infiltration

declined TLR-4

TRIF mRNA and protein expressions in liver could be found in rats of Xiaoyaosan treatment group and glycine treatment group (P<0.05). Conclusion:Xiaoyaosan could play the role in the treatment of NASH with liver-Qi stagnation and spleen deficiency syndrome by dispersing stagnated liver-qi

strengthening spleen and inhibiting activity of TLR-4/TRIF signal transduction pathway.